WO2011051327A2 - Small antibody-like single chain proteins - Google Patents

Abstract

The present invention provides a "ASCP-body" that comprises a bottomside epitope binding domain ("BEBD") comprising four Bottomside Binding Regions (BBR1, BBR2, BBR3, and BBR4). This epitope binding domain is in addition to the topside epitope binding domain (TEBD) formed by the three Complementarity Determining Regions (CDR1, CDR2, and CDR3). The invention encompasses mono-specific, bi-specific and multi-specific, and mono-valent, bi-valent and multi-valent ASCP-bodies. The invention also encompasses fusion proteins in which one or more ASCP-bodies with a BEBD is conjugated to one or more ASCP-bodies with a BEBD and, optionally, a TEBD; to one or more ASCP- bodies with a TEBD and, optionally, a BEBD; or to one or more non-ASCP-body entity(ies). The various ASCP-bodies in the bi-specific and multi-specific ASCP-bodies and fusion proteins may all bind to the same epitopes or antigens or may bind to different epitopes or antigens.

Description

SMALL ANTIBODY-LIKE SINGLE CHAIN PROTEINS

CROSS-REFERENCE TO RELATED APPLICATIONS

[001] This application claims benefit of U.S. Application Serial Number 61/256,638 filed

October 30, 2009, the disclosures of which are hereby incorporated by reference.

BACKGROUND OF THE INVENTION

[002] Molecules capable of specific binding to a desired target epitope are of enormous importance as both therapeutics and medical diagnostic tools. A well known example of this class of molecules is the monoclonal antibody. A monoclonal antibody comprises an epitope binding domain comprising three Complementarity Determining Regions ("CDRs"). Antibodies can be selected that bind specifically and with high affinity to almost any structural epitope. As a result, antibodies are used routinely as research tools and as FDA approved therapeutics such that the worldwide market for therapeutic and diagnostic monoclonal antibodies is currently worth approximately $30 billion.

[003] However, monoclonal antibodies have a number of shortcomings. For example, classical antibodies are large and complex molecules. They have a heterotetrameric structure comprising two light chains and two heavy chains connected together by both inter and intra disulphide linkages. This structural complexity precludes easy expression of antibodies in simple prokaryotic systems and requires that antibodies are produced in more elaborate (and expensive) mammalian cell systems. The large size of antibodies also limits their therapeutic effectiveness since they are often unable to efficiently penetrate certain tissue spaces. Therapeutic antibodies, because they possess an Fc region, occasionally trigger undesired effector cell function and/or clotting cascades. In addition, generating bi-specific or multi-specific antibodies often involves difficult and complex procedures (See e.g., Josefina et al, (1997), Nature Biotechnology, 15: 159-163; and Wu et al. (2007) Nature Biotechnology, 25: 1290-1297).

[004] Accordingly there is a need in the art for alternative binding molecules capable of specific binding to a desired target with high affinity and specificity. A need also exists for a simple method to generate bi-specific or multi-specific binding molecules.

SUMMARY OF THE INVENTION

[005] The present invention provides a antibody-like single chain proteins ("ASCPs") or

("ASCP-bodies"), such as a NANOBODY^® that comprises a Bottomside Epitope Binding Domain. A NANOBODY^® usually comprises an epitope binding domain (the Topside Epitope Binding Domain, or "TEBD") comprising three complementarity determining regions referred to herein as CDR1, CDR2 and CDR3. The present invention is based on the unexpected and surprising discovery that a variant of a NANOBODY^® may further comprise one or more additional epitope binding domains, the Bottomside Epitope Binding Domain (or "BEBD"). The BEBD in the ASCP- body of the invention comprises four Bottomside Binding Regions ("BBRs"), namely BBR1, BBR2, BBR3, and BBR4. The ASCP-body of the invention would have at least one BEBD, while optionally containing one or more additional BEBDs or TEBDs in the same single domain.

[006] The invention further provides fusion proteins in which one or more ASCP-bodies having a BEBD is conjugated operably linked to one or more ASCP-bodies with a TEBD. The various epitope binding domains in the BEBDs and/or TEBDs of the ASCP fusion protein may all bind to the same antigen. Alternatively, the various epitope binding domains of the ASCP fusion protein may bind to different antigens, thus forming a bi-specific or multiply-specific fusion protein.

[007] The invention also relates to nucleic acids encoding the ASCPs; to methods for preparing the ASCP-bodies; to host cells expressing or capable of expressing ASCP-bodies; to compositions, and in particular to pharmaceutical compositions, that comprise the ASCP-bodies, nucleic acids and/or host cells; and to uses of such ASCP-bodies, nucleic acids, host cells and/or compositions, in particular for prophylactic, therapeutic or diagnostic purposes, such as the prophylactic, therapeutic or diagnostic purposes mentioned herein.

[008] The invention also relates to functional fragments, BBR1 , BBR2, BBR3, and BBR4 sequences, BEBD sequences, parts, fragments, analogs, mutants, variants, alleles and/or derivatives of the ASCP-bodies and polypeptides of the invention, and/or to use proteins or polypeptides comprising or essentially consisting of the same, as long as these are suitable for the uses envisaged herein. Such parts, fragments, analogs, mutants, variants, alleles, derivatives, proteins and/or polypeptides will be described in the further description herein.

[009] Accordingly, the invention pertains to at least the following various non-limiting embodiments.

[0010] In one embodiment, the present invention pertains to: a ASCP-body comprising a bottomside epitope binding domain comprising Bottomside Binding Region ("BBR") 1 , BBR 2, BBR3, and BBR4.

[001 1] In one embodiment of the ASCP-body of the present invention, the bottomside epitope binding domain binds to a half-life extender.

[0012] In one embodiment of the ASCP-body of the present invention, the half-life extender is human serum albumin.

[0013] In one embodiment, the present invention pertains to: a pharmaceutical composition of this ASCP-body.

[0014] In one embodiment, the present invention pertains to: a ASCP-body comprising a bottomside epitope binding domain comprising BBR (Bottomside Binding Region) 1 , BBR 2 and BBR3, and also comprising a topside epitope binding domain comprising CDR (complementarity determining region) 1 , CDR2 and CDR3.

[0015] In one embodiment of the ASCP-body of the present invention, the topside epitope binding domain binds to the same antigen as the epitope bound by the bottomside epitope binding domain.

[0016] In one embodiment of the ASCP-body of the present invention, the topside epitope binding domain binds to a different antigen as the epitope bound by the bottomside epitope binding domain.

[0017] In one embodiment of the ASCP-body of the present invention, the topside epitope binding domain binds to a half- life extender.

[0018] In one embodiment of the ASCP-body of the present invention, the half-life extender is human serum albumin.

[0019] In one embodiment of the ASCP-body of the present invention, the bottomside epitope binding domain binds to a half-life extender.

[0020] In one embodiment of the ASCP-body of the present invention, the half-life extender is human serum albumin.

[0021 ] In one embodiment, the present invention pertains to: a pharmaceutical composition of this ASCP-body.

[0022] In one embodiment, the present invention pertains to: a composition comprising a

ASCP-body comprising a bottomside epitope binding domain comprising BBR (Bottomside Binding Region) 1, BBR 2 and BBR3, and also comprising a topside epitope binding domain comprising CDR (complementarity determining region) 1, CDR2 and CDR3, wherein the ASCP-body is conjugated to one or more additional ASCP-bodies comprising a bottomside epitope binding domain comprising BBR (Bottomside Binding Region) 1 , BBR 2 and BBR3, and also comprising a topside epitope binding domain comprising CDR (complementarity determining region) 1, CDR2 and CDR3.

[0023] In one embodiment of this composition of the present invention, the topside epitope binding domain of the first ASCP-body and the topside epitope binding domain of the one or more additional ASCP-bodies bind to the same antigen.

[0024] In one embodiment of this composition of the present invention, the topside epitope binding domain of the first ASCP-body and the topside epitope binding domain of the one or more additional ASCP-bodies bind to different antigens.

[0025] In one embodiment of this composition of the present invention, at least one of the topside or bottom side epitope binding domains binds to a half-life extender.

[0026] In one embodiment of this composition of the present invention, the half-life extender is human serum albumin. [0027] In one embodiment, the present invention pertains to: a pharmaceutical composition of this composition.

[0028] In one embodiment, the present invention pertains to: a composition comprising one or more first ASCP-bodies of claim 1, conjugated to one or more second ASCP-bodies, wherein the one or more second ASCP-bodies comprise a topside epitope binding domain comprising CDR (complementarity determining region) I , CDR2 and CDR3.

[0029] In one embodiment of this composition of the present invention, the bottomside epitope binding domain of the one or more first ASCP-bodies binds to the same antigen as the topside epitope binding domain of the one or more second ASCP-bodies.

[0030] In one embodiment of this composition of the present invention, the bottomside epitope binding domain of the one or more first ASCP-bodies binds to a different antigen than the topside epitope binding domain of the one or more second ASCP-bodies.

[0031 ] In one embodiment of this composition of the present invention, at least one of the topside or bottom side epitope binding domains binds to a half-life extender.

[0032] In one embodiment of this composition of the present invention, the half-life extender is human serum albumin.

[0033] In one embodiment, the present invention pertains to: a pharmaceutical composition of this composition.

[0034] In one embodiment, the present invention pertains to: a composition comprising a first ASCP-body comprising a bottomside epitope binding domain comprising BBR (Bottomside Binding Region) 1 , BBR 2, BBR3, and BBR4, conjugated to one or more additional ASCP-bodies comprising a bottomside epitope binding domain comprising BBR (Bottomside Binding Region) 1 , BBR 2, BBR3, and BBR4.

[0035] In one embodiment of this composition of the present invention, the bottomside epitope binding domain of the first ASCP-body and the bottomside epitope binding domains of the one or more additional ASCP-bodies bind to the same antigen.

[0036] In one embodiment of this composition of the present invention, the bottomside epitope binding domain of the first ASCP-body and the bottomside epitope binding domains of the one or more additional ASCP-bodies bind to different antigens.

[0037] In one embodiment of this composition of the present invention, at least one bottomside epitope binding domain binds to a half-life extender.

[0038] In one embodiment of this composition of the present invention, the half-life extender is human serum albumin.

[0039] In one embodiment, the present invention pertains to: a pharmaceutical composition of this composition. [0040] In one embodiment, the present invention pertains to: an ASCP-body comprising a bottomside epitope binding domain comprising BBR (Bottomside Binding Region) 1, BBR 2, BBR3, and BBR4, wherein the ASCP-body is conjugated to a non-ASCP-body entity.

[0041] In one embodiment of this composition of the present invention, the non-ASCP- body entity is one or more proteins, peptides, carbohydrates and/or lipids.

[0042] In one embodiment, the present invention pertains to: a composition comprising a

ASCP-body comprising a bottomside epitope binding domain comprising BBR (Bottomside Binding Region) 1, BBR 2, BBR3, and BBR4, further comprising a CHI, CH2, and/or CH3. In this embodiment, the ASCP-body resides in the context of a SCALP or SmALP, as defined herein.

[0043] In one embodiment, the present invention pertains to: a library of ASCP-bodies comprising a bottomside epitope binding domain comprising BBR (Bottomside Binding Region) 1 , BBR 2, BBR3, and BBR4.

[0044] In one embodiment, the present invention pertains to: a method of creating a library of ASCP-bodies comprising a bottomside epitope binding domain comprising BBR (Bottomside Binding Region) 1, BBR 2, BBR3, and BBR4, the method comprising the steps of providing a VHH domain directed to a desired antigen:

(a) by a method comprising the steps of:

(i) immunizing a mammal belonging to the Camel idae with the desired antigen or a fragment thereof, so as to raise an immune response and/or antibodies (particularly heavy chain antibodies) against the antigen;

(ii) obtaining a biological sample from the immunized mammal, wherein the sample comprises heavy chain antibody sequences and/or V_HH sequences directed against the antigen; and

(iii) obtaining the BBR sequences from the heavy chain antibody sequences and/or V_HH sequences; or

(b) by a method comprising the steps of

(i) screening a library comprising heavy chain antibody sequences and/or V_HH sequences for sequences directed to the antigen;

(ii) obtaining the heavy chain and/or V_M sequences from the library; and

(iii) obtaining the BBR sequences from the heavy chain and/or V_HH sequences.

[0045] In one embodiment of this method of the present invention, the method further comprises the step of (b) mutagenizing the VHH domain.

[0046] Other features and advantages of the invention will be apparent from the following detailed description and claims. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] Figure 1 A shows a schematic diagram of the beta pleated sheets, alpha helix and loops that comprise a stereotypical ASCP-body; the amino (N) and carboxy (C) terminus of the protein are indicated, where numbers represent the corresponding amino acid residues within the single chain ASCP-body. Figure 1 B shows an exemplary sequence of a representative ASCP-body, where the numbered and bolded residues labeled in library 1 (top side library) indicate the CDR1 , CDR2 and CDR3 residues of a representative sequence (SEQ ID NO: 1 ), and the numbered and bolded residues labeled in library 2 (bottom side library) indicate the BBR1. BBR2. BBR3 and BBR4 residues in the same sequence. The residue numbers in Figure 1 A and Figure IB are meant to correspond.

[0048] Figure 2A diagrams a monoclonal antibody and various SCALPS (single-chain antibody-like proteins) and SmALPs (small antibody-like proteins). Indicated are variable regions of the light (VL or V_L) and heavy chains (VH or V_H); the first constant region (CHI) of the heavy chain; the Fc portion comprising the constant heavy chain 2 (CH2) and constant heavy chain 3 (CH3) regions, and various SCALPS and SmALPs, including Fab, Fv, and ScFv fragments. Figure 2A diagrams a camelid single chain antibody, including the Fc portion and a V_HH fragment.

DETAILED DESCRIPTION OF THE INVENTION

[0049] In order to provide a clear understanding of the specification and claims, the following definitions are conveniently provided below.

[0050] As "ASCP-body" refers to a type of single-chain antibody-like protein (SCALP) comprising a variable region (V_HH) of a heavy chain of a camelid antibody. In a particular embodiment of the invention, the ASCP-body comprises two epitope binding domains - namely one on the topside and one on the bottomside. One epitope binding domain, the bottom epitope binding domain (BEBD) is comprised of four sequences, the bottom binding regions BBR1 , BBR2, BBR3, and BBR4 . The other epitope binding domain is the top epitope binding domain (TEBD), which is comprised of the complementarity determining regions CDR1, CDR2 and CDR3. The term

"complementarity determining region (CDR)" refers to a hypervariable loop from an antibody variable domain or from a T-cell receptor. The position of CDRs within a antibody variable region have been precisely defined (see, Kabat, E.A., et al. Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, N1H Publication No. 91- 3242, 1991, and Chothia, C. et al, J. Mol. Biol. 196:901-917, 1987, which are incorporated herein by reference). The specific residues for BBR1 , BBR2, BBR3, BBR4, CDR1 , CDR2 and CDR3 are provided in Figure 1 and Table 1 , plus in representative consensus sequences provided herein.

[0051] Around and between the BBRs and CDRs of the various BEBD and TEBD sequences are framework (FR) sequences. [0052] The term "SCALP" refers to a Single-Chain Antibody-like Protein. The grouping

SCALP includes an isolated heavy chain of an antibody, or fragment, thereof; or one variable region fused to another. Thus, the group of SCALPS includes V_HH (ASCP-bodies); and VHH plus any one or more constant region of the heavy chain (e.g., CH 1 , CH2, and/or CH3). Thus, a SCALP is exemplified by a VHH; a V_HH + CHI ; a VHH + CH2; a VHH + CH3; a V_HH + CH 1 + CH2; a V_HH + CH 1 + CH2 + CH3; a VHH + CH2 + CH3 ; a V_HH + CH 1 + CH3; etc. SCALPs include single domain antibodies. The term "single domain antibodies" refers to any naturally-occurring single variable domain and corresponding engineered binding fragments, including human domain antibodies as described by Domantis (Domantis / GSK (Cambridge, UK) or camelid ASCP-bodies as defined hereafter. The term "camelid ASCP-body" refers to a region of camelid antibody which is the small single variable domain devoid of light chain and that can be obtained by genetic engineering to yield a small protein having high affinity for a target, resulting in a low molecular weight antibody-derived protein. For the "camelid Nanobody" see WO07042289 and U.S. patent number 5,759,808 issued June 2, 1998; see also Stijlemans, B. et al., 2004. Engineered libraries of camelid antibodies and antibody fragments are commercially available, for example, from Ablynx, Ghent, Belgium. As with other antibodies of non-human origin, an amino acid sequence of a camelid antibody can be altered recombinantly to obtain a sequence that more closely resembles a human sequence, i.e., the ASCP-body can be "humanized". Thus the natural low antigenicity of camelid antibodies to humans can be further reduced. The group of SCLAPs also includes single chain antibodies. The term "single chain antibody" refers to an antigen binding portion of a light chain variable region and an antigen binding portion of a heavy chain variable region, joined, using recombinant methods, by a synthetic linker that enables them to be made as a single protein chain in which the VL and VH regions pair to form monovalent molecules (known as single chain Fv (scFv); see e.g., Bird et al. (1988) Science 242:423-426; and Huston et al ( 1988) Proc. Natl. Acad. Sci. U.S.A 85:5879-5883).

[0053] By "SmALP" is meant any Small Antibody-like Protein. SmALPs include Fab fragments, which comprise the V_H, CH, V_L and V_H portions; Fv, which comprise the V_H and V_L portions; ScFv, which comprise a V_H and V_L portion fused to each other; dAbs; di-ScFv, which comprise two ScFv fragments; and Fcabs. These and other SmALPs are described in Classical and Heavy Chain Antibodies, from Muyldermans, S. (2001 ), J Biotechnol. 74, 277-302, and are well- known in the art. dAbs (domain antibodies) correspond to the variable regions of either the heavy (V_H) or light (V_L) chains of human antibodies and have a molecular weight of approximately 13 kDa, or less than one-tenth the size of a full antibody. dAbs are available from Domantis Limited, a wholly owned subsidiary of GlaxoSmithKline. An Fcab (antigen-binding Fc) is a compressed antibody, comprising a CH2 and CH3 domain, with two identical antigen binding sites engineered into the CH3 domains, as available from f-star Biotechnologische Forschungs- und Entwicklungsges, M.b.H. A ASCP-body of the present invention can be fused to a CHI and/or CH2 and/or CH3, and/or V_L region. The ASCP-body can thus be within the context of a SCALP or SmALP.

[0054] The term "mono-specific" as used herein refers to an ASCP-body that binds to only one target molecule of interest. For example, the BEBD can bind to a target of interest, but the TEBD does not. in another embodiment of a mono-specific ASCP-body, both the BEBD and TEBD bind to the same target. In various embodiments of a mono-specific ASCP-body, the BEBD and TEBD can bind to the same or different epitopes within the same target. It is to be understood that not all the loops from the top or bottom region need to be used for binding the target molecule; e.g., only the BBR1 and BBR3 but not the BBR2, or only the BBR2 and BBR3 but not BBR1 may be involved in binding.

[0055] In addition, a chain or conjugate of two or more mono-specific ASCP-bodies can be formed; if they all bind to the same target, a mono-specific composition with more than a valence of 1 is formed. For example, a mono-specific ASCP-body can have a BEBD that binds to one target. Two such ASCP-bodies linked would form a mono-specific, bivalent entity. Three or more such ASCP-bodies linked or fused together would form a mono-specific, multivalent entity.

[0056] The term "bi-specific" as used herein refers to a ASCP-body or conjugate of two or more ASCP-bodies which bind to two distinct targets of interest. In one embodiment of a single bi- specific ASCP-body, the TEBD binds to one target, while the BEBD binds to a different target. In another embodiment, the "bi-specific" composition comprises a fusion of two or more ASCP-bodies. In this embodiment, at least one binding domain (e.g., either a BEBD or TEBD) binds to one target, while another binding domain (e.g., either a BEBD or a TEBD, preferably on a different ASCP- body) binds to a second target. For example, a bi-specific composition can be a ASCP-body with a BEBD that binds to one target, fused to a second ASCP-body with a TEBD that binds to another target. In another embodiment, the bi-specific composition can be a ASCP-body with a BEBD that binds to one target, fused to a second ASCP-body with a BEBD that binds to another target.

[0057] The term "multi-specific" refers to a composition which binds to more than one target. For example, various mono-specific ASCP-bodies (each of which bind to a single, different target) can be conjugated, forming a composition capable of binding to more than two targets simultaneously. A multi-specific composition also includes, for example, one ASCP-body with a BEBD that binds one target and a TEBD that binds to a second target, fused to a second ASCP-body with a BEBD that binds to a third target. The TEBD of the second ASCP-body can optionally target a fourth target.

[0058] The term "target" refers to an antigen or epitope recognized by ASCP-body of the invention. Targets include, but are not limited to, epitopes present on proteins, peptides, carbohydrates, and/or lipids. Targets include, inter alia, half-life extenders (e.g., human serum albumin, PEG, or transferring), cellular receptors, cellular receptor ligands, bacterial or viral proteins or other components, or proteins or other markers or components associated with or involved in human disease, e.g., autoimmune disease, cancer or an infectious disease.

[0059] The term "conjugate" refers to an ASCP-body chemically or genetically linked to one or ASCP-bodies, or non-ASCP-body entity (non-ASCP-body), e.g., another protein, peptide, carbohydrate and/or lipid, or one or more of these. A ASCP-body in the conjugate can be conjugated to the other moiety at the amino-terminus (N-terminus), carboxy-terminus (C-terminus), or any other portion of the ASCP-body. For example, the N-terminus of one ASCP-body can be conjugated to the C-terminus of another ASCP-body or non-ASCP-body. In another non-limiting example, the two C-termini of two ASCP-bodies can be conjugated together.

[0060] The term "non-ASCP-body" refers to a biological or chemical entity that imparts additional ftinctionality to a molecule to which it is attached. In a particular embodiment, the non- ASCP-body is a polypeptide, e.g., human serum albumin (HSA), or a chemical entity, e.g., polyethylene glycol (PEG), fused to the ASCP-body, which increases the half-life of the ASCP-body in vivo. Non-limiting examples of non-ASCP-body entities that can be conjugated to a ASCP-body include: toxins, other portions of a monoclonal antibody (e.g., CHI, CH2, CH3, C_L, and/or V_L), epitope tags (e.g., 6xHis), Fc regions, Human Serum Albumin (HSA) (or portions thereof), polyethylene glycol (PEG) and/or polypeptides which bind to the aforementioned proteins or other serum proteins with increased half-life, such as, e.g., transferrin. The term "polyethylene glycol" or "PEG" refers to a polyalkylene glycol compound or a derivative thereof, with or without coupling agents or derivatization with coupling or activating moieties.

[0061] The term "non-natural amino acid residue" refers to an amino acid residue that is not present in the naturally occurring (wild-type) protein. Such non-natural amino acid residues can be introduced by substitution of naturally occurring amino acids, and/or by insertion of non-natural amino acids into the naturally occurring amino acid sequence. The non-natural amino acid residue also can be incorporated such that a desired functionality is imparted to the ASCP-body, for example, the ability to link a functional moiety (e.g., PEG).

[0062] The term "polyethylene glycol" or "PEG" refers to a polyalkylene glycol compound or a derivative thereof, with or without coupling agents or derivatization with coupling or activating moieties.

[0063] The term "specific binding'" or "specifically binds to" refers to the ability of an

ASCP-body to bind to a target with an affinity of at least 1 x 10^"6 M, and/or bind to a target with an affinity that is at least two-fold, (preferably at least 10 fold), greater than its affinity for a nonspecific antigen at room temperature under standard physiological salt and pH conditions, e.g., as measured by surface plasmon resonance.

[0064] The term "half-life extender" refers to a chemical entity which, when conjugated to a

ASCP-body, extends the half-life of that ASCP-body in the bloodstream. Examples of half-life extenders include human serum albumin, PEG (polyethylene glycol), and other extenders known in the art. The non-ASCP-body moiety of the conjugate can be conjugated to either the amino terminus (N-terminus) or carboxy terminus (C-terminus) of the ASCP-body, or anywhere in between.

[0065] ASCP-bodies comprising a bottomside epitope binding domain (BEBD)

[0066] The present invention relates to a polypeptide construct comprising one or more

ASCP-bodies. The ASCP-bodies comprise a Bottomside Epitope Binding Domain or BEBD. The (BEBD) comprises four Bottomside Binding Regions (BBR1, BBR2, BBR3, and BBR4 ).

[0067] ASCP-bodies (a.k.a. single domain antibodies or V_HH antibodies) are antibody fragments consisting of a single monomeric variable antibody domain. They are derived, for example, from heavy chain antibodies from camelids, which consist only of two antibody heavy chains, with no light chain. With a molecular weight of only 12-15 kDa, ASCP-bodies are much smaller than monoclonal antibodies (MAbs), e.g., IgG antibodies (150-160 kDa), which have two heavy protein chains and two light chains. ASCP-bodies of the invention would have multiple pharmaceutical applications and have potential for use in treatment of cancer, Alzheimer's Disease, and other diseases.

[0068] Like Nanobodies, ASCP-bodies have advantages over monoclonal antibodies.

Traditional therapeutic monoclonal antibodies must be stored at near freezing temperatures to prevent their destruction. Antibodies are not suited for oral administration because they are digested quickly in the gut, and are not usually useful for treating diseases of the brain because they do not easily permeate the blood-brain barrier. Additionally, therapeutic antibodies are not well suited to target large tumors because they are held to the periphery of solid tumors. Many illnesses are thus unreachable by monoclonals, and patients who use MAb therapies must receive them by injection or infusion at a clinic. Because ASCP-bodies are so much smaller than antibodies and are not hydrophobes (as are standard human antibodies), they would be more resistant to heat and pH, and may retain their activity as they pass through the gastrointestinal tract, raising the prospect of oral ASCP-body pills to treat inflammatory bowel disease, colon cancer, and other disorders of the gut. See Nanobody studies reported by Harmsen et al. (2007) Appl Microbiol. Biotechnoi. 77 (1): 13-22; Hamers-Casterman et al. Nature. 1993, 363(6428):446-8.

[0069] ASCP-bodies are antibodies whose complementary determining regions are part of a single domain polypeptide. Examples include, but are not limited to, heavy chain antibodies, antibodies naturally devoid of light chains, Nanobodies derived from conventional 4-chain antibodies, engineered antibodies and single domain scaffolds other than those derived from antibodies. ASCP-bodies may be any of the art, or any future ASCP-bodies. ASCP-bodies may be derived from any species including, but not limited to mouse, human, camel, llama, goat, rabbit, bovine. According to one aspect of the invention, a ASCP-body as used herein is a modification of a naturally occurring immunoglobulin known as heavy chain antibody devoid of light chains. Such immunoglobulins are disclosed in WO 9404678, and published U.S. Patent Application

20090238829 to Silence et al. for example. For clarity reasons, the variable domain derived from a heavy chain antibody naturally devoid of light chain is known herein as a VHH or ASCP-body to distinguish it from the conventional V_H of four chain immunoglobulins. Compare Fig. 2A and 2B. Such a VHH molecule can be derived from antibodies raised in Camelidae species, for example in llama, camel, dromedary, alpaca and guanaco. Other species besides Camelidae may produce heavy chain antibodies naturally devoid of light chain; such V_HHS are within the scope of the invention.

[0070] The properties of ASCP-bodies, in particular VHHS, compare favourably with those of antibodies derived from sources such as mouse, sheep, goat, rabbit etc. (i.e. traditional antibodies), and humanised derivatives thereof. Traditional antibodies are not stable at room temperature, and have to be refrigerated for preparation and storage, requiring necessary refrigerated laboratory equipment, storage and transport, which contribute towards time and expense. Refrigeration is sometimes not feasible in developing countries. Furthermore, the manufacture or small-scale production of said antibodies is expensive because the mammalian cellular systems necessary for the expression of intact and active antibodies require high levels of support in terms of time and equipment, and yields are very low.

[0071 ] Traditional antibodies have a binding activity which depends upon pH, and hence are unsuitable for use in environments outside the usual physiological pH range such as, for example, in treating gastric bleeding, gastric surgery. Furthermore, traditional antibodies are unstable at low or high pH and hence are not suitable for oral administration. However, it has been demonstrated that VHHS resist harsh conditions, such as extreme pH, denaturing reagents and high temperatures (Ewert S et ai, Biochemistry 2002 Mar. 19; 41(1 1):3628-36), so making them suitable for delivery by oral administration. Finally, traditional antibodies have a binding activity that depends upon temperature, and hence are unsuitable for use in assays or kits performed at temperatures outside biologically active-temperature ranges.

[0072] In contrast, VHHS, and hence ASCP-bodies, are more soluble, meaning they may be stored and/or administered in higher concentrations compared with conventional antibodies. ASCP- bodies of the invention retain binding activity at a pH and temperature outside those of usual physiological ranges, which means they may be useful in situations of extreme pH and temperature which require a modulation of platelet-mediated aggregation, such as in gastric surgery, control of gastric bleeding, assays performed at room temperature etc. The ASCP-bodies also exhibit a prolonged stability at extremes of pH, meaning they would be suitable for delivery by oral administration. ASCP-bodies may be cost-effectively produced through fermentation in convenient recombinant host organisms such as Escherichia coli and yeast; unlike conventional antibodies which require expensive mammalian cell culture facilities, achievable levels of expression for ASCP-bodies are high. Examples of yields of ASCP-bodies are 1 to 10 mg/ml (E. coli) and up to 1 g 1 (yeast). The polypeptides of the present invention also exhibit high binding affinity for a broad range of different antigen types, and ability to bind to epitopes not recognised by conventional antibodies; for example they display long CDR-based loop structures with the potential to penetrate into cavities and exhibit enzyme function inhibition. Furthermore, since binding often occurs through the CDR3 loop only, it is envisaged that peptides derived from CDR3 could be used therapeutically (Desmyter et al., J Biol Chem, 2001, 276: 26285-90). The ASCP-bodies of the invention are also able to retain full binding capacity as fusion protein with an enzyme or toxin.

[0073] Nanobodies generally comprise a single amino acid chain and are known to comprise framework regions (FR) and a single epitope binding domain, comprised of CDR

(complementarity determining region) 1, CDR2, and CDR3. The binding domain consisting of the three CDR sequences has been re-designated the topside epitope binding domain (TEBD).

[0074] The Applicants have discovered that Nanobodies contain, or can be engineered to contain, an additional epitope binding domain, thus engineering an ASCP-body of the invention. This epitope domain is on the bottomside of the ASCP-body and has been designated the bottomside epitope binding domain (BEBD), which consists of four sequences, bottomside binding regions BBR1, BBR2, BBR3, and BBR4.

[0075] The ASCP-bodies of the invention may be engineered to remove the three CDRs of the TEBD and would still have the full epitope binding ability provided by the BEBD. Thus, a single chain of a ASCP-body comprises various intermittent framework sequences (FR); bottomside epitope binding domain sequences which comprise bottomside binding regions BBR1 , BBR2, BBR3, and BBR4 ; and, optionally, topside epitope binding domain sequences (TEBD), comprising complementarity determining regions CDR1, CDR2 and CDR3.

[0076] The ASCP-bodies of the invention may be engineered to include additional single chain antibody fragments, such as CHI , CH2, CH3 regions to generate SCALPS. In some embodiments, the CHI , CH2, CH3 regions are engineered with mutations such that each CHI , CH2, CH3 region can independently bind to a target. These mutations include mutations in which selected amino acid residues in the wildtype sequence of the CHI , CH2, CH3 regions are replaced by randomly chosen amino acids, selective amino acids, as well as insertions or deletions of amino acid residues. The mutations can be made to any region of the CH 1 , CH2, or CH3 domain, such as any solvent exposable region, top region of the CH domain, bottom region of the CH domain, or any combination of the top and bottom region of the CH domain. In one embodiment, the loop regions of CHI , CH2 or CH3 are mutated. These loop regions comprise amino acids 8-20, 24-39, 42-78, 82- 85, 91-103 and 108-1 17. Solvent exposable regions or loop regions of CH domains can readily be identified from X-ray chrystallography libraries of single domain antibodies, available from a umber of databases, e.g., the Brookhaven Database. [0077] In one embodiment, a SCALP contains at least one domain of the variable region and at least one domain of the constant region, e.g., an ASCP with a BEBD, and at least one CH domain selected from the group consisting of CHI , CH2, or CH3 in which the top region, bottom region, or any combination of top and/or bottom regions have been mutated. Another example is an ASCP with a BEBD and a TEBD, and at least one CH domain selected from the group consisting of CH 1 , CH2, or CH3 in which the top region, bottom region, or any combination of top and/or bottom regions have been mutated. The SCALPs can comprise one or more constant domains (e.g., at least two, three, four, five, six, ten domains). If more than one constant domain is present in the SCALP, the domains may be of the same type or of varying types (e.g. CH 1 -CHI -CH2, CH3-CH3). The order of the single domains may be of any kind (e.g. CH1 -CH3-CH2, CH4-CH 1 -CH3 -C H2).

[0078] The BBR sequences of the Bottomside Epitope Binding Domain (BEBD)

[0079] The Bottomside Epitope Binding Domain (BEBD) of the ASCP-body comprises four regions, BBR (bottomside binding region) 1, BBR 2, and BBR 3. BBR1, BBR2 and BBR 3 are sequences which all lie within the ASCP-body sequence, but they are not adjacent to each other in the contiguous polypeptide sequence. However, because the ASCP-body folds in three-dimensional space, BBR1, BBR2, BBR3, and BBR4 together form a Bottomside Epitope Binding Domain (BEBD).

[0080] BBR1 , BBR2, BBR3, and BBR4 can be defined by their location within the consensus sequence of the ASCP-body, e.g., the consensus sequence shown in Table l ,as provided in FIG. 1A, and as provided below. FIGS 1A and B are based on pdb data bank entry 3EZJ. (Korotkov, .V., Pardon, E., Steyaert, J., Hoi, W.G. "Crystal Structure of the N-terminal domain of the secretin GspD from ETEC determined with the assistance of a nanobody". (2009) Structure 17: 255-265.) The CDR and BBR sequences are labeled and underlined below. The aligned sequences provided in Table 1 are assumed to have the corresponding CDR1, CDR2, CDR3, BBR1 , BBR2, BBR3 and BBR4 regions as designated below in the consensus sequence. Residues not underlines are framework ("FR") regions.

CONSENSUS

BBR1 CDR1 BBR2 CDR2 BBR3

QVQLV¾SGGGSVQAGGSLRLSCAASGYTYSSYCMGWFRQAP<-CTREGVfiAIWSIX3GSTYYADSVKGR

CDRl top

BBR4 CDR3

FTISQDNAKNTOTLQmSIJgEiyrA>rfYCAA^^ (SEQ ID NO: 2) top

[0081 ] BBR1 comprises the sequence at about amino acid 14 to about amino acid 17, lying between the consensus FR amino acids sequences GSVQ and LRLS. [0082] BBR2 comprises the sequence at about amino acid 40 to about amino acid 45, lying between the consensus FR amino acid sequences WFRQ and EGVA.

[0083] BBR3 comprises the sequence at about amino acid 60 to about amino acid 66, lying between the consensus FR amino acids Y and FTIS.

[0084] BBR4 comprises the sequence at about amino acid 83 to about amino acid 91, lying between the consensus FR amino acid sequences VYLQ and MYYC.

[0085] CDRI comprises the sequence at about amino acid 26 to about amino acid 33, lying between the consensus FR amino acid sequences CAAS and MGWF.

[0086] CDR2 comprises the sequence at about amino acid 49 to about amino acid 58, lying between the consensus FR amino acid sequences EGVA and YA.

[0087] CDR3 can be defined as the sequence at about amino acid 101 to about amino acid

105. CDR3 can also be defined as comprising the sequence lying between consensus FR amino acid sequences MYYCA and WGQG.

[0088] In the consensus sequence above, CDRI , CDR2, and CDR3 are GYTYSSYC,

AINSDGGSTY, and ADPGGGGGGWCLRSPLGPSGYNY (or GGGGG), respectively. BBRl, BBR2, BBR3 and BBR4 are AGGS, APGKER, ADSVKGR, and NSLKPEDTA, respectively.

[0089] Additional ASCP-body sequences, with additional CDRI , CDR2, CDR3, BBRl,

BBR2, BBR3, and BBR4 sequences are indicated in Table 1. The ASCP-bodies listed in Table 1 have previously been publically disclosed, e.g., in Verheesen et al. 2003 Biochim. Biophys. Acta 1624, 21-28; Dolk et al. 2005 Proteins 15: 555-564; De Haard et al. 2005 J. Bacteriol. 187, 4531- 4541 ; Dolk et al. Appl Environ Microbiol. 2005 71(l):442-50; Verheesen et al. 2006 Hum. Mol. Genet. 15(1), 105-1 1 1.

[0090] In another non-limiting example, a ASCP-body sequence is presented below:

QVQLQESGGGLVQAGGSLRLSCAASGSIFSIWS D DRQAPGKQRELVATITSGGSTMTYADSVKGRFTISRDNA KNTWLQI^SLCTEOTAWYCNAMVKTWAGMTRDY GQGTQVTVSS (SEQ ID NO : 3 )

[0091] The CDRI, CDR2, and CDR3 sequences are: GSIFS1NS, T1TSGGSTN, and

ANVKTWAGMTRDY. The BBRl , BBR2, BBR3, and BBR4 sequences are: AGGS; APGKQR; ADSVKGR; and NSLKPEDTA.

[0092] Methods of producing ASCP-bodies with a Bottomside Epitope Binding Domain [0093] In one embodiment, the BBR sequences of a ASCP-body of the present invention can be obtained by a method comprising the steps of: providing a V_HH domain directed to a desired antigen, either (a) by a method comprising the steps of:

(i) immunizing a mammal belonging to the Camelidae with the desired antigen or a fragment thereof, so as to raise an immune response and/or antibodies (particularly heavy chain antibodies) against the antigen; (ii) obtaining a biological sample from the immunized mammal, wherein the sample comprises heavy chain antibody sequences and/or V_HH sequences directed against the antigen; and (iii) obtaining the BBR sequences from the heavy chain antibody sequences and/or sequences; or

(b) by a method comprising the steps of

(i) screening a library comprising heavy chain antibody sequences and/or VHH sequences for sequences directed to the antigen; (ii) obtaining the heavy chain and/or V_HH sequences from the library; and (iii) obtaining the BBR sequences from the heavy chain and/or V_HH sequences.

[0094] In one embodiment, the method of obtaining the BBR sequences of a ASCP-body can further comprise the steps of (iv) subjecting the heavy chain antibody sequences and/or VHH sequences to mutagenesis (e.g., random mutagenesis or site-directed mutagenesis), to increase the affinity and/or specificity of binding to the antigen; and (v) obtaining the obtaining the mutagenized BBR sequences from the heavy chain and/or VHH sequences

[0095] In one embodiment, all BBR sequences present in a ASCP-body of the invention are derived from the same heavy chain antibody or VHH sequence. In one embodiment, it is possible to suitably combine BBRs from two or three different heavy chain antibodies or V_HH sequences against the antigen. In one embodiment, BBRs can be combined from different ASCP-bodies or different sources (e.g., synthetic BBRs or BBRs derived from a human antibody or V_H domain).

[0096] According to a non-limiting but preferred embodiment of the invention, the BEBD sequences in the ASCP-bodies of the invention are such that the ASCP-body of the invention binds to the antigen with an dissociation constant ( _D) of 10^"5 to 10^'12 moles/liter or less, and preferably 10^{" 7} to 10^'12 moles/liter or less, and more preferably 10 ^s to 10^"12 moles/liter, and/or with a binding affinity of at least 10⁷ M^'1, preferably at least 10^s M^"1, more preferably at least 10⁹ M^'1, such as at least 10^,z M^'1 and/or with an affinity less than 500 nM, preferably less than 200 nM, more preferably less than 10 nM, such as less than 500 pM. The affinity of the ASCP-body of the invention against the antigen can be determined in a manner known in the art, for example using the assay described herein.

[0097] Generation of ASCP-bodies with a BEBD using Nucleotide and Amino Acid

Substitutions [0098] ASCP-bodies of the invention having one or more amino acid or nucleotide modifications (e.g., alterations) can be generated by a variety of known methods. Typically, such ASCP-bodies are produced by recombinant methods. Moreover, because of the degeneracy of the genetic code, a variety of nucleic acid sequences can be used to encode each desired molecule.

[0099] Exemplary art recognized methods for making a nucleic acid molecule encoding an amino acid sequence variant of a starting molecule include, but are not limited to, preparation by site-directed (or oligonucleotide-mediated) mutagenesis, PCR mutagenesis, and cassette mutagenesis of an earlier prepared DNA encoding the molecule.

[00100] Site-directed mutagenesis is a preferred method for preparing substitution variants.

This technique is well known in the art (see, e.g., Carter et al. Nucleic Acids Res. 13:4431 -4443 (1985) and unkel et al, Proc. Natl. Acad. Sci. U.S.A 82:488 (1987)). Briefly, in carrying out site- directed mutagenesis of DNA, the parent DNA is altered by first hybridizing an oligonucleotide encoding the desired mutation to a single strand of such parent DNA. After hybridization, a DNA polymerase is used to synthesize an entire second strand, using the hybridized oligonucleotide as a primer, and using the single strand of the parent DNA as a template. Thus, the oligonucleotide encoding the desired mutation is incorporated in the resulting double-stranded DNA.

[00101] PCR mutagenesis is also suitable for making amino acid sequence variants of the starting molecule. See Higuchi, in PCR Protocols, pp.177-183 (Academic Press, 1990); and Valiette e/ a/., Nuc. Acids Res. 17:723-733 ( 1989). Briefly, when small amounts of template DNA are used as starting material in a PCR, primers that differ slightly in sequence from the corresponding region in a template DNA can be used to generate relatively large quantities of a specific DNA fragment that differs from the template sequence only at the positions where the primers differ from the template.

[00102] Another method for preparing variants, cassette mutagenesis, is based on the technique described by Wells et al, Gene 34:315-323 (1985). The starting material is the piasmid (or other vector) comprising the starting polypeptide DNA to be mutated. The codon(s) in the parent DNA to be mutated are identified. There must be a unique restriction endonuclease site on each side of the identified mutation site(s). If no such restriction sites exist, they may be generated using the above-described oligonucleotide-mediated mutagenesis method to introduce them at appropriate locations in the starting polypeptide DNA. The piasmid DNA is cut at these sites to linearize it. A double-stranded oligonucleotide encoding the sequence of the DNA between the restriction sites but containing the desired mutation(s) is synthesized using standard procedures, wherein the two strands of the oligonucleotide are synthesized separately and then hybridized together using standard techniques. This double-stranded oligonucleotide is referred to as the cassette. This cassette is designed to have 5' and 3' ends that are compatible with the ends of the linearized piasmid, such that it can be directly ligated to the piasmid. This piasmid now contains the mutated DNA sequence. [00103] Alternatively, or additionally, the desired amino acid sequence encoding a polypeptide variant of the molecule can be determined, and a nucleic acid sequence encoding such amino acid sequence variant can be generated synthetically. In certain embodiments, the codon usage tables for various species are incorporated to modify the nucleotide sequence for optimization of protein expression. One skilled in the art would reference the various codon optimization charts depending the species of the cells in which the ASCP-body is to be expressed.

[00104] It will be understood by one of ordinary skill in the art that the ASCP-bodies of the invention may further be modified such that they vary in amino acid sequence (e.g., from wild-type variants), but not in desired activity. For example, additional nucleotide substitutions leading to amino acid substitutions at "non-essential" amino acid residues may be made to the protein For example, a nonessential amino acid residue in a molecule may be replaced with another amino acid residue from the same side chain family. In another embodiment, a string of amino acids can be replaced with a structurally similar string that differs in order and/or composition of side chain family members, i.e., a conservative substitutions, in which an amino acid residue is replaced with an amino acid residue having a similar side chain, may be made.

[00105] Families of amino acid residues having similar side chains have been defined in the art, including basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g. , threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine).

[00106] Aside from amino acid substitutions, the present invention contemplates other modifications of the starting molecule amino acid sequence in order to generate functionally equivalent molecules. For example, one may delete one or more amino acid residues. Generally, no more than one to about ten residues will be deleted according to this embodiment of the invention. The ASCP-bodies comprising one or more amino acid deletions will preferably retain at least about 80%, and preferably at least about 90%, and most preferably at least about 95%, of the starting polypeptide molecule.

[00107] One may also make amino acid insertion variants, which retain the original ASCP- body binding molecule functionality. For example, one may introduce at least one amino acid residue (e.g. one to two amino acid residues and generally no more than ten residues) into the molecule. In another embodiment amino acid modifications may be combined within a single ASCP-body.

[00108] In one embodiment, amino acid substitutions are performed on a ASCP-body to include cysteine or other non-natural amino acid suitable for conjugating a moiety to the ASCP-body using well-known conjugating methods. In particular, the invention relates to specific amino acid variants of ASCP-bodies, wherein one or more serine amino acid residues are substituted by cysteine or a non-natural amino acid. Non-naturally occurring amino acids can be substituted into the ASCP- body using, for example, Ambrex technology (See e.g., US 7,045,337; 7,083,970).

[00109] In one embodiment, the ASCP-body is humanized using methods known in the art.

[001 10] Screening Assays for Identifying A SCP- bodies with a BEBD

[001 1 1] A variety of screening assays may be employed to identify ASCP-bodies of the invention. Essentially any in vitro or in vivo screening method that selects for binding to a desired antigen may be used.

[001 12] In one embodiment, ASCP-bodies are displayed on the surface of a cell, virus or bacteriophage and subject to selection using immobilized antigen. Suitable methods of screening are described in U.S. patent numbers 7,063,943; 6,699,658; 7,063,943 and 5866344. Such surface display may require the creation of fusion proteins of the ASCP-bodies with a suitable protein normally present on the outer surface of a cell, virus or bacteriophage. Suitable proteins from which to make such fusions are well known in the art.

[001 13] In another embodiment, ASCP-bodies are screened using an in vitro phenotype- genotype linked display such as ribosome or polysome display. Such methods of "molecular evolution" are well known in the art (see for example U.S. patent number 6, 194,550 and 7, 195,880).

[001 14] Screening methods may involve one or more in vitro or in vivo affinity maturation steps. Any affinity maturation approach can be employed that results in amino acid changes in the BBRs or the CDRs that improve the binding of the ASCP-body to the desired antigen. These amino acid changes can, for example, be achieved via random mutagenesis, "walk though mutagenesis, and "look through mutagenesis. Such mutagenesis can be achieved by using, for example, error-prone PCR, "mutator" strains of yeast or bacteria, incorporation of random or defined nucleic acid changes during ab inito synthesis of all or part of a ASCP-body. Methods for performing affinity maturation and/or mutagenesis are described, for example, in U.S. Patent Numbers 7, 195,880; 6,951 ,725;

7,078, 197; 7,022,479; 5,922,545; 5,830,721 ; 5,605,793, 5,830,650; 6,194,550; 6,699,658; 7,063,943; 5866344 and Patent Cooperation Treaty publication WO06023144. Such affinity maturation methods may further require that the stringency of the antigen-binding screening assay is increased to select for ASCP-bodies with improved affinity for antigen. Art recognized methods for increasing the stringency of a protein-protein interaction assay can be used here. In one embodiment, one or more of the assay conditions are varied (for example, the salt concentration of the assay buffer) to reduce the affinity of the ASCP-bodies for the desired antigen. In another embodiment, the length of time permitted for the ASCP-bodies to bind to the desired antigen is reduced. In another embodiment, a competitive binding step is added to the protein-protein interaction assay. For example, the ASCP-bodies are first allowed to bind to a desired immobilized antigen. A specific concentration of non- immobilized antigen is then added which serves to compete for binding with the immobilized antigen such that the ASCP-bodies with the lowest affinity for antigen are eluted from the immobilized antigen resulting in selection of ASCP-bodies with improved antigen binding affinity. The stringency of the assay conditions can be further increased by increasing the concentration of non-immobilized antigen is added to the assay.

[001 15] Screening methods of the invention may also require multiple rounds of selection to enrich for one or more ASCP-bodies with improved antigen binding. In one embodiment, at each round of selection further amino acid mutation are introduce into the ASCP-bodies. In another embodiment, at each round of selection the stringency of binding to the desired antigen is increased to select for ASCP-bodies with increased affinity for antigen.

[001 16] In the case of the bispecific ASCP-bodies of the invention, it is preferable to screen for each binding specificity independently. A first screen to identify individual ASCP-bodies that bind to a first target is performed using a first library of ASCP-bodies, where one or more amino acids in one or more of the CDRs or BBRs is altered. A second separate screen to identify individual ASCP-bodies that bind to a second target is performed using a second library of ASCP-bodies, where one or more amino acids in one or more of their CDRs or BBRs is altered. The amino acid sequences of the individual monospecific ASCP-bodies identified from both screens are determined using art recognized methods. Bispecific ASCP-bodies are generated by combining first and second target-binding sequences from individual ASCP-bodies into single, chimeric ASCP-bodies.

[001 17] Methods of Manufacture

[001 18] The ASCP-bodies of the invention are typically produced by recombinant expression. Nucleic acids encoding the molecules are inserted into expression vectors. The DNA segments encoding the molecules are operably linked to control sequences in the expression vector(s) that ensure their expression. Expression control sequences include, but are not limited to, promoters (e.g., naturally-associated or heterologous promoters), signal sequences, enhancer elements, and transcription termination sequences. Preferably, the expression control sequences are eukaryotic promoter systems in vectors capable of transforming or transfecting eukaryotic host cells. Once the vector has been incorporated into the appropriate host, the host is maintained under conditions suitable for high level expression of the nucleotide sequences, and the collection and purification of the cross-reacting ASCP-body.

[001 19] These expression vectors are typically replicable in the host organisms either as episomes or as an integral part of the host chromosomal DNA. Commonly, expression vectors contain selection markers (e.g., ampicillin-resistance, hygromycin-resistance, tetracycline resistance or neomycin resistance) to permit detection of those cells transformed with the desired DNA sequences (see, e.g., Itakura et al., U.S. Patent 4,704,362). [00120] E. coli is one prokaryotic host particularly useful for cloning the polynucleotides

(e.g., DNA sequences) of the invention. Other microbial hosts suitable for use include bacilli, such as Bacillus subtilis, and other enterobacteriaceae, such as Salmonella, Serratia, and various

Pseudomonas species.

[00121] Other microbes, such as yeast, are also useful for expression. Saccharomyces and

Pichia are exemplary yeast hosts, with suitable vectors having expression control sequences (e.g., promoters), an origin of replication, termination sequences and the like as desired. Typical promoters include 3-phosphoglycerate kinase and other glycolytic enzymes. Inducible yeast promoters include, among others, promoters from alcohol dehydrogenase, isocytochrome C, and enzymes responsible for methanol, maltose, and galactose utilization.

[00122] In addition to microorganisms, mammalian tissue culture may also be used to express and produce the polypeptides of the present invention (e.g., polynucleotides encoding immunoglobulins or fragments thereof)- See Winnacker, From Genes to Clones, VCH Publishers, N.Y., N.Y. (1987). Eukaryotic cells are actually preferred, because a number of suitable host cell lines capable of secreting heterologous proteins (e.g., intact immunoglobulins) have been developed in the art, and include CHO cell lines, various COS cell lines, HeLa cells, 293 cells, myeloma cell lines, transformed B-cells, and hybridomas. Expression vectors for these cells can include expression control sequences, such as an origin of replication, a promoter, and an enhancer (Queen et al., Immunol. Rev. 89:49 (1986)), and necessary processing information sites, such as ribosome binding sites, RNA splice sites, polyadenylation sites, and transcriptional terminator sequences. Preferred expression control sequences are promoters derived from immunoglobulin genes, SV40, adenovirus, bovine papilloma virus, cytomegalovirus and the like. See Co et al., i. Immunol.

148: 1 149 (1992).

[00123] Alternatively, coding sequences can be incorporated in transgenes for introduction into the genome of a transgenic animal and subsequent expression in the milk of the transgenic animal (see, e.g., Deboer et al., U.S. 5,741,957, Rosen, U.S. 5,304,489, and Meade et al., U.S. 5,849,992). Suitable transgenes include coding sequences for light and/or heavy chains in operable linkage with a promoter and enhancer from a mammary gland specific gene, such as casein or beta lactoglobulin.

[00124] The vectors containing the polynucleotide sequences of interest and expression control sequences can be transferred into the host cell by well-known methods, which vary depending on the type of cellular host. For example, chemically competent prokaryotic cells may be briefly heat-shocked, whereas calcium phosphate treatment, electroporation, lipofection, biolistics or viral-based transfection may be used for other cellular hosts. (See generally Sambrook et al., Molecular Cloning: A Laboratory Manual (Cold Spring Harbor Press, 2nd ed., 1989). Other methods used to transform mammalian cells include the use of polybrene, protoplast fusion, liposomes, electroporation, and microinjection (see generally, Sambrook et al., supra). For production of transgenic animals, transgenes can be microinjected into fertilized oocytes, or can be incorporated into the genome of embryonic stem cells, and the nuclei of such cells transferred into enucleated oocytes.

[00125] Once expressed, the ASCP-bodies of the present invention can be purified according to standard procedures of the art, including ammonium sulfate precipitation, affinity columns, column chromatography, HPLC purification, gel electrophoresis and the like (see generally Scopes, Protein Purification (Springer-Verlag, N.Y., (1982)). Substantially pure molecules of at least about 90 to 95% homogeneity are preferred, and 98 to 99% or more homogeneity most preferred, for pharmaceutical uses.

[00126] Methods for Grafting CDRs onto ASCP-bodies

[00127] In one aspect, the present invention features an ASCP-body altered compared to the wild-type came I id immunoglobulin to contain all or a portion of a complementarity determining region (CDR) of an antibody or a T-cell receptor.

[00128] The CDR regions of any antibody or T-cell receptor variable region, or antigen binding fragments thereof, are suitable for grafting. The CDRs can be obtained from the antibody or T-cell receptor repertoire of any animal including, but not limited to, rodents, primates, camelids or sharks. In a particular embodiment, the CDRs are obtained from CDRl , CDR2 and CDR3 of a single domain antibody, for example a ASCP-body. In a more specific embodiment, CDRl , 2 or 3 of a single domain antibody, such as a Nanobody, are grafted into an ASCP-body, thereby providing target binding specificity of the original Nanobody to the ASCP-body. Engineered libraries of camelid antibodies and antibody fragments are commercially available, for example, from Ablynx, Ghent, Belgium. The antibody repertoire can be from animals challenged with one or more antigens or from naive animals that have not been challenged with antigen. Additionally or alternatively, CDRs can be obtained from antibodies, or antigen binding fragments thereof, produced by in vitro or in vivo library screening methods, including, but not limited to, in vitro polysome or ribosome display, phage display or yeast display techniques. This includes antibodies not originally generated by in vitro or in vivo library screening methods but which have subsequently undergone mutagenesis or one or more affinity maturation steps using in vitro or in vivo screening methods. Example of such in vitro or in vivo library screening methods or affinity maturation methods are described, for example, in U.S. Patent Numbers 7,195,880; 6,951 ,725; 7,078, 197; 7,022,479; 5,922,545; 5,830,721; 5,605,793, 5,830,650; 6,194,550; 6,699,658; 7,063,943; 5866344 and Patent Cooperation Treaty publications WO06023144.

[00129] Methods to identify antibody CDRs are well known in the art (see Kabat et al., U.S.

Dept. of Health and Human Services, "Sequences of Proteins of Immunological Interest" (1983); Chothia et al., J. Mol. Biol. 196:901-917 (1987); MacCallum et al., J. Mol. Biol. 262:732-745 (1996)). The nucleic acid encoding a particular antibody can be isolated and sequenced, and the CDR sequences deduced by inspection of the encoded protein with regard to the established antibody sequence nomenclature. Methods for grafting hypervariable regions or CDRs into a ASCP-body of the invention include, for example, genetic engineering, de novo nucleic acid synthesis or PCR-based gene assembly (see for example U.S. patent number 5,225.539).

[00130] The above techniques allow for the identification of a suitable selection and presentation of a CDR. However, additional metrics can be invoked to further improve the fit and presentation of the hypervariable region based on structural modeling of the ASCP-body and the donor antibody.

[00131 ] In one aspect, specific amino acid residues in any of the beta-strands or alpha-helix of an ASCP-body are mutated to allow the CDR loops to adopt a conformation that retains or improves binding to antigen. This procedure can be performed in a way that is analogous to grafting a CDR into a heterologous antibody framework, i.e., using a combination of structural modeling and sequence comparison. In one embodiment, the ASCP-body residues adjacent to a CDR are mutated in a similar manner to that performed by Queen et al. (see U.S. patent numbers 6,180,370; 5,693,762; 5,693,761; 5,585,089; 7,022,500). In another embodiment, ASCP-body residues within one Van der Waals radius of CDR residues are mutated in a similar manner to that performed by Winter et al. (see U.S. patent numbers 6,548,640; 6,982,321). In another embodiment, ASCP-body residues that are non-adjacent to CDR residues but are predicted, based upon structural modeling of the ASCP-body and the donor antibody, to modify the conformation of CDR residues are mutated in a similar manner to that performed by Carter et al. or Adair et al (see U.S. patent numbers 6,407,213;

6,639,055; 5,859,205; 6,632,927).

[00132] Libraries of ASCP-bodies comprising an epitope binding domain comprising BBR1,

BBR2, BBR3, andBBR4.

[00133] In another aspect, the invention provides a library of ASCP-bodies comprising an epitope binding domain comprising BBR1, BBR2, BBR3, and BBR4 , which can be used to identify ASCP-bodies which bind to a particular desired target. In one embodiment, the library comprises ASCP-bodies, each of which contains at least one amino acid alteration in BBR1, BBR2, BBR3, and/or BBR4.

[00134] Library diversity can be generated by, for example, random mutagenesis, "walkthrough mutagenesis", or "look-through mutagenesis" of any of the ASCP-body sequences disclosed herein. Nucleic acids encoding the library of ASCP-bodies, or variants thereof, described herein can be constructed using art-recognized methods including, but not limited to, PCR-based or enzyme- mediate genetic engineering, ab initio DNA or RNA synthesis, and/or cassette mutagenesis. Alternatively, ASCP-bodies can be generated by combining BBR1, BBR2 and/or BB 3 sequences from one or more different library members.

[00135] A library of bi-specific ASCP-bodies can also be produced. An ASCP-body has two potential epitope binding domains. The bottomside epitope binding domain (BEBD) comprises four bottomside binding regions: BBR1, BBR2, BB 3, and BBR4. The topside epitope binding domain (TEBD) comprises three complementarity determining regions: CDR1, CDR2, and CDR3.

[00136] A library of bi-specific ASCP-bodies can be generated by combining BEBD sequence(s) from one or more ASCP-bodies with TEBD sequence(s) from one or more ASCP- bodies. Alternatively, bi-specific ASCP-bodies can also be created by first identifying a monospecific ASCP-body with a desired function, e,g., a TEBD that binds to human serum albumin. This molecule is then used as a scaffold; the loops of the opposite face (e.g., the BEBD) of the ASCP- body are varied to generate the library. This library of bi-specific ASCP-bodies is then used to screen against the second target molecule to identify a bi-specific entity that binds to two targets. In one non-limiting example, a mono-specific ASCP-body can bind to HSA using the BEBD. This mono-specific ASCP-body is used as a scaffold to generate a library in which at least one or more of the CDRs is varied such that it binds to a second target, e.g., VEGFR2. This library of bi-specific ASCP-bodies can be used to screen for the VEGFR2 target.

[00137] Additionally or alternatively, CDRs are obtained from antibodies, or antigen binding fragments thereof, produced by in vitro or in vivo library screening methods, including, but not limited to, in vitro polysome or ribosome display, phage display or yeast display techniques. This includes antibodies not originally generated by in vitro or in vivo library screening methods but which have subsequently undergone mutagenesis or one or more affinity maturation steps using in vitro or in vivo screening methods. Examples of such in vitro or in vivo library screening methods or affinity maturation methods are described, for example, in U.S. Patent Numbers 7, 195,880;

6,951 ,725; 7,078, 197; 7,022,479; 5,922,545; 5,830,721 ; 5,605,793, 5,830,650; 6,194,550; 6,699,658; 7,063,943; 5866344 and Patent Cooperation Treaty publications WO06023144.

[00138] Library Construction

[00139] A BEBD sequence disclosed herein (such as one of those disclosed in Table 1 ) can be used as the scaffold to generate libraries of ASCP-bodies. See also Fig. IB.

[00140] Using computational modeling, the BBR1, BBR2 and/or BBR3 of the BEBD sequence are chosen to be randomized. The DNA sequences corresponding to the variegated BBR sequences are optimized (based on codon frequency) for expression in E.coli. Regions are synthesized as degenerated positions. The libraries are assembled from synthetic degenerated oligonucleotides and genes corresponding to full-length fragments gel-purified. Amplification can be performed with terminal primers and subsequent ligation of the amplified library into cloning vector pCR-Script to yield the ASCP-body libraries. The libraries are then screened to identify ASCP- bodies with desired binding characteristics.

[00141] A method of screening for ASCP-bodies generated from the above-described libraries is described herein. The libraries are subcloned into a yeast display vector such as pYDl (Invitrogen) using homologous recombination methods and transformed into a suitable strain such as EBY100 using standard molecular biology techniques.

[00142] If a ASCP-body specific to human serum albumin is desired, presentation and selection of ASCP-bodies against human serum albumin is conducted, following essentially the protocol previously published by Lipovsek, D. et al, (J. Mol. Biol. 2007 May 1 1 ; 368(4): 1024-41 ). The libraries are screened for binders to human serum albumin.

[00143] ASCP-bodies with a preferred binding affinity (e.g., binding to lysozyme, or human serum albumin) can be identified from a library using magnetic bead sorting. Yeast cultures presenting a library of ASCP-bodies are induced, for example, for 18 h at 30°C in galactose- containing medium (90% SG-CAA 10% SD-CAA, 50 μ&ΊηΙ. kanamycin, 100 U/mL penicillin G, 200 U/mL streptomycin). Induced yeast library cells (e.g., 10⁹ cells) are washed, e.g., with 25 mL of ice-cold phosphate-buffered saline (PBS), pH 7.4, 2 mM ethylenediaminetetraacetic acid (EDTA), and 0.5% bovine serum albumin (BSA), and then incubated, e.g., in 5 mL of the same buffer containing 1 μΜ biotinylated hen egg white lysozyme (HEL-b, Sigma, St. Louis, MO) for 1 h at room temperature with gentle rotation. Following the incubation, the sample is chilled, washed and resuspended, e.g., chilled on ice, washed, e.g., with 25 mL of ice-cold PBS, pH 7.4, 2 mM EDTA, and 0.5% BSA and resuspended, e.g., in 2.5 mL of the same buffer. Magnetic beads [e.g., 100-μί aliquot of magnetic Streptavidin MicroBeads (Miltenyi Biotec, Auburn, CA)] are added to the yeast and incubated, e.g., on ice for 10 min. A solution [e.g., ice-cold PBS, pH 7.4, 2 mM EDTA, 0.5% BSA] is added to the sample, e.g., to a total volume of 25 mL, immediately before the cells are subjected to separation, e.g., on an AutoMACS Cell Separator (Miltenyi Biotec), using the preset program for positive selection of rare cells (possel s). Selected cells are collected, e.g., in 6 mL SD- CAA, pH 4.5, 50 μξ/πύ_^ kanamycin, 100 U/mL penicillin G, 200 U/mL streptomycin; quantified, e.g., by serial dilution followed by plating on SD-CAA agar plates; and grown, e.g., in 50 mL of the same medium for 2 days at 30°C, to identify cells expressing a desired ASCP-body.

[00144] In one non-limiting embodiment, ASCP-bodies with a preferred binding affinity

(e.g., binding to lysozyme or human serum albumin) can be identified from a library using fluorescence-activated cell sorting. Subsequent rounds of selection are performed by FACS, e.g., starting with 2 ^χ 10⁶ to 3 x 10⁶ induced yeast cells. Cells are washed, e.g., with 1 mL PBS, pH 7.4, and 0.1% BSA, resuspended, e.g., in 100 uL of the same buffer containing biotinylated hen egg white lysozyme, and incubated, e.g., at room temperature with gentle rotation for 1 h. After being washed, e.g., with 1 mL of ice-cold PBS, pH 7.4, 0.1% BSA, the cells were labeled, e.g., with antibodies and streptavidin. Mouse monoclonal FITC-conjugated anti-c-myc antibody (AbD Serotec) can be used to label the yeast for surface display of c-myc-tagged antibody mimics, and PE- labeled streptavidin (Invitrogen) or anti-biotin antibody (Miltenyi) can be used to label HEL-b associated with lysozyme-binding antibody mimics. The FACS sorts are performed on yeast cells labeled with FITC-conjugated mouse anti-c-myc antibody and PE-conjugated streptavidin

(Invitrogen).

[00145] Double- labeled yeast cells are sorted, e.g., on a Dako MoFlo high-speed cell sorter with a 488 nm laser, at 6000-10,000 cells/s. Gates are adjusted to collect the desired yeast cells, e.g., cells with the highest 0.1-1% of HEL-b-associated signal (PE) and in the top half of expression- associated signal (F1TC). Duplicate samples are labeled with the same antibody and streptavidin reagents, but in the absence of HEL-b were used to avoid selecting the cells that bound detection reagents instead of lysozyme.

[00146] For all libraries, the first two FACS sorts are performed on labeled yeast cells, e.g., labeled with 1 μΜ HEL-b. Once a population of cells is observed that was labeled with PE in the presence but not in the absence of HEL-b, the concentration of HEL-b in the subsequent round can be decreased by an order of magnitude. Selected cells are collected, e.g., in 0.5 mL of SD-CAA, pH 4.5, 50 μ Ληί kanamycin, 100 TJ/mL penicillin G, and 200 U/mL streptomycin. The collected cells were grown, e.g., to saturation in 5 mL of the same medium, with shaking, for 2 days at 30 °C, before being induced and labeled for the next round of sorting.

[00147] After several rounds of FACS sorting the final enriched population is plated out, e.g., on SDCAA plates and incubated at 30°C for 2 days. Individual colonies are picked, e.g., using a Genetix Clonepix, and re-arrayed, e.g., into 96 well plates containing SD-CAA medium. After incubation, e.g., for 24 hours, the cells are collected, e.g., by centrifugation, and re-suspended, e.g., in SD-GAA medium, for induction of surface expressed ASCP-bodies. Positive clones can be identified by standard ELISA. Plasmid DNA corresponding to the unique ASCP-bodies can be purified and sequenced to identify mono-specific binders.

[00148] Once mono-specific binders are identified and selected from a library, they can be used independently to generate therapeutic molecules. In particular, the mono-specific binders generated from a library using the BEBD of a ASCP-body can be used to generate novel therapeutic binding molecules against a target of interest. Various mono-specific binders from a library can be combined with linkers to produce a ASCP-body that is capable of binding to one or more regions of a single target (e.g., TNF). Alternately, a ASCP-body comprising binders from a library can also be designed to bind to one or more regions of multiple targets (e.g., one or more regions of HSA and TNF).

[00149] In addition, the mono-specific binders generated from a library can be combined using standard molecular biology technique to generate bi-specific and multi-specific. [00150] Bi-specific and multi-specific ASCP-bodies comprising an epitope binding domain comprising BBR1, BBR2, BBR3, and BBR4.

[00151] The present invention also encompasses a ASCP-body or conjugate of two or more

ASCP-bodies which bind to two distinct antigen targets of interest. In one embodiment of a single bi-specific ASCP-body, the TEBD binds to one antigen, while the BEBD binds to a different antigen. In another embodiment, the "bi-specific" composition comprises a fusion of two or more ASCP-bodies. In this embodiment, at least one binding domain (e.g., either a BEBD or TEBD) binds to one antigen, while another binding domain (e.g., either a BEBD or a TEBD, preferably on a different ASCP-body) binds to a second antigen target. For example, a bi-specific composition can be a ASCP-body with a BEBD that binds to one target, fused to a second ASCP-body with a TEBD that binds to another target. In another embodiment, the bi-specific composition can be a ASCP- body with a BEBD that binds to one target, fused to a second ASCP-body with a BEBD that binds to another target.

[00152] In one embodiment, the present invention encompasses a "multi-specific" composition that binds to more than one target. For example, various mono-specific ASCP-bodies (each of which bind to a single, different target) can be conjugated, forming a composition capable of binding to more than two targets simultaneously. A multi-specific composition also includes, for example, one ASCP-body with a BEBD that binds one target and a TEBD that binds to a second target, fused to a second ASCP-body with a BEBD that binds to a third target. The TEBD of the second ASCP-body can optionally target a fourth target.

[00153] ASCP-bodies of the present invention can be linked in various ways. A pair of

ASCP-bodies may be linked to each other. Several ASCP-bodies may be linked like pearls on a chain.

[00154] Multispecific ASCP-bodies can be produced using art recognized methods. For example, ASCP-bodies may be linked genetically, such that multispecific ASCP-bodies are expressed as a single polypeptide. This linkage may be direct or conferred by an additional amino acid "linker" sequence. Suitable non-limiting methods and linkers are described, for example, in US20060286603 and WO04041862A2. Exemplary polypeptide linkers include, but are not limited to, GS linkers, such as GGGGSGGGGS (SEQ ID NO:4), GSGSGSGSGS (SEQ ID NO:5), PSTSTST (SEQ ID NO:6), and EIDKPSQ (SEQ ID NO: 7), and multimers thereof.

[00155] The multispecific ASCP-bodies generated using linker sequences have an improved steric hindrance for binding to target molecules, thus enabling shorter linker sequences to be used to link two or more monomeric ASCP-bodies together. Shorter linker sequences cause less immunogenic responses and are less likely to get cleaved. [00156] Alternatively, multispecific ASCP-bodies may be prepared by chemically conjugating the individual ASCP-bodies using methods known in the art. A variety of coupling or cross-linking agents may be used for covalent conjugation. Examples of cross-linking agents include, e.g., protein A, carbodiimide, N-succinimidyi-S-acetyl-thioacetate (SATA), 5,5'-dithiobis(2- nitrobenzoic acid) (DTNB), o-phenylenedimaleimide (oPDM), N-succinimidyl-3-(2- pyridyldithio)propionate (SPDP), and sulfosuccinimidyl 4-(N-maleimidomethyl) cyclohaxane-1 - carboxylate (sulfo-SMCC) (see e.g., Karpovsky et al. ( 1984) J. Exp. Med. 160: 1686; Liu, MA et al. (1985) Proc. Natl. Acad. Sci. U.S.A 82:8648). Other methods include those described in Paulus (1985) Behring Ins. Mitt. No. 78, 1 18-132; Brennan et al. ( 1985) Science 229:81-83), and Glennie et al. (1987) J. Immunol. 139: 2367-2375). Preferred conjugating agents are SATA and sulfo-SMCC, both available from Pierce Chemical Co. (Rockford, 1L). Cysteine residues can be introduced into the ASCP-bodies at specific positions and then crosslink with reagents to sulfhydryl such as DPDPB or DTME (available from Pierce) to link two molecules together. Additional methods pertaining to HSA fusions can be found, for example, in WO 2001077137 and WO 200306007. Alternatively, the constituent molecules can be encoded in the same vector and expressed as a single protein in a host cell. Methods for producing such fusion proteins are described, for example, in U.S. Patent Number 5,260,203; U.S. Patent Number 5,455,030; U.S. Patent Number 4,881, 175; U.S. Patent Number 5,132,405; U.S. Patent Number 5,091,513; U.S. Patent Number 5,476,786; U.S. Patent Number 5,013,653; U.S. Patent Number 5,258,498; and U.S. Patent Number 5,482,858.

[001 7] Similar methods known in the art may be used to conjugate a ASCP-body to a non-

ASCP-body entity.

[00158] In yet another embodiment, the invention provides ASCP-bodies that are conjugated to polyethylene glycol (PEG), for example, to increase the biological (e.g., serum) half-life of the molecule. Methods for PEGyiating proteins are well known in the art. For example, the ASCP-body can be reacted with a PEG moiety, such as a reactive ester or aldehyde derivative of PEG, under conditions in which one or more PEG groups become attached to the molecule. The term

"PEGylation moiety", "polyethylene glycol moiety", or "PEG moiety" includes a polyalkylene glycol compound or a derivative thereof, with or without coupling agents or derivatization with coupling or activating moieties (e.g., with thiol, triflate, tresylate, azirdine, oxirane, or preferably with a maleimide moiety, e.g., PEG-maleimide). Other appropriate polyalkylene glycol compounds include, but are not limited to, maleimido monomethoxy PEG, activated PEG polypropylene glycol, but also charged or neutral polymers of the following types: dextran, colominic acids, or other carbohydrate based polymers, polymers of amino acids, and biotin derivatives.

[00159] The choice of the suitable functional group for a PEG derivative is based on the type of available reactive group on the molecule that will be coupled to the PEG. For proteins, typical reactive amino acids include lysine, cysteine, histidine, arginine, aspartic acid, glutamic acid, serine, threonine, tyrosine. The N-terminal amino group and the C-terminal carboxylic acid can also be used.

[00160] Preferably, the pegylation is carried out via an acylation reaction or an alkylation reaction with a reactive PEG molecule (or an analogous reactive water-soluble polymer). As used herein, the term "polyethylene glycol" is intended to encompass any of the forms of PEG that have been used to derivatize other proteins, such as mono (Ci-Cio) alkoxy- or aryloxy-polyethylene glycol or polyethylene glycol-maleimide. Methods for pegylating proteins are known in the art and can be applied to the present invention. See for example, WO 2005056764, U.S.7,045,337, U.S.7,083,970, U.S.6,927,042, EP 0 154 316 by Nishimura et al. and EP 0 401 384 by Ishikawa et al. ASCP-bodies can be engineered to include at least one cysteine amino acid or at least one non-natural amino acid to facilitate pegylation.

[001 1 ] Binding of the ASCP-body conjugates to their specific targets can be confirmed by various assays, for example, the fusion can be radioactively labeled and used in a radioimmunoassay (RIA) (see, for example, Weintraub, B., Principles of Radioimmunoassays, Seventh Training Course on Radioligand Assay Techniques, The Endocrine Society, March, 1 86, which is incorporated by reference herein). The radioactive isotope can be detected by such means as the use of a gamma- counter or a scintillation counter or by autoradiography.

[00162] Other ASCP-body conjugates of the present invention include a ASCP-body linked to a tag (e.g., biotin) or a chemical (e.g., an immunotoxin or chemotherapeutic agent). Such chemicals include a cytotoxic agent, which is any agent that is detrimental to (e.g., kills) cells.

Examples include taxol, cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicin, doxorubicin, daunorubicin, dihydroxy anthracin dione, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, and puromycin and analogs or homologs thereof.

Therapeutic agents also include, e.g., antimetabolites (e.g., methotrexate, 6-mercaptopurine, 6- thioguanine, cytarabine, 5-fluorouracil decarbazine), alkylating agents (e.g., mechlorethamine, thioepa chlorambucil, melphalan, carmustine (BSNU) and lomustine (CCNU), cyclothosphamide, busulfan, dibromomannitol, streptozotocin, mitomycin C, and cis-dichlorodiamine platinum (II) (DDP) cisplatin), anthracyclines (e.g., daunorubicin (formerly daunomycin) and doxorubicin), antibiotics (e.g., dactinomycin (formerly actinomycin), bleomycin, mithramycin, and anthramycin (AMC)), and anti-mitotic agents (e.g., vincristine and vinblastine). Other examples of therapeutic cytotoxins that can be conjugated to ASCP-body of the invention include duocarmycins, calicheamicins, maytansines and auristatins, and derivatives thereof.

[00163] Cytotoxins can be conjugated to the ASCP-bodies of the invention using linker technology available in the art. Examples of linker types that have been used to conjugate a cytotoxin include, but are not limited to, e.g., hydrazones, thioethers, esters, disulfides and peptide- containing linkers. A linker can be chosen that is, for example, susceptible to cleavage by low pH within the lysosomal compartment or susceptible to cleavage by proteases, such as proteases preferentially expressed in tumor tissue such as cathepsins (e.g., cathepsins B, C, D). For further discussion of types of cytotoxins, linkers and methods for conjugating therapeutic agents, see also Saito, G. et al. (2003) Adv. Drug Deliv. Rev. 55: 199-215; Trail, P.A. et al. (2003) Cancer Immunol. Immunother. 52:328-337; Payne, G. (2003) Cancer Cell 3:207-212; Allen, T.M. (2002) Nat. Rev. Cancer 2:750-763; Pastan, I. and Kreitman, R. J. (2002) Curr. Opin. Investig. Drugs 3: 1089-1091 ; Senter, P.D. and Springer, C.J. (2001) Adv. Drug Deliv. Rev. 53:247-264.

[00164] The ASCP-bodies of the present invention also can be linked to a radioactive isotope to generate cytotoxic radiopharmaceuticals, also referred to as radioimmunoconjugates. Examples of radioactive isotopes that can be conjugated to ASCP-bodies for use diagnostically or therapeutically include, but are not limited to, iodine¹³¹, indium¹¹¹, yttrium⁹⁰ and lutetium¹⁷⁷. Methods for preparing radioimmunoconjugates are established in the art. Examples of radioimmunoconjugates are commercially available, including ibritumomab, tiuxetan, and tositumomab, and similar methods can be used to prepare radioimmunoconjugates using the molecules of the invention.

[00165] The ASCP-body conjugates of the invention can be used to modify a given biological response, and the drug moiety is not to be construed as limited to classical chemical therapeutic agents. For example, the drug moiety may be a protein or polypeptide possessing a desired biological activity. Such proteins may include, for example, an enzymatically active toxin, or active fragment thereof, such as abrin, ricin A, pseudomonas exotoxin, or diphtheria toxin; a protein such as tumor necrosis factor or interferon-y; or, biological response modifiers such as, for example, lymphokines, interleukin-1 ("IL-1 "), interleukin-2 ("IL-2"), interleukin-6 ("IL-6"), granulocyte macrophage colony stimulating factor ("GM-CSF"), granulocyte colony stimulating factor ("G-CSF"), or other growth factors.

[00166] Techniques for conjugating such therapeutic moiety are well known and can be applied to the molecules of the present invention, see, e.g., Arnon et al., "Monoclonal Antibodies For Immunotargeting Of Drugs In Cancer Therapy", in Monoclonal Antibodies And Cancer Therapy, Reisfeld et al. (eds.), pp. 243-56 (Alan R. Liss, Inc. 1985); Hellstrom et al., "Antibodies For Drug Delivery", in Controlled Drug Delivery (2nd Ed.), Robinson et al. (eds.), pp. 623-53 (Marcel Dekker, Inc. 1987); Thorpe, "Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A Review", in Monoclonal Antibodies '84: Biological And Clinical Applications, Pinchera et al. (eds.), pp. 475-506 (1985); "Analysis, Results, And Future Prospective Of The Therapeutic Use Of Radiolabeled Antibody In Cancer Therapy", in Monoclonal Antibodies For Cancer Detection And Therapy, Baldwin et al. (eds.), pp. 303-16 (Academic Press 1985), and Thorpe et al., "The Preparation And Cytotoxic Properties Of Antibody-Toxin Conjugates", Immunol. Rev., 62: 1 19-58 (1982). [00167] The present AS CP-bodies also can be modified by hesylation, which utilizes hydroxyethyl starch ("HES") derivatives linked to drug substances in order to modify the drug characteristics. HES is a modified natural polymer derived from waxy maize starch that is metabolized by the body's enzymes. This modification enables the prolongation of the circulation half-life by increasing the stability of the molecule, as well as by reducing renal clearance, resulting in an increased biological activity. Furthermore, HESylation potentially alters the immunogenicity or allergenicity. By varying different parameters, such as the molecular weight of HES, a wide range of HES drug conjugates can be customized.

[00168] Patent publications DE 196 28 705 and DE 101 29 369 describe possible methods for carrying out the coupling of hydroxyethyl starch in anhydrous dimethyl sulfoxide (DMSO) via the corresponding aldonolactone of hydroxyethyl starch with free amino groups of hemoglobin and amphotericin B, respectively. Since it is often not possible to use anhydrous, aprotic solvents specifically in the case of proteins, either for solubility reasons or else on the grounds of denaturation of the proteins, coupling methods with HES in an aqueous medium are also available. For example, coupling of hydroxyethyl starch which has been selectively oxidized at the reducing end of the chain to the aldonic acid is possible through the mediation of water-soluble carbodiimide EDC ( l-ethyl-3- (3-dimethyl-aminopropyl)carbodiimide) (PCT EP 02/02928). Additional hesylation methods which can be applied to the present invention are described, for example, in patent publications U.S.

20070134197, U.S. 20060258607, U.S. 20060217293, U.S. 20060100176, and U.S.20060052342.

[00169] The present ASCP-bodies also can be modified via sugar residues. Methods for modifying sugar residues of proteins or glycosylating proteins are known in the art (see, for example, Borman (2006) Chem. & Eng. News 84(36): 13-22 and Borman (2007) Chem. & Eng. News 85: 19- 20) and can be applied to the molecules of the present invention.

[00170] Additionally or alternatively, ASCP-bodies can be made that have an altered type of glycosylation, such as a hypofucosylated pattern having reduced amounts of fucosyl residues or an AS CP-body having increased bisecting GlcNac structures. Such carbohydrate modifications can be accomplished by, for example, expressing the ASCP-body in a host cell with altered glycosylation machinery. Cells with altered glycosylation machinery have been described in the art and can be used as host cells in which to express recombinant ASCP-bodies of the invention to thereby produce ASCP-bodies of the invention with altered glycosylation. For example, EP 1 , 176, 195 by Hang et al. describes a cell line with a functionally disrupted FUT8 gene, which encodes a fucosyl transferase, such that antibodies expressed in such a cell line exhibit hypofucosylation. PCT Publication WO 03/035835 by Presta describes a variant CHO cell line, Lecl3 cells, with reduced ability to attach fucose to Asn(297)-linked carbohydrates, also resulting in hypofucosylation of antibodies expressed in that host cell (see also Shields, R.L. et al., 2002 J. Biol. Chem. 277:26733-26740). Methods to produce polypeptides with human-like glycosylation patterns have also been described by

EP1297172B 1 and other patent families originating from Glycofi.

[00171] Compositions

[00172] The ASCP-body binding of the present invention have in vivo therapeutic utilities.

Accordingly, the present invention also provides compositions, e.g., a pharmaceutical composition, containing one or a combination of ASCP-bodies (or variants, fusions, and conjugates thereof), formulated together with a pharmaceutically acceptable carrier. Pharmaceutical compositions of the invention also can be administered in combination therapy, i.e., combined with other agents. For example, the combination therapy can include a composition of the present invention with at least one or more additional therapeutic agents, such as anti-inflammatory agents, anti-cancer agents, and chemotherapeutic agents.

[00173] The pharmaceutical compositions of the invention can also be administered in conjunction with radiation therapy. Co-administration with other ASCP-bodies are also

encompassed by the invention.

[00174] As used herein, "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible. Preferably, the carrier is suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal or epidermal administration (e.g., by injection or infusion). Depending on the route of administration, the active compound, i.e., antibody, bispecific and multispecific molecule, may be coated in a material to protect the compound from the action of acids and other natural conditions that may inactivate the compound.

[00175] A "pharmaceutically acceptable salt" refers to a salt that retains the desired biological activity of the parent compound and does not impart any undesired toxicological effects (see e.g., Berge, S.M., et al. (1977) J. Pharm. Sci. 66: 1-19). Examples of such salts include acid addition salts and base addition salts. Acid addition salts include those derived from nontoxic inorganic acids, such as hydrochloric, nitric, phosphoric, sulfuric, hydro bromic, hydroiodic, phosphorous and the like, as well as from nontoxic organic acids such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxy alkanoic acids, aromatic acids, aliphatic and aromatic sulfonic acids and the like. Base addition salts include those derived from alkaline earth metals, such as sodium, potassium, magnesium, calcium and the like, as well as from nontoxic organic amines, such as Ν,Ν'-dibenzylethylenediamine, N-methylglucamine,

chloroprocaine, choline, diethanolamine, ethylenediamine, procaine and the like. [00176] An ASCP-body can be administered by a variety of methods known in the art. As will be appreciated by the skilled artisan, the route and/or mode of administration will vary depending upon the desired results. The active compounds can be prepared with carriers that will protect the compound against rapid release, such as a controlled release formulation, including implants, transdermal patches, and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Many methods for the preparation of such formulations are patented or generally known to those skilled in the art. See, e.g., Sustained and Controlled Release Drug Delivery Systems, J.R. Robinson, ed., Marcel Dekker, Inc., New York, 1978.

[00177] To administer an ASCP-body by certain routes of administration, it may be necessary to coat the compound with, or co-administer the compound with, a material to prevent its inactivation. For example, the compound may be administered to a subject in an appropriate carrier, for example, liposomes, or a diluent. Pharmaceutically acceptable diluents include saline and aqueous buffer solutions. Liposomes include water-in-oil-in-water CGF emulsions as well as conventional liposomes (Strejan et al. (1984) J. Neuroimmunol. 7:27).

[00178] Pharmaceutically acceptable carriers include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. The use of such media and agents for pharmaceutically active substances is known in the art. Except insofar as any conventional media or agent is incompatible with the active compound, use thereof in the pharmaceutical compositions of the invention is contemplated. Supplementary active compounds can also be incorporated into the compositions.

[00179] Therapeutic compositions typically must be sterile and stable under the conditions of manufacture and storage. The composition can be formulated as a solution, microemulsion, liposome, or other ordered structure suitable to high drug concentration. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride in the composition. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent that delays absorption, for example, monostearate salts and gelatin.

[00180] Sterile injectable solutions can be prepared by incorporating the active compound in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by sterilization microti ltration. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze-drying (lyophilization) that yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.

[00181] Dosage regimens are adjusted to provide the optimum desired response (e.g., a therapeutic response). For example, a single bolus may be administered, several divided doses may be administered over time or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. For example, the ASCP-body may be administered once or twice weekly by subcutaneous injection or once or twice monthly by subcutaneous injection.

[00182] It is especially advantageous to formulate parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the subjects to be treated; each unit contains a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specification for the dosage unit forms of the invention are dictated by and directly dependent on (a) the unique characteristics of the active compound and the particular therapeutic effect to be achieved, and (b) the limitations inherent in the art of compounding such an active compound for the treatment of sensitivity in individuals.

[00183] Examples of pharmaceuticatly-acceptable antioxidants include: (1 ) water soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol, and the like; and (3) metal chelating agents, such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

[00184] For the therapeutic compositions, formulations of the present invention include those suitable for oral, nasal, topical (including buccal and sublingual), rectal, vaginal and/or parenteral administration. The formulations may conveniently be presented in unit dosage form and may be prepared by any methods known in the art of pharmacy. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will vary depending upon the subject being treated, and the particular mode of administration. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will generally be that amount of the composition which produces a therapeutic effect. Generally, out of one hundred per cent, this amount will range from about 0.001 per cent to about ninety percent of active ingredient, preferably from about 0.005 per cent to about 70 per cent, most preferably from about 0.01 per cent to about 30 per cent. [O0185] Formulations of the present invention that are suitable for vaginal administration also include pessaries, tampons, creams, gels, pastes, foams or spray form lations containing such carriers as are known to be appropriate. Dosage forms for the topical or transdermal administration of ASCP-body compositions include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. The active compound may be mixed under sterile conditions with a pharmaceutically acceptable carrier, and with any preservatives, buffers, or propellants which may be required.

[00186] The phrases "parenteral administration" and "administered parenterally" as used herein means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural and intrasternal injection and infusion.

[00187] Examples of suitable aqueous and nonaqueous carriers which may be employed in the pharmaceutical compositions of the invention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating materials, such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.

[00188] These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of presence of microorganisms may be ensured both by sterilization procedures, supra, and by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents, such as sugars, sodium chloride, and the like into the compositions. In addition, prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents which delay absorption such as aluminum monostearate and gelatin.

[00189] When the compounds of the present invention are administered as pharmaceuticals, to humans and animals, they can be given alone or as a pharmaceutical composition containing, for example, O.O01 to 90% (more preferably, O.0O5 to 70%, such as 0.01 to 30%) of active ingredient in combination with a pharmaceutically acceptable carrier.

[0O190] Regardless of the route of administration selected, the ASCP-body of the present invention, which may be used in a suitable hydrated form, and/or the pharmaceutical compositions of the present invention, are formulated into pharmaceutically acceptable dosage forms by conventional methods known to those of skill in the art. [00191] Actual dosage levels of the active ingredients in the pharmaceutical compositions of the present invention may be varied so as to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, mode of administration, and composition, without being toxic to the patient. The selected dosage level will depend upon a variety of pharmacokinetic factors including the activity of the particular compositions of the present invention employed, or the ester, salt or amide thereof, the route of administration, the time of administration, the rate of excretion of the particular compound being employed, the duration of the treatment, other drugs, compounds and/or materials used in combination with the particular compositions employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts. A physician or veterinarian having ordinary skill in the art can readily determine and prescribe the effective amount of the pharmaceutical composition required. For example, the physician or veterinarian could start doses of the compounds of the invention employed in the pharmaceutical composition at levels lower than that required in order to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. In general, a suitable daily dose of a compositions of the invention will be that amount of the compound that is the lowest dose effective to produce a therapeutic effect. Such an effective dose will generally depend upon the factors described above. It is preferred that administration be intravenous, intramuscular, intraperitoneal, or subcutaneous, preferably administered proximal to the site of the target. If desired, the effective daily dose of therapeutic compositions may be administered as two, three, four, five, six or more sub-doses administered separately at appropriate intervals throughout the day, optionally, in unit dosage forms. While it is possible for a compound of the present invention to be administered alone, it is preferable to administer the compound as a pharmaceutical formulation (composition).

[00192] Therapeutic compositions can be administered with medical devices known in the art. For example, in a preferred embodiment, a therapeutic composition of the invention can be administered with a needleless hypodermic injection device, such as the devices disclosed in U.S. Patent Nos. 5,399,163, 5,383,851, 5,312,335, 5,064,413, 4,941 ,880, 4,790,824, or 4,596,556.

Examples of well-known implants and modules useful in the present invention include: U.S. Patent No. 4,487,603, which discloses an implantable micro-infusion pump for dispensing medication at a controlled rate; U.S. Patent No. 4.,486, 194, which discloses a therapeutic device for administering medicants through the skin; U.S. Patent No. 4,447,233, which discloses a medication infusion pump for delivering medication at a precise infusion rate; U.S. Patent No. 4,447,224, which discloses a variable flow implantable infusion apparatus for continuous drug delivery; U.S. Patent

No. 4,439, 196, which discloses an osmotic drug delivery system having multi-chamber

compartments; and U.S. Patent No. 4,475, 196, which discloses an osmotic drug delivery system. Many other such implants, delivery systems, and modules are known to those skilled in the art. [00193] In certain embodiments, the molecules of the invention can be formulated to ensure proper distribution in vivo. For example, the blood-brain barrier (BBB) excludes many highly hydrophilic compounds. To ensure that the therapeutic compounds of the invention cross the BBB (if desired), they can be formulated, for example, in liposomes. For methods of manufacturing liposomes, see, e.g., U.S. Patents 4,522,81 1 ; 5,374,548; and 5,399,331. The liposomes may comprise one or more moieties which are selectively transported into specific cells or organs, thus enhance targeted drug delivery (see, e.g., V.V. Ranade (1989) J. Clin. Pharmacol. 29:685).

Exemplary targeting moieties include folate or biotin (see, e.g., U.S. Patent 5,416,016 to Low et al.); mannosides (Umezawa et al., (1988) Biochem. Biophys. Res. Commun. 153: 1038); antibodies (P.G. Bloeman et al. (1995) FEBS Lett. 357: 140; M. Owais et al. (1995) Antimicrob. Agents Chemother. 39: 180); surfactant protein A receptor (Briscoe et al. (1995) Am. J. Physiol. 1233: 134), different species of which may comprise the formulations of the inventions, as well as components of the invented molecules; pl20 (Schreier et al. (1 94) J. Biol. Chem. 269:9090); see also K. Keinanen (1994) FEBS Lett. 346: 123; JJ. Killion (1994) Immunomethods 4:273. In one embodiment, the therapeutic compounds of the invention are formulated in liposomes; in a more preferred embodiment, the liposomes include a targeting moiety. In one embodiment, the therapeutic compounds in the liposomes are delivered by bolus injection to a site proximal to the tumor or infection. The composition must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi.

[00194] In one embodiment, the molecules of the invention can be formulated to prevent or reduce the transport across the placenta. This can be done by methods known in the art, e.g., by PEGylation of the ASCP-body. Further references can be made to "Cunningham-Rundles C, Zhuo Z, Griffith B, Keenan J. ( 1992) J Immunol Methods. 152: 177-190; and to "Landor M. (1995) Ann Allergy Asthma Immunol 74:279-283. This is particularly relevant when the ASCP-body are used for treating or preventing recurrent spontaneous abortion.

[001 5] The ability of a compound to inhibit cancer can be evaluated in an animal model system predictive of efficacy in human tumors. Alternatively, this property of a composition can be evaluated by examining the ability of the compound to inhibit, such inhibition in vitro by assays known to the skilled practitioner. A therapeutically effective amount of a therapeutic compound can decrease tumor size, or otherwise ameliorate symptoms in a subject. One of ordinary skill in the art would be able to determine such amounts based on such factors as the subject's size, the severity of the subject's symptoms, and the particular composition or route of administration selected.

[00196] The composition must be sterile and fluid to the extent that the composition is deliverable by syringe. In addition to water, the carrier can be an isotonic buffered saline solution, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. Proper fluidity can be maintained, for example, by use of coating such as lecithin, by maintenance of required particle size in the case of dispersion and by use of surfactants. In many cases, it is preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol or sorbitol, and sodium chloride in the composition. Long-term absorption of the injectable compositions can be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate or gelatin.

[00197] When the active compound is suitably protected, as described above, the compound may be orally administered, for example, with an inert diluent or an assimilable edible carrier.

[00198] Therapeutic and Diagnostic Applications

[00199] The ASCP-bodies described herein may be constructed to bind any antigen or target of interest. Such targets include, but are not limited to, cluster domains, cell receptors, cell receptor ligands, growth factors, interleukins, protein allergens, bacteria, or viruses. The ASCP-bodies described herein may also be modified to have increased stability and half-life, as well as additional functional moieties. Accordingly, these molecules may be employed in place of antibodies in all areas in which antibodies are used, including in the research, therapeutic, and diagnostic fields. In addition, because these molecules possess solubility and stability properties superior to antibodies, the ASCP-bodies herein may be used under conditions that would destroy or inactivate antibody molecules.

[00200] For example, ASCP-bodies can be administered to cells in culture, e.g. in vitro or ex vivo, or in a subject, e.g., in vivo, to treat, prevent or diagnose a variety of disorders. The term "subject" as used herein in intended to includes human and non-human animals. Non-human animals includes all vertebrates, e.g., mammals and non-mammals, such as non-human primates, sheep, dogs, cats, cows, horses, chickens, amphibians, and reptiles. When the ASCP-bodies are administered together with another agent, the two can be administered in either order or

simultaneously.

[00201] In one embodiment, the ASCP-bodies (and variants, fusions, and conjugates thereof) of the invention can be used to detect levels of the target bound by the molecule and/or the targets bound by a bispecific/multispecific ASCP-body. This can be achieved, for example, by contacting a sample (such as an in vitro sample) and a control sample with the molecule under conditions that allow for the formation of a complex between the molecule and the target(s). Any complexes formed between the molecule and the target(s) are detected and compared in the sample and the control. For example, standard detection methods, well-known in the art, such as ELISA, FACS, and flow cytometric assays, can be performed using the compositions of the invention.

[00202] Also within the scope of the invention are kits comprising the compositions (e.g.,

ASCP-bodies, variants, fusions, and conjugates thereof) of the invention and instructions for use. The kit can further contain a least one additional reagent, or one or more additional ASCP-bodies of the invention (e.g., an ASCP-body having a complementary activity that binds to an epitope on the target antigen distinct from the first molecule). Kits typically include a label indicating the intended use of the contents of the kit. The term label includes any writing, or recorded material supplied on or with the kit, or which otherwise accompanies the kit.

[00203] As described above, the molecules of the present invention may be employed in all areas of the research, therapeutic, and diagnostic fields. Exemplary diseases/disorders which can be treated using the ASCP-bodies of the present invention (and variants, fusions, and conjugates thereof) include autoimmune disorders, cancers, infections, and other pathogenic indications.

[00204] Specific examples of autoimmune conditions in which the ASCP-bodies of the invention can be used include, but are not limited to, the following: multiple sclerosis and other demyelinating diseases; rheumatoid arthritis; inflammatory bowel disease; systemic lupus erythematosus; Type I diabetes; inflammatory skin disorders; Sjogren's Syndrome; and transplant rejection.

[00205] Specific examples of cancers in which the ASCP-bodies can be used include, but are not limited to, the following: lung; breast; prostate; bladder; melanoma; non-Hodgkin lymphoma; colon and rectal; pancreatic; endometrial; kidney; skin (non-melanoma); leukemia; and thyroid.

[00206] The ASCP-bodies can be used for the treatment of prevention of hyperproliferative diseases or cancer and the metastatic spread of cancers. Non- limiting examples of cancers include bladder, blood, bone, brain, breast, cartilage, colon kidney, liver, lung, lymph node, nervous tissue, ovary, pancreatic, prostate, skeletal muscle, skin, spinal cord, spleen, stomach, testes, thymus, thyroid, trachea, urogenital tract, ureter, urethra, uterus, or vaginal cancer. As described herein, angiogenesis-associated diseases include, but are not limited to, angiogenesis-dependent cancer, including, for example, solid tumors, blood born tumors such as leukemias, and tumor metastases; benign tumors, for example hemangiomas, acoustic neuromas, neurofibromas, trachomas, and pyogenic granulomas; inflammatory disorders such as immune and non-immune inflammation; chronic articular rheumatism and psoriasis; ocular angiogenic diseases, for example, diabetic retinopathy, retinopathy of prematurity, macular degeneration, corneal graft rejection, neovascular glaucoma, retrolental fibroplasia, rubeosis; Osier- Webber Syndrome; myocardial angiogenesis; plaque neovascularization; telangiectasia; hemophiliac joints; angiofibroma; and wound granulation and wound healing; telangiectasia psoriasis scleroderma, pyogenic granuloma, cororany collaterals, ischemic limb angiogenesis, corneal diseases, rubeosis, arthritis, diabetic neovascularization, fractures, vasculogenesis, hematopoiesis (see e.g., WO2005056764).

[00207] Specific examples of infections in which the ASCP-bodies of the invention can be used include, but are not limited to, the following: cellular, fungal, bacterial, and viral. [00208] Unless indicated otherwise, all methods, steps, techniques and manipulations that are not specifically described in detail can be performed and have been performed in a manner known per se, as will be clear to the skilled person. Reference is for example again made to the standard handbooks and the general background art mentioned herein and to the further references cited therein.

[00209] Claims to the invention are non-limiting and are provided below.

At_00091 (SEQ ID N0:#)

DVQLVESGGGSVQAGK3SLRLSCAASGYTASTYCMAWFRQAPGKEREGVATINRSS . STYYDVSVKGRFWSQDNAKNTVYLQMNSLKPEDTAIYYCAAIEEYCGGTYW....LRPNKYKHWGQGTQVTVSS Ca_00046 (SEQ ID N0:#)

AVQLVESGGGSVQTGGSLRLSCVASGYTYSSARIG FRQAPGKEREGVAAILTDGV TYYADAVKGRFTISRDNAKNTVYLQMNSLKPEDTAMYYCA FRVSQYNGWGQGTQVTVSS

Ca_00177 (SEQ ID N0:#)

QVQLVESGGGSVQAGGSLRLSCAASGYIYSTNYMAWFRQAPGKEREGVARIYFGG3TPYYADSVKGRFTM_^ LSSSGYQYWGQGTQVVSS

Ll_„00095 (SEQ ID N0:#)

QVQLQESGGGLVQPGGSLRLSCAASGGTFS YAMGWFRQAPGKEREFVATVSRGGGSTYYADSVKGRFTISRDNAKNTVYLQMNSLKPEDTAAYYCAAGRGSPSDTG RPDEYDYWGQGTQVTVSS

Ca_00317 (SEQ ID N0:#)

QVQLVESGGGSVQAGGSLRLSCAASGYTYSNFCMAWFRQAPGQAREGVAYIYTGGNS . YYADSVKG FTISQDNAKNTVYLMNSLKPEDTAIYYCAADNGPGTCIGR PEYLFRYWGRGTQVTVSS

Ca_00026 (SEQ ID N0:#>

QVQLVESGGGSVQTGGSLRLSCAASGYTYTRRCMA FRQAPGKEREGVALIYIDGGRTDYADSAKGRFTISQDRANTVYLQtOTSLKPEDTA ^ ....KAYAYRYWGQGTQVTVSS

At_00089 (SEQ ID N0:#)

QVQLVESGGGSVQTGGSLRLSCAASGYTYTRRCMAWFRQAPGKEREGVALIYIDGGRTDYADSAKGRFTISQDRAKNTWLQ^ KAYAYRYWGQGTQVTVSS

Ca_00047 (SEQ ID N0:#)

QVQLVESGGGSVQAGGSLRLSCAASGYTISGICMGWFRQAPGKERECTAGISPGGTGTYYADSVK^ .... RGNGYRYWGQGTQVTVSS

Ll_00342 (SEQ ID N0:#)

QVQLQESGGGLVQAGGSLTLSCAASGRTFSSHSMGWFRQAPGKEREFVATIIRSDGSTYYADSVKGRFTISREMAKWTVYLQiraSLKPEDTAVYYCAARTW RGHEYNYWGQGTQVTVSS

Ca_00307 (SEQ ID N0:#)

QVKLEESGGGSVQAGGSLRLSCATSGYTYS-MCIGWFRQAPGKEREGVAVIYTggSSTFYADSV GRFTISQDNANTVYLQM SLKPEDTAMYYCAAVGYTDYCKS PWPEAYRFWGQGTQVTVSS

Ca_00175 (SEQ ID N0:#)

DVQLVESGGGSVHAGGSLRLSGAVSGYTGStyCIAVffRQAPGKEREGVATINSSGGSRYSADSVKGRFTLSQDNAKKTVYLH™ . , , EPAYNYWGQGTQVTVSS

Ll_00705 (SEQ ID N0:#[

DVQLQASGGGLVQAGGSLRLSCAASGRTFSSDAMGWFRQAPGKEREFVAAISWSGGSTYYADSVKGRFTISR YPPSEYDYWGQGTQVTVSS

Ca_00103 (SEQ ID N0:#)

DVQLVESGGGSVQAGGSLRLSCAASGVTYSS YMGWFRQAPGKEREGVAVICTDGRSTYYADSVKGRFAVSLDNA NTVSLQINSLK™

Ll_00751 (SEQ ID N0:#)

QVQLQESGGGLVQAGGSLRLSCAASGFTSDDYAVGWFRQAPGKEREGVSCISSGIX5STYYADSVKGRFTISTDNASNT^

LI_00003 (SEQ ID N0:#)

QVQLQESGGGLVQAGGSLP^SCAASGFTSDDYAVGWFRQAPGKEREGVSCISSGIXSSTYYADSVKGPJTISTDNASOT

Ca_00019 (SEQ ID N0:#)

QVQLVESGGGSVQAGGSLRLSCAASGYTFSSYPMGWYRQAPGKECELVSRIFSIX;. SA YAGSVKGRFTISRIOTAKKTAYLQ1IDSLKPEDTAVYY ..TCYGPNYWGQGTQVTVSS

At_00086 (SEQ ID N0:#)

QVQLVESGGGSVQAGGSLRLSCAASGYTFSSYPMGWYRQAPGKECELVSRIFSDG. SANYAGSVKGRFTISRDNA NTAYLQMDSLKPEDTAVYYCAAGPGSG LWAG ...TCYGPNYWGQGTQVTVSS Ca_00165 (SEQ ID N0:#)

QVKLEESGGGSVQAGGSLRLSCAASGDTYSSHCMGWFRQAPGKEREGVAAI .. GGGLTYYADSVKGRFTISQDNRKNTVYLQMTSLKPEDTAMYYCAARGGGCWYPW WDAYHYWGQGTQVTVSS

Ca_00090 (SEQ ID N0:#)

QVQLVESGGGSVQAGGSLRLSCAASGYTYRSICMGWFRQAPGKEREGVAVINSDRDYPYYADSVKGRFTISQDNAK^^ ....KELKYNYWGQGTQVTVSS

Ll„01134 (SEQ ID NO:#)

QVQLQESGGGLVQAGGSLRLSCIASGRTFSSYAMGWFRQAPGKEREFLATINWSGGTTYYADSVKGRFTVSRIMAK^^ DSKSFDYWGQGTQVTVSS

Ca_00146 (SEQ ID N0:#)

EVQLVESGG43SVQAGGSLRLSCAASGYSFSGYCMGWFRQVPGKEREGVATIOTFGGTTLYADSVKARFTISRDNA ...GESGYFYWGRGTQVTVSS

Ll_00099 (SEQ ID N0:#)

QVQLQQSG<¾LVQA∞SLTLSCAASGRTFSSHSMGWFRQAPGKEREFVATIIRSDGSTYYADSVKGPJTISRDN^ ....RGHEYNYWGQGTQVTVSS

Ll_00343 (SEQ ID N0:#)

QVQLQQSGGGLVQAGGSLTLSCAASGRTFSSHSMGWFRQAPGKEREFVATIIRSDGSTYYADSVKGRFTISRDNAKNW ... RGHEYNYWGQGTQVTVSS

Ca_00360 (SEQ ID N0:#)

AVQLVESGGGSVQAGGSLKLSCAVSGYTySSGCMGWFRQAPGKEREGVATIDSGGG AFYADSVKGRFIISQDHVQNTVYLQMNNLKPEDTAIYYCAADPKLVCGSVAPTILLFRLLsSYWGQGTQVTVSS Ca_00064 (SEQ ID N0:#)

EVQLVESGG3SVQAGGSLRLSCAGSGFTYSTNCMGRFRQVPGKEREGVATIYTRRCSTYYADSVKGRFTISQDNAK^ LRGVDFGWWGQGTQVTVSS

44 Ca_00327 (SEQ ID N0:#)

Q QLVESGGGSVQAGGSLRLSCOTSGFSGSTACMGWFRQAPGKEREGLAIINRRGGSAYYADSVKGRFTISQDVGKN^ GVDDFHEWGQGTQVTVSS

Ca_00039 (SEQ ID N0:#)

AVQLVDSGGGSVQAGGSLRLSCAASGTTYCTYDIAWYRQAPEKDYEFVSVIDSDG. STRYADSV GRFTISRDNAKNTVYLQMNSLKPEDTAMYYCKTVFKSWCSDG LGTTLPNYWGQGTQVTVSS

Ca_QQ284 (SEQ ID N0:#)

QVQLVESGGGSVQAGGSLRLSCAASAYTDsyYCIGWRQAPGOTjPEGVAYITTRAGTTIYADSV GRFTISQDKAKNTSYLQOTSLTP

Ll„00741 (SEQ ID N0:#)

EVQLQASGGGLVQAGDSLRLSCAASGRTFSSYHMGWFRQAPGKEMFVAAINWSGDTTYYE^SVKGR^^ ...AEGDYGYWGQGTQVTVSS

Ll_01133 (SEQ ID N0:#)

QVQLQDSGGGLVQAGGSLRLSCAAsgYTFDDYAIGWFRQAPGKEREGVSCISSSDGTTYYADSVKGRFTISSDNA^ HPRYDYWGQGTQVTVSS

Ll_01132 (SEQ ID N0:#)

QVQLQDSGGGLVQAGESLRLSCAASERTFSTYAMGWFRQAPGKEREFVAGISGSGGRTYYEDSVKGRFTISRDNAKSMV^^ ... YRASDFDSWGQGTQVTVSS

Ca„00040 (SEQ ID N0:#)

DVQLVESGGGSVQAGGSLRLSCAASGATYCTYDMSWYRQAPEKDYEFVSVIDSDG. SARYADSVKGRFTISRDNAKNTVYLQMNSLKPEDTGMYYCK VFKSWCSDG LGTTLPNYWGQGTQVTVSS

Ca_00185 (SEQ ID N0:#)

EVQLVESGGGSVQAGGSLRLSCVVSGVTNSRYSIGWFRQAPEKEREAVAAINTGGM . TNYVTSVKGRFTISQDNAKNTVYLQMNSL PDDTAIYYCALGYRSYYRDYA .... LLSDGYNSWGQGTQVSVSS Ll„00712 (SEQ ID N0:#)

DVQLQASGGGLVQAGGSLRLSCAASGLTFSSYVMGWRQAPGKERDFVAAIITSGRSTYYADSVKGRFTISRDNAKNTVY^ RPADYNYWGQGTQVTVSS

Ca_00099 (SEQ ID N0:#)

DCQLVESGGGSVQAGGSLRLSCAASG ..YDTYC GWRQAPGKERDWAVINTGGGSIAYHDSVKGRFTISQDimNTWLQKNSLKPEDTAIYYCVAARPHAYCNGVT APLNFLSWGQGTQVTVSS

Ca_00194 (SEQ ID N0:#)

QVQLVESGGGSVQAGGSLRLSCVASAN ..WYSVGWFRLAPGKEREOTATI RGSFITYYADSV GRFTISHDNA NTWLQ^SL SEDTAIYYCASKPVYKSGHWT DDRGYTYWGQGTQVTVSΞ

Ll„00672 (SEQ ID N0:#)

QVQLQDSGGGLVQAGGSLRLSCAASGFTFDDWIGWFRQAPGKEREGVSCIWSSEGSTYYADS GRFWSSDNAKHTWLQIOTSLKPEDTAVYYCAAAG-^SYHYTC ... DPLYG DYWGKGTLVTVSS Ca_00206 (SEQ ID N0:#)

QVQLTOSGGGSVQAGGSLRLSCAVSGYTGRPYSMAWFRQAPGKEREGVATIYKIGPISYY HirreGRFTISQDNAEM DVSAYTYWGQGTQVTVSS

Ll_00406 (SEQ ID N0:#)

QVQLQESGGGLVQAGGSLRLSCAAARGTFSTYAMGWFRQAPGKAREFVAGISRSGSMTHYADSVKGRFTISRDNAKN^ ... SSSRDYDYWGQGTQVTVSS

Ca_00364 (SEQ ID N0:#)

QVQLVESGGSSVQAGGSLRLSCTASGDTDNLYSLS^RQAPGKEREGVAGIRRGGGNPKYSDSVKGRFTISQOT^ ... PNQYNYWGQGTQVTVSS

Ll_01131 (SEQ ID N0:#)

QVQLQESGGGLVQAGGSLRLSCAASGRTFSSYAMGWFRQAPGKEREFVAGISGSGGRTYYEDSVKGRFTITRDNGKSV^ ... RASDFDSWGQGTQVTVSS

L1_Q0005 (SEQ ID N0:#)

QVQLQESGGGLVQAGGSLPJ-SCAASGFTFDDAAIGWRQAPGKEREAVSCISTSDSSTYYADSVKGRFTISSDNAKNTM^^

Ll_QQ115 (SEQ ID N0:#)

QVQLQESGGGLVQAGGFLPJSCAASGFTFNHYlOTGWFRQAPGTERELVAVIDRPGroSTYYVESVKGRFTISSDNAF-KTVDLQMNSLK .. ELRPSDYDY GQGTQVTVSS

Ll„00708 (SEQ ID N0:#)

EVQLQASGGGLVQAGGSLRLSCTASGRTFSTYTMGWFRQAPGKEREFVAAISPJSSVGTYYRD-WKGRFTISRD^ STRYNYWGQGTQVTVSS

Ll_00649 (SEQ ID N0:#)

QVQLQDSGGGLEQAGGSLRLSCAVSGRTSSTYAMGWFRQAPGKEREFVAAISWSGGSIHYADSVKGRFTISRDNAKSTVYLQMNSLKPEDTAVYYCAGNFAAWVGRNNAYI .. RGDEYMYWGQGAQVTVSS Ca„00081 (SEQ ID N0:#)

EVQLVESGGGSVQAGGSLRLSCAASGISNRRFCMAWFQPAPGKE EGVAFIERGGSTTLYADSVKGRFTISTDNAKNTMALQMNSLKPEDTAIYYCAANGRIWGLGSWWN ... KHDAYNFWGQGTQVTVSS Ca_00222 ( SEQ ID NO: #)

AVQLVESGGGSVQAGGSLRLSCAASGFTYSDSSIAVffRQVPGKEREGVARI TTSGDTYHAASVKGRF ISQ ...FKEKNYNAWGQGTQVTVSS

Ll_00023 (SEQ ID NO:#)

QVQLQESGGGLVQPGGSLRLSCATSGFTFGDYDMSWVRQAPRQGPEWVSTI RDGSSAYYEDSVKGRF ISRDNA NTVYLQMNSLKPEDTAVYSCAAGRDYGMGYTTGP ..LRSSDFGSWGQGTQVTVSS Ll_00318 (SEQ ID NO:#)

QVQLQESGGRLVEAGGSLRLSCAVSGGTFSWYAMGWFRQAPGKEREFVATVSRGGGASYYADSVKDRFTISRDNAKNTV^ RPDEYDYWGQGTQVTVSS

At„00016 (SEQ ID NO:#)

QVQLQESGGGLVQAGGSLRLSCAASGRATSgyGMGWFRQVPGKEREFVAAIRWSGWTYYVDSVKGRFTISRDNAKSTVYLQm LPVGFDYWGQGTQVTVSS

 Ca_00184 (SEQ ID N0:#)

QVQLVESGGDSVQAGGSLRLSCAASGYTIRPYCMGWFRQAPGKEREGVAVMTIGGTIPFYADSVKGRFTISQDTAKNTVYLQMDSLLPKDTAIYYCAARSGFCRRE FEHDAYDNWGQGTQVTVSS

At_Q0Q02 (SEQ ID N0:#)

QVQLQESGGGLVQAGGSLRLSCVASGRTFsgYGMGWFRQVPGKEREFVAAIRWSGKETWYVDSVKGRFTISRD SPVGFDYWGQGTQV VSS

Ll_01013 (SEQ ID N0:#)

... LQXSGGGLVQAGGSLRLSC¾VSGGTFSWYAMGWFRQAPGKEREFVATISRGGGSSYYADSVKGRFTISF RPDEYDYWGQGTQV ...

L1_Q0Q53 (SEQ ID N0:#)

QVQLQQSGGGLVQAGGSLRLSCAASGRISS .NAMAWFRQNPGKERDFVATISWNGGSTDYADSLKGRF ISRDNAKNTVYLQMNSLKPEDTAVYYCAAALGKPRGAKE DARGYDYWGQGTQVTVSS

Ca_00052 (SEQ ID N0:#)

EVQLVESGGGSVQAGGSLRLSCAAPGNTYSTNLMGWFRQAPGKEREGVAAI( GRGTTF GTSQY YWGQGTQVTVSS

Ca_Q035S (SEQ ID N0:#)

DVQLVESGGGSVQAGGSLRLSCTVSGNTYDAYCVAVffRQGPG EREGVARAHSNGGGTYIADSVKGRFTLSQDHAKSTAYLQMNSLK ... PSDGYQYWGQGTQVTVSS

Ll_00381 (SEQ ID N0:#)

QVQLQQSGGGLVQAGGSLRLSCAASGRTFSSYPMGWFRQAPG EREFVAAIGRLVTFTRYTDSVKGRF ISRDNAKDTMYLQMNNLKPEDTAVYYCAAFRGHNLFSRY ....AQVTDYDYWGQGTQVTVSS Ll_Q0375 (SEQ ID N0:#)

QVQLQQSGGGLVQAGGSLRLSCAASGRTFSSYPMGWFRQAPGKEREFVAAIGRLV FTRYTDSVKGRFTISRDNAKDTMYLQMNNLKPEDTAVYYCAAFRGHNLFSRY AQVTDYDYWGQGTQVTVSS

Ca_00203 (SEQ ID NO : # )

AVQLVDSGGDSVQAGGSLRLSCVASGYTGKSYCLG FRQPPGKEREWVAAKDSGGPNTYYADSVKGRFTIPQGST .LVYLQMNSLKPEDTAIYYCAAARLPEEGTC . SNLRFSSDDFEYWGQGTQVIVSS Ll_00385 (SEQ ID NO:#)

, ... SETEYDYWGQGTQVTVSS SPVGFDYWGQGTQVTVSS

....NSDAYNYWGQGTQVTVSS FSRSDYDYWGQGTRVTVSS DPNNYHYWGQGTQV VSS

... DPARYAYWGQGTQVTISS LPVGFDYWGQGTQVTVSS

...LDAAEFHLWGQGTQVTVSS Ll_00669 (SEQ ID NO:#)

QVQLQESGGGLVQAGGSLRLSCAASGRPFSDYVMGV^RQAPGKERDFVAAI -WSGGVTYYADSVKGRFTIS N RAYDTWGQGTQVTVSS

Ca_00145 (SEQ ID NO:#)

AVQLVESGGGSVQAGGSLRLSCTASGVGFSFYTMAWFHQAPGKECELVSIISLDG. TTNY DSVKGRFTISRDNGKNTADLQMNSLILEDTGVYYCAAGVVSGTVVLDTC. PPRSHESAYWGQGTQVTVSS Ll_OQ658 (SEQ ID NO:#)

QVQLQDSGGGLVQAGGSLRLSCAASGRPFSNYAVGWFRQAPGKEREFVAAISRILGWTYYTDSVGRFTISRDNA STVYLQrOTSLNPEDTAVYYCAARLDFN YS SDYDYWGQGTQVTVSS

Ca_OQ 13 (SEQ ID NO:#!

AVQLVESGGGSVPAGGSL LSCAASGYTFGIYPMAWVRQAPGKECELVATLGSDG. STNYYDSAKGRFTISRDNAKNTVYLQKNSLKPEDTARYYCAADGPDDIATPCPC.GAQYRFLEVWGQGTLVNVSS Ll_00268 (SEQ ID NO:#)

QVQLQQSGGGLVQAGGFLRLSCAASGRAFNLYAMGWFRQAPGKEREFVATIIWTGGTSSYADSVKGRFNISRDNAKN^ VRSRAYDYWGQGTQVTVSS

Ll_00744 (SEQ ID NO:#)

...LQESGGGLVQAGGSLRLSCAASGFTSDDYAIGWFRQAPGKEREGVSCISSSDGSTYYADSVKGRFTISSDKAKHTVYLQMN^

L1_0Q0Q9 (SEQ ID NO:#|

QVQLQESGGGLVQPGESLRLSCAASGFTFDDYAIVWFRQAPGKEREGVSCISTNGDYTYYADSVKDRFTISRDNAKIiTVYLQMSSLKPEDTAVYYCAAVYDY ... GTPDNMNYWGKGTLVTVSS

At_00011 (SEQ ID NO:#)

QVQLQQSGGGLVQAGGSLRLSCVASGRTF sgYGMGWFRQVPEKEREFVAAIRWSGVTTYYVDSVKGRFTISRDNAKSTVYLQMNSLKPEDTAVYYCAVRPVRVD IS SPVGFDYWGQGTQVTVSS

 At_00009 (SEQ ID N0:#)

QVQLQQSGGGLVQAGGSLRLS AASGRATSgyGMG FRQVPGKEREFVAAIR SGWTYYVDSVKGRFTISRDKAKTTVYLQ LPVGFDYWGQGTQVTVSS

Ll_00368 (SEQ ID N0:#)

QVQLQESGGGLVQAGGSLRLSMSGRl^RRnE-MGWFRRAPGKEREFVAAISWSGGATSYKDSVKGRFTISRDNA STLYLQKDS .... PHSEYEYDYWGQGTQVTVSS

Ll_01006 (SEQ ID N0:#)

... LQESGGRLVEAGGSLRLSCAVSGGTFSWYAMGWRQAPGKEREFVATISRGGGSSYYADSVKGRFTISR RPDEYDYWGQGTQV ...

At_^OOOOS (SEQ ID NO:#)

QVQLQQSGGGLVQAGGSLRLSCVASGRTFsgYGMGWFRQVPGKEREFVAAIRWSGKETWYVDSVKGRFTISRIWAKTT^ SPVGFDY GQGTQV VSS

Ca_00348 (SEQ ID NO:#)

AVQLVDSGGGSVQAGGSLRLSCttSQYMFDTRAMAWFRQPPGKECELVSIIKSDG . STGVADSWGRFAISQDSNENTVYLC^MSLKPEDTAWYCAADVGLRLTPDDVLT GGPrdFRY GQGTQV VSS Ca_00303 (SEQ ID NO:#)

QVQLVESGGGSVQAGGSLRLSCTASGFIFDASDMGWYHQVPGNECELVSTISSKG.NTYYADSVKGRFTISRDNAKNTM^^ LGTTRSDCWGQGTQVTVYS

At_00015 (SEQ ID NO:#)

QVQLQESGGGLVQAGGSLRLSCAASGRASsgYGMGWFRQVPGKEREFVAAIRWSGKETWYKDSWGRFTISR^ SPVGFDYWGQGTQVTVSS

Ll_00534 (SEQ ID NO:#)

...LQASGGGLVQAGDSLRLSCtaSGLTFSTYAMGWFRQAPGKEREFVAAINRGG ISHYADSVKGRFT^ ..

Ll_00871 (SEQ ID NO:#)

... LQASGGGLVQAGDSLRLSCtaSGLTFSTYAMGWFRQAFGKEREFVAAINRGGKISHYADSVKGF-F^ ...

Ca_00176 (SEQ ID NO:#)

DVQLVESGGSSVQAGGSLRLSCAASGYASSSPFMGWFRQAFGKEREAVAVTYIiDGATKYCVNAVKGRFTI^ HRSLY YWGQGTQVTVSS

Ll_00044 (SEQ ID NO:#)

QVQLQQSGG3LVQTGGSLRLSCAASGRtyGGYC¾GV^RQVPGKERELVAAIRWSGISTYYADSVKGRFTISRDN^ SPVGFAYWGQGTQVTVSS

Ll_00757 (SEQ ID NO:#)

QVQLQQSGGGLVQTGGSLRLSCAASGRtyGGYGMGWFRQVPGKERELVAAIRWSGISTYYADSVKGRFTISRDN^ SPVGFAYWGQGTQVTVSS

Ca_00110 (SEQ ID NO:#)

QVQLVESGGGSVQAGGSLRLSCAVSGAgySSRFMAV^RQ PGKEREGIACISPNGLDTFYRDSV^ .WASTSPYDSWGQGTQVTVSS Ca_0O296 (SEQ ID NO:#)

QVQLVESGGGSVQAGGSLTLSCVASGNIYCDYDMNWYRQAPGKEREFVSAIDSDG . STNYADSVKGRFTISQDKALDPVYLQ NGLKPEDTAMYICKFECYSTDYGGNV... TAGPGLGYWGQGTQVIVSS Ca_00263 (SEQ ID NO:#)

QVQLVESGGGSVQAGGSLRLSC SGYsgSNNCMCH^RQAFGKEREGVATIAGRIXSSTWYGNSV^ ... EAHGYNYWGRGTQVNVSS

Ll_010 9 (SEQ ID NO:#)

... LQESGGGLVQAGGSLRLSCAASGRDFSTYALAWFRQAFGKETOFVAAITWTGGSTYYADS PPRYDYWGQGTQVT...

At_00025 (SEQ ID NO:#)

QVQLQESGGGNVTAGGSLRLSCAASGRtyGGYGMGWFRQVPGKERELVAAIRWSGTSTYY DSVKGRFTISRDNVKNMVYLQMNSLKPEDTAVYHCAARTVRWDIS SPVGFAYWGQGTQVTVSS

Ll_00668 (SEQ ID NO:#)

QVQLQESGGGLVQAGGSLRLSCAASGRPFSDYVMGWFRQAPGKERDFVAAITWSGGVTYYADSLKGRFTISNDNAKKTVLLQMNSL PEDTAV^ NKRAYDT GQGTQVTVSS

Ca_00272 (SEQ ID NO:#)

AVQLVESGGGSVQAGGSLRLSCTVSGYTY DYCAGWFRQVPGEARFJtVAFINSGGGKTKYADSVKGR^^ .... GRYDYHAWGQGSQVTVSS

Ca_00379 (SEQ ID NO:#)

QVQLVESGGGSVQAGGSLRLSCTVSGWISSRYCLIWFRQAPGKEQEGVASINTGGGVTYYSDSVKGRFTISQGNTKNTWYLQMNNLKPEDTSIYTCGT , FLVGFDDLVCRPDVSIYPSKFWGQGLQVTVSS Ca_00259 (SEQ ID NO:#)

QVQLVESGGGSVQAGGSLRLSCKHIGYLFSRHAMGWFRHRPGKECELVSMVLSDG . TTDYADSVKGRFTISRDNAKHTVDLQMDSLKPEDTAVYYCAADNPGPPVCRGGNCKVCDTQAGYFGQGTQVTVSS Ll_00930 (SEQ ID NO:#)

QVQLQESGGGLVRTGGSLRLSOU^GRTLSSTTVAWFRQAPGKDRELVTAVKWTGISTYYADSVKGR^ PSQYDYRGQGTQVTVSS

Ll_00932 (SEQ ID NO:#)

QVQLQESGGGLVRTGGSLRLSCAASGRTLSSTTVAWFRQAPGKDRELVTAVKWTGISTYYADSVKGRFTISR^ PSQYDYRGQGTQVTVSS

Ll_00931 ( SEQ ID NO : # )

QVQLQESGGGLVRTGGSLRLSCAASGRTLSSTTVAWFRQAPGKDRELVTAVKWTGISTYYADSVKGRFTI^ PSQYDYRGQGTQVTVSS

Ll_0093 (SEQ ID NO:#)

QVQLQESGGGLVRTGGSLRLSCAASGRTLSSTWAWFRQAPG DRELVTAVKWTGISTYYADSVKGRFTISRIWV NTWLQMNSLKPEDTAVYY PSQYDYRGQGTQVTVSS

Ll_00933 (SEQ ID N0:#)

QVQLQESGGGLVRTGGSLRLSCMSGRTLSSTWAV^IQAPGKDRELVTAVKOTGISTYYADSVKGRF PSQYDYRGQGTQVTVSS

Ll_00544 (SEQ ID N0:#)

QVQLQDSGGGLVQAGK5SLRLSCWPTRTISTYAMGWFRQAPGKEREFVAAINWRTGRTYYEDSVEGR^^ ...DPARYAYWGQGTQVTVSS

Ca_QQ269 (SEQ ID N0:#)

QVQLVESGGGSVQAGGSLRLSCVITEYSYSTYCMG FRQAPGKDREGVASINSATGTTYYADSVKGRF ITQ.. IKNTVYLQMTSLTPEDTAIYYCAADSASSVGFCSN . , .. PLEDMDYWGKGTLVTISS Ca_00127 (SEQ ID N0:#)

QVQLVESGGGSVQAGGSLKLSCAASGYTGTTCRMAWYRQAPGKERVLVSSITSHGD . TYYAESV GRFTISQDNDKKTVYFQK SLKPEDTAKYLCNTIACPAGVSCTRC.. YSEPEFWHWGQGTQVTVSS Ca„00347 (SEQ ID N0:#)

AVQLVESGGGSVQAGGSLTLSCVASGYHFSTDCMAWFRQAPGNepeeVASIDKggGLPDyaDS™ GPRGYNYWGQGTQVTVSS

Ca_00178 (SEQ ID N0:#)

EVQLVESGGGSVQAGGSLRLSCTTSNYALGDVCTGWFRQAPGKEREGVAAIytGGGA^ APASDFTYWGQGTQVTVSS

Ll„00055 (SEQ ID N0:#)

QVQLQESGGGLVQAGGSLALSCAASGLLFSSYDMGWFRQFPGKEREFVAAISASGRR DYVDSVKGRF VSRDVAKPΓVFLQMDLKPED AVYYCAGGKSAYRGGA....HYTTGAYDYWGQGTQVTVSS Ca_00190 (SEQ ID N0:#)

QVQLVESGGGSVQAGGSLELSCRASGYIΥΞ ... I^WFRQAPGKEREGVA YRGTOITYYADSVKGRFSISQVTAENSVYLQlfflSLKPEDTAIYYCAARLSYNSLHWT NLWAYKAWGQGTQVTVSS

Ll_00707 (SEQ ID NO:#)

EVQLQASGGGSVQTGASLRISCLASGLPFSTYSKGWYRQAPGKEREFVAVI .GGGGNTYHADSLKDRFTISRDNDKNTVYLQKNSL PEDTAVYYCAADRDF IVAGFI SQYSPRAVEYWGEGTQVTVSS Ca_00164 (SEQ ID N0:#)

AVQLVESGGGSVQAGGSLRLSCAASGYDHCTYDMSWYRQAPGKEREFVSTIDSDDMIT. YAESVKGRFTISRDNAKSTVYLQMNSL PEDTATYYC STTAHCSGGECC ....GSFQMHYWGKGTLVTISS Ca„00326 (SEQ ID N0:#)

EVQLVESGGGSVQAGGSLRLSCEVSELIGDTYCLGWFRQAPGKEREGVCSINTFG .NAYYADSVKGRFTISQDNAEKKVFLEM SLKPEDTATYYCAYVFTSLRCSRWLQ .. YSSPQNTYWGQGTQVTVSS Ll_01011 (SEQ ID N0:#)

... LQESGGGLVQPGGSLRLSCLVSGGTFSWTAMGWFRQAPGKEREET/ACT^ RPDEYDYWGQGTQVT ...

Ca_00337 (SEQ ID N0:#)

QVQLVESGGGSVQAGGSLRLSCAVSGDTGTgyCMGWFREAPGKEREEIVALYTGGGVTWYADSVRGRFAASQDNAKNTVSLQMNSLESEDTAMYYCAAADSCVTFARVQGGTLDYSSIPYWGKGTLVTVSS Ll„OO430 (SEQ ID N0:#)

QVQLQESGGGLVQAGGSLRLSC iSGRTFSWAVGWFRQAPGKEFJ-FVGYFGTRG!GRTYYADSVKGR^ SSGTYDYWGQGTQVTVSS

Ca_00193 (SEQ ID NOi#)

DVQLVESGGGSVQAGGALRLSCQASEFTYS ... IAWFRQAPGKEREGVATGYRGVDITYYADSVKGRFTVSRDDTKNTMYLQMNSLIPEDTAIYFCAARSTYHSLR D RPGAYHAWGQGTQVTVSS

At_00003 (SEQ ID N0:#)

QVQLQESGGGLVQAGGSLRLSCAVSGRASSgyGMGWFRQVPGKEREFVAAIRWSGKETWYK^ SPVGFDYWGQGTQVTVSS

Ca„00305 (SEQ ID N0:#)

AVQLVESGGGSVQAGGSLRLSCASSGFTVSDYSMAWFRQVPGQEREWAHINADTGTTFYADSVKGRFTISLDNAKAAVYLQMNSLNTDDTAVYYCAAETAPRYYNDAV..WPSDSDFSFWGQGIQVTVSS At_00029 (SEQ ID N0:#)

QVQLQESGGGLVQAGGNLSLEXZAASGRtyGGYGMGWFRQVPGKERELVAAIRWSGTSTYYADSVKGF-FTISRDNVKi!MVYL SPVGFAYWGCGTQVTVSS

At_QQQ19 (SEQ ID N0:#)

^QV^QL^QESGGGLV^QAGGSLRLSCAASGRATSgYGMGWFR^QVPGKEREFVAAIRWSGKET YKDSVKGRFTISRDNAKTTVYLQ LPVGFDYWGQGTQVNVSS

L1_Q0111 (SEQ ID N0:#)

QVQLQEFG!GGSVQPGGSLRFSCAASGfsLDYYAIGWFRQAPGKEREGVACIRAIXKGTYYADSAKGRF^

At„00012 (SEQ ID N0:#)

QVQLQESGGGLVQAGGSLRLSCAASGRATSgyCMGWRQVPGKEREFVAAIRWSGKETWYVDSVKGRFTI^ PDGFVY GQGTQVTVSS

At_00026 (SEQ ID N0:#)

QVQLQESGGGLVQAGGSLRLSCAASGRtyGGYG GVffRQVPGKERELVAAIRWSGTSTYYADSVKGR^^ SPVGFAYWGQRTQVVSS

Ca„00306 (SEQ ID N0:#)

DVQLVESGGGSVQPGGSLRLSCEVSGDTYTG CLGWFRQAPGKERVMVATINIKHGRPYYADSVKGRFTISQDSATTT. YLQMESLQPEDTAMYFCAAIQPTYGIDFFGSCPSESADFDY GQGTQVTVSS Ca„00065 (SEQ ID N0:#)

OTQLVESGGGSVQAGGSLRLSCAVSGYTSSMYCMGWFRQAPGKEREGVAAINSGSGKTYYADFVKGRFTISQ

Ca_00352 (SEQ ID N0:#)

AVQLVESGGGSAQAGGSLRLSCWSGRTDSRDWGWFRQGPGKEREGVACIATETGNTAYTDSVKGRFTIAQDK^^ .. , PVEGAYNYWGQGTQVTVSS

L1_01Q23 (SEQ ID N0:#)

...LQESGGGLVQAGGSLGLSCAASGRTFSDYAMGWFRQAPGKE EFVSGI ...GGSTYYADSVKGE^TISRDNA HTVYLQMNRLKPEDTGVYYCASSSWWATAGTGYR...APSWYDYWGQGTQV ... At_00Q13 (SEQ ID M0:#)

QVQLQESGGGNVTAGGSLRLSCAASGRATSgYGMGWFRQVPGKEREFVAAIRWSGKETTCFUJSVKGRFTISRD LPVGFDYWGQGTQVTVSS

Ca_0Q198 (SEQ ID N0:#)

QVQLVESGGGSVQVGESLRLSCTASGYIYS ... MAWFRQLPGKEREGVATAYYRVPSiyYSDSVKGRFAISLDDAKNTVYLQMNSLKPEDTAMYYCAARTAYNPGKWD KPEIYDAWGQGTQVTVSS

Ll_00625 (SEQ ID N0:#)

QVQLQESGGGLVQTGGSLRLSCAASGFTFDDYAIGWFRQAPGKDREGVSYISSKVGRTYYVDSVKGRFTISSDNA NTWLQmSLKPED AVYYCAAIYPETLGQVL AGTLMPRYWGQGTQVTVSP

L1_00Q16 (SEQ ID NO:#t

QVQLQESGGGLVQAGGSLRLSCATSGFTFDRYAIGWFRQAPGKEREGISCIDSVG. RTWYTDSVKGRFTISSD VGEWYDYWGQGTQVTVSS

L1_Q1QQ9 (SEQ ID M0:#)

...LQESGGRLVEAGGSLRLSCLVSGGTFSWYAMGOTRQAPGKEREFVAWSRGGGSSYYADSV GRFTISRDNAKK WLQ™SLKPEDTAVYY RPDDYDYWGQGTQVT ...

Ll_00394 (SEQ ID N0:#)

...LQESGGGLVQAGGSLRLSCAASGRTFS ...MGWFRQAPGKEREFVAAIN SDSSTYYTDSVRGEIFTISRDNA NTVYLQMDSLKPVDTAIYYCAAQGSLYDDYDGL PIKYDYWGQGTQV VSS

Ll_„01007 (SEQ ID N0:#)

...LQESGGRLVEAG!GSLRLSCAASGGTFSWYAMGWFRQAPGKEREFVAOTSRGGGSIYYADSVKGRFTISRDNAKNTW ADEYDFWGQGTQVT ...

Ll_005 6 (SEQ ID NO:#)

QVQLQESGGGLVQNGGSLRLSCAASGRAFSDYTMG FRQAPGKEREFVAAITRSAFSTNYA SVKGRFTISRDNAKNTAYLQMNSLTPEDTAVYFCAAAKVSWAT PRSSDYDYWGQGTQVTVSS

Ca_00020 (SEQ ID N0:#)

QVQLVESGGGSVQAGGSLRLSCAASGVHHCINVMS YRQVPGKGREFVSSIA DG. TTRYAGDVKGRFTISQDNAKNTVYLEMNSLKPEDTAMYYCLREPLSGGRR GSCGLNYWGQGTQVTVSS

At_00085 (SEQ ID N0:#1

QVQLVESGGGSVQAGGSLRLSCAASGVNHCINVMSWYRQVPGKGREFVSSIANDG. TTRYAGDV GRFTISQDNAKNTVYLEMNSLKPEDTAMYYCLREPLSGGRRY GSCGLNYWGQGTQVTVSS

Ca_00325 (SEQ ID N0:#)

EVQLVESGGGSVQIGGSLRLSCVGSGYTDSNYCTAWFQQAPGKVREi^AVINFGRGTTQYSDSVKGRFTISQAN LVPGGYNWWGQGTQVTVSS

Li_00997 (SEQ ID N0:#)

... LQESGGGLVQAGGSLRLSCAASGRTSSTYTMGWFRQAQGKEREFWGISRSGGRTYYADSVKGRFTISRDNAKNTVYLQMN^ ...ASPREYHYWGQGTQVT...

LI_01119 (SEQ ID N0:#)

EVQLVESGGGLVQAGGSLRLSCTASRRIFPRFSMGWFRQAPGKEREFVAAISVWGGSTIYGDSVK GNDYHYWGQGIQVTVSS

Ca_00267 (SEQ ID N0:#)

AVQLVDSGGGSVQAGGSLTLSCTGSGFSFDDSELGWYHQAPGNECELVSFIGVDG .TTYYATSVKGRFAISRDNAKNTVYLRM^

Ll_01122 (SEQ ID N0:#)

QVQLQESGGGLVQAGGSLRLSCAASGRTFSSYAMGWFRQAPGKEREFVAAISWSGDSTNYADSVKGRFTISPJ⁾NAKNTVYL

Ca_001S6 (SEQ ID N0:#)

AVQLVESGGGSVQDGGSLRLSCWSGSTYS ...VAWFRQTPGKEREGVAVIFTNHRVKYYGDSV GRFTISQD TKSTVYLQKNSLKPEDTGMYYCAA PSYQSGHWSV GNLYNFWGQGTQVTVSS

Ca_Q0Q76 (SEQ ID N0:#)

QVQLVQSGGGSVQAGGSLRLSCAASGYTACSYDMSWYRQAFGKEREFVSAIETDG . STSYADSV GRFTISQDNAKNTVYLQMNSLKPEDTAMYYCKTDGVYCSGGYRWSDYWGQGTQVTVSS

At_QQQ23 (SEQ ID N0:#)

QVQLQESGGGLVQAGGNLSLSCAASGRATSgyGMGKFRQVPGKEREFVAAIRWSGKETWY DSWGRFTISPJiNAKTTVYLQMNSLKPEDTA LPVGFDYWGQGTQVTVSS

Ll_Q10Q3 (SEQ ID N0:#)

...LQESGGRLVEAGGSLRLSCLVSGGTFSWYAMGWFRQAPGKEREFVAWSRGGGSSYYADSVKGR^ RPDDYDYWGQGTQVT...

Ll_00798 (SEQ ID N0:#)

... LQESGGGLVQAGGSLRLSCAASGSIFSINT RWYRQAPGKHRELVATITSGG . STOTADSVKGRFTISRDNAKNTVYLQMNSLKPEDTAVYYCNADGIHYYSDY ... DAPPIREYDYWGQGTQVT ... Ca_00339 (SEQ ID N0:#)

QVQLVESGGGSVQAGGSLRLSCAFSGFTY RYCLG FRQAPGKEREGVASIHSGGGAPYYADSVKGRFTISFDNAKSAVDLVIKNLQPEDTAVYTCAADWPSRGWLE NCVAFNYWGQGTQVTVSS

Ca_QQlQS (SEQ ID N0-.#)

QVQLVESGGG-WQAGQSLRLSCWSEYPFKDMCLGWFHAAPGEERECTAGIYTIX^ PARVGKYP GQGTQVTVSS

Ca_00125 (SEQ ID N0:#)

QVQLVESGGGSVESGESLTLSCTADADTiSTSMMGWFRQAPGKECELVATIRTDG . STIYADSVKDRFAISQD VKNSVYLQMNSLKPEDTAVYYCAVGNCYAGSWCCV...APRRRFNFWGQGTQVTVSS Ll_01008 (SEQ ID N0:#)

... LQQSGGRLVFJiGGSLRLSCWSGGTFSWYAMGWFRQAPGKEREFVATVSRGGGSIYYADSV^ RPDEYNYWGQGTQVT ...

QVTVSS

QVTVSS QVTVSS QVTVSS

Ll„00180 (SEQ ID N0:#)

QVQLQESGGGLVQPGGSLRLSCVAFGSTSSINAMGWYRQAPGKQRELVATITSGGRS -NSADSVKGRFTISRVNAKKTVYLQMNSLKPEDTAVYSCALATKFYFGATYS .... QDRLYDYWGQGTQVTVSS Ca_00187 (SEQ ID N0:#)

QVKLEESGGGSVQAGGSLRLSCAASGLMYREYYMGWFRQAPGKEREGVAVISAPSNSAQYANSVKGRF ISRDKAKNVMYLQMNLKPEDTAIYYCAQGRGRFRTG LDAVEFDAWGQGTQVTVSS

Ca_00248 (SEQ ID N0:#)

DVQLVESGGGLVQPGGSLRLSCAASGFTFSDFAMIWVRQAPVRGLEWVSVISTSGDRTYYADSVKGRFTISRDNANTLYLQL SLKTEDTAMYYCTKDTDE PSPTD PLYGYHYWGQGTQVTVSS

Ll„00378 (SEQ ID N0:#)

QVQLQQSGGGLVQAGGSLRLSCAASGSDFSIYDIGWYRQAFGKQRELVAAIGRGG. YTNIAASVKGRFTISRDNAKNTVNLQMOTLKPEDTAWSCAAAKRYGRGRLD DITRYDYWGQGTQVTVSS

Ll_00964 (SEQ ID N0:#)

QVQLQESGGGLVQAGDSLRLSCAASGRTFSSYAMGWFRQAPGKEREFVAAVSS . ΞΝΞPWYGDSIKGRSTIHRDNAKNTVYLYMNNLKPEDTAVYYCAADLEGERLG ESSEYDYWGQGTQVTVSS

Ll_00416 (SEQ ID N0:#)

QVQLQESGGGLVQAGGSLRVSCAASGRTIGNYHMGWFRQVPGNEREFVAGSTRIG.NRYYADSVKGRFTISRDNAKNTVYLQmSLKPEDTAW ARAYDYWGQGTRVTVSS

Ll„00998 (SEQ ID N0:#)

...LQESGGGLVQAGGSLRLSCEASGRTFSSYHMGWFRQAAGKEREFVATISSDY.NTYYADSVKGRFTISRDNAKNTVYLQTTCS .... RAHEYGYWGQGTQV ...

L1_QQ671 (SEQ ID N0:#)

QVQLQDSGGGLVQAGGSLRLSCAASGRPLSDYVIGWFRQAPGKERDFVAAITWSGCTTYYADSVKGRFT^^ DKRAYDT GQGTQVTVSS

Ll_00067 (SEQ ID NO: #)

QVQLQESGGGLVQPGGSLRLSCAATGSIYSLDAMGWYRQAPGEQRELVATWTSRGITT . YADSVKGRFTISRDNAKNTVTLQMNILKPEDTAVYYCHAWYYGDWEG... SEPVQHEYDYWGQGTQVTVSS Ca_,00-28 (SEQ ID NO:#)

QVQLVESGGESVEA∞SLRLSCVSSGDSRSGDIlgWFRQAPGKEREGVAAIGRRGGTTYYSNSVKGRFTISQDKSKN^ RRSDWNYWGQGTQVTVSS

Ca_00212 (SEQ ID NO:#)

QVQLVESGGGSVQAGGSLRLSCTASGYTNVHN£¾1AWFRMPGKQREGIATLCTEWGRTYYADS GRFOTS .... LRESFYKYWGQGTQVTVSS

Ca„00350 (SEQ ID NO:#)

QVQLVESGGASAQAGESLRLSCTASGFDFADSDMGWYRQVPGNECELASSISSAG . I PLYADSVKGRFTISRDNAKNAWLQ^SLKVGDTGMYYCAADRIRLGFPRQTPACTDLRafSD GQGTQVTVSS Ll_00113 (SEQ ID NO:#)

QVQLQEFGGGLVQAGGSLRLSCAASGDAINNYGVGWRQAPGKERERVATISWDDEITLYADSVKGRFTISSDNATNTW^ AGPNGSDYWGQGTQVTVSS

At„00017 (SEQ ID NO:#)

QVQLQESGGGLVQAGGSLRLSCAASGRATSgyGMGWFRQVPGKEREFVAAIRWSGKETWYKDSVKG LPVGFDY GQGTQVTVSS

Ca„00050 (SEQ ID NO:#)

EVQLVESGGGSVQAGDSLRLSCTTSGFTFANSVMVWYRQGSGNECKLVSSISTDG .TTYYSTSVKGRFTISRDNAKNTVYLQMNNLKAEDTAMYYCAADFQASTVGRC DGYGYAKWGQGTQVTVSS

Ll_00963 (SEQ ID NO:#)

QVQLQDSGGGLVQAGTSLRLSCVAKG... GLYGLGWFRQAPGKEREFVAAISGSGGSTYYGDSVRGRFTISRDNTKNTMYLQMNSLKPEDTAIYFCAARTQYSASDYWW.... QQGEYGY GQGTQVTVSS Ca„00144 (SEQ ID NO:#)

EVQLVESGGGSVQIGGSLRLSCAASGDDYRGFYMG FRQAPGKEREGVGSIAIdaRPSGYADSAKGRFTISYDNVKNTMYLQMSNLKAEDTGTYYCA^SDPWYSA.TSDPilDYAYYmiGO/STQVTVSS Ll_00959 (SEQ ID NO:#)

QVQLQESGGGLVQAGASLKLSCEAS.. LGGLYGMGWFRQAPGKEREFVAAISGSGGTTYYEDSVRGRFTISRDNAKSTVFLQMNNLKPEDTAVYYCAARTSYSASNAW .... QLREYGYWGQGTQVTVSS Ll_00390 {SEQ ID NO: #)

QVQLQESGGGLVQAGGSLRLSCVASGNDFSIYDIGWYRQAPGKPREFVAAIGRGG . YTNIDASVKGRFTISRDNANTVYLQDTLKPEDTAVYSCAAAKRYGSGRLD DITRYNYWGQGTQVTVSS

Ll„00369 (SEQ ID NO:#)

QVQLQESGGGLVQAGGSLRLSCVASGNDFSIYDIGWYRQAPGKPREFVAAIGRGG . YTNIDASVKGRF ISRDNAKNTVYLQMDTLKPEDTAVYSCAAAKRYGSGRLD DITRYNYWGQGTQVTVSS

Ll_Q0872 (SEQ ID N0:#)

L1_0100Q (SEQ ID N0:#)

... LQESGGFJ-VEAGGSLRLSCAVSGCTFSWYAMGWRQAPGKEREFVAW RPDEYDYWGQGTQV ...

Ca_QQ294 (SEQ ID N0:#)

AVQLVESGGGSVQAGGSLSLSCAASGDT NnyCMGWFRQAPGKGLEGVAAINSGGDSSYYADSVKGRFTISQDSAKTVFLEMNSLKPEDTAIWCAAQFARATRLTCEADWYLTSAEVYVYDYVGQGVQV L1_QQ296 (SEQ ID N0:#)

^■ QVQLQEFGGGLVQTGGFLRFSCAASGRTSTINAMGWYRQASGKQRELVARV . SCGGSTNYADSVKGRFTISRDNAKNTVYLQMNSLKPEDTAVYYCAALWAGV KYSGYDYWGQGTQVTVSS

Ll_00583 (SEQ ID N0:#)

QVQLQESGGGLVQAGGSLKLTCAASEHTFSSAAMGWFRQAPGKEREFVAAIRWSGGGTYYAPSIAGRFAISG^^ ... STTTSYDY GQGTQVTVSS

Ll_00589 (SEQ ID N0:#)

QVQLQESGGGLVQAGGSLKLTCAASEHTFSSAAMGWRQAPGKEREFVAAIRWSGGGTYYAPSLA ... STTTSYDYWGQGTQVTVSS

L1_Q02Q1 (SEQ ID N0:#)

QVQLQESGGGLVQPGGFLRFSCVASGRTFTISNIAWFRQASGKEREFASAISWSG . NAYYADA KGRFTISRDNAKNTVFLQMNSLIAEDTAVYYCAAPRSSNPFLGATFLQ . KAEGYDYWGQGTQVTVSS Ca_QQQ63 (SEQ ID N0:#)

QVQLVQSGGGSVQAGGSLRLSCAASGYSISSYCMGI^RQAPGKEREGVATIFSGGLIAyyAEFVKGRFTISQDN^

Ca_QQQ68 (SEQ ID N0:#)

QVQLVQSGGGSVQAGGSLRLSCAASGHTYCSYDMSWYRQAPGKEREFVSAIDSDG . STTYADSWGRFTISQDNAIOTTVYLQroJSLKPEDTA YYCKSGTOGGYCPIDNYWGQGTQVTVSS

Ll_00172 (SEQ ID N0:#)

QVQLQESGGGLVQAGGFLRFSCAASGVAFDDYAIGV^RQAPGKEREAVSCICGSRA TYYANSVKGRFWSSD ANTVYLQMNSLRPEDTAD ... LRDRYD5WGQGTQVTVSS

L1_Q0171 (SEQ ID N0:#)

QVQLQESGGGLVQAGGFLRFSCAASGVAFDDYAIGWFRQAPGKEREAVSCICGSRA TYYANSVKGRFWSSDKAKOT ... LRDRYDSWGQGTQVTVSS

Ll„00953 (SEQ ID N0:#)

QVQLQDSGGGLVQPGGSLRLSCATSGRTFSLNSMGWYR DPGKQRQLVAAISTGG . STYYTDSVKGRFTISRDYANTVYLQMDSLKPEDTAVYYCNLVWEYRGNYY PQNDYDYWGQGTQVTVSS

Ll_00922 (SEQ ID N0:#)

... LQESGGGLVQAGGSLRLSCAASGSTFSINAIRWYRQAPGKQRELVAAI . SIGGTTNYADSVKGRFTISRDNAKNTVYLQM SLKPEDTAVYYC APGPYSDYET GYEYDYWGQGTQVT...

Ca_003 0 (SEQ ID N0:#)

QVKLEESGGGSVQVGGSLTLSCWSGDTGSLHYIjAWFRQHSGKEREGVASINIsgGRTYYADSIKGRFTISRDPGkiTLYL M LNSEDTASYYCAAGRt

Ca_00343 (SEQ ID N0:#)

QVQLVESGGGSVQAGGSLRLSCAASGATYNLycLGWYRQAPGKEREGVAHIGVHDGSTYYADSV GRFTISQDNAQNTVYLQHNGLKPEDTAIYYCA^

Ll_01087 (SEQ ID N0:#)

...LQQSGGGLVQAGGSLRLSCAASGDTFSIYTIGWFRQAPGKEREFVaalNWSGGQTDYADSVKGRFTISSDNAKNT^ PRDEYDYWGQGTQVT...

Ca_00079 (SEQ ID N0:#)

QVQLQESGGGSVQAGGSLRLSCVASGYTYRSNYMGWFRQAPGKERERVAHINSGGGSTRIADSVKGRJTISQDNAKN^

Ll_00613 (SEQ ID N0:#)

QVQLVESGGGLVQPGGSLRLSCAASGHTFSTYVMGWRQAPGKEREWAAISQS_¾3STYYAGSV^

Ca_00218 (SEQ ID N0:#)

EVQLVESGGGSVQAGGSLRLSCAASGFTGSTYLKAWRQAPGKERECTACINSIGSLFYYADSVKGRFTISQDNAKNT^

Ll_00267 (SEQ ID N0:#)

QVQLQDFGGGLVQAGGFLRFSCTASGRTLSNYGMGWFRQAPGKERDFVATISRSAYFTl^ADSV GRFTISRDNANYMVYLQimiL HTGGYDYWGQGTQVTVSS

Ca„00 69 (SEQ ID N0:#)

QVQLVESVGDSVQAGGSLRLSCVASGFTETTCVMGVTCRQAPGKERELVSSILSDGG.TVmiDSVKGP^ ...LERAYRYWGQGTQVIVSS

Ca_00107 (SEQ ID N0:#)

AVQLVDSGGGSVQAOTSLRLSCVASGYTADgdCLGWFRQAPGKGREAVASTFTGTGATYYAASWGRFAASHDIAKOT^ .. , QETYFEYWGQGTQVTVSS

Ca„00356 (SEQ ID N0:#)

EVQLVESGGGLVQAGGSLTLSCAASGFTHSSYCIGWFRQTPGQEREIWANINSGGGSTYYADSVKGRF^ .

Ll_00415 (SEQ ID N0:#)

QVQLQESGGGTVQAGGSLRLSCAASGRTYG.. ,MGWFRQAPGKEREFVAAASWSGATAYDTESV GRFTISRE VKWTVYLQH SLKPEDTAVYYCAA IRTSLSKW ESSYDYWGQGTQVTVSA

L1_007Q9 (SEQ ID N0:#)

EVQLQASGGGLVQAGGSLRLSCAVSGYTFSSHAMGWRQTPGKDREFVSAISASGGNQYYKYFAKGRFTISRDNAKNTVD^ ....VHDYDYWGQGTQVTVSS At_00030 (SEQ ID N0:#)

QVQLQESGGGLVQAGDSLRLSCAASGRtyGGYGMGKFRQIPGKERELVAAIRWSGRNTYYADSVKGRFTISR^ SPVGFAYWGQGTQVTVSS

L1_01Q91 (SEQ ID N0:#)

... LQQSGGGLVQAGGSLRLSCAASGFSSDDYTIAWFRQAPGKEREAVSCITSSM VDTDTYDYWGQGTQVT ...

Ll_00142 (SEQ ID N0:#)

QVQLQEFGGGLVQPGGFLRFSCAASGGIFAINTMAWYRQAPGNERELVAFITAAGG. NYRDSVKGRFTISRDSAKNTLFLQMNSLKPEDTAVYYCAAERKYYYTDYE NPHKYDYWGQGTQVTVSS

Ll_01050 (SEQ ID NO:#)

... LQQSGGRLVQAGGSLSLSCATSGRAFSNYAMGWFRQAPGKEREFVAAISWSGGYTYYADSVKGRFTISRDNAK^ QYEYDYWGQGTQVT ...

At_00046 (SEQ ID NO:#)

GGGSVQAGGSLRLSCNVSGSPSSTYCLGWFRQAPGKEREGVTAINTDGSVIYAADSVKGRFTISQDTAKK VYLQMHNLQPED ATYYCAARLTEMGACDAR....WATLYNYWGRGTQVTVSS

Ca_00005 (SEQ ID NO:#)

GGGSVQAGGSLRLSCNVSGSPSSTYCLGWFRQAPGKEREGV AINTDGSVIYAADSVKGRFTISQDTAKKTVYLQMNWLQPEDTATYYCAARLTEMGACDAR....WATLYNYWGRGTQVTVSS

Ll_01051 (SEQ ID NO:#)

... LQESGGRLVQAGGSLSLSCATSGRAFSNYAMGWFRQAPGKEREFVAAISWSGGYTYYADSVKGRFTISRDNAK TVYLQMNSLKLEDTAVYYCVARDYRLWSPY QYEYDYWGQGTQVT...

Ll_01075 (SEQ ID NO:#)

... LQESGGGLVQAGGSLRLSCAASGGAFSI TMGWFRQAPGKEREFVAAITRSGGntDYAESV GRFTISGDNAKNTVYLQMNRLKPEDTAIYYCAADKTTYW I PRDEYDYWGQGTQV ...

Ca_00204 (SEQ ID NO:#)

AVQLVEEGGGSVPAGGSLRLSCAASGRTYS ...MA FRQVPDKEREGVATAYTEASITYYAASVEGRFTISLDRTKSTVYLQMNSLKPEDSGIYYCASRIVYNPGHW AQVSTYH WGQGIQVTVSE

Ll_01001 (SEQ ID NO:#)

... LQQSGGGLVQAGGSLRLSCAASGRIΞΞ .NAMAWFRQNPGKERDFVATISWNGGSTDYADSLKGRFTISRDNAKNTVYLQMNSLKPEDTAVYYCAAALGKPRGAKE DARGYDYWGQGTQVT ...

Ll_00956 (SEQ ID NO:#)

EVQLVESGGGLVQPGGSLRLSCAASGSIFGINVLGWYRQAPGKQRELVASISSGG . ETTreADSV GRFTISRDNA NTVYLQMNSL PEDTAVYYCNADIGWYDEYWGQGTQVTVSSAAASGSLEQKLISE

Ll_01015 (SEQ ID NO:#)

... LQESGGGLVQAGGSLRLSCTTSGITFSDYAMAWFRQAPGKGREFVA...RSGGSTDYSDSV GRFTISRDNAKNTVYLQMNSLRPEDTAVYYCAGDYGWARPSRHS ...VRPEMYDYWGQGTQVT ... Ll_00585 (SEQ ID NO:#)

QVQLQESGGGLVQAGGSLKLTCAASEHTFSSAAMGWFRQAPGKEREFVAAIRWSGGGTYYAPELAGRFAISGNHANLVYLEMNNLKPEDTAAYYCAAMLRRFFTASYY... STTTSYDYWGQGTQVTVSS Ll_00587 ( SEQ ID NO:#)

QVQLQESGGGLVQAGGSLKLTCAASEHTFSSAAMGWFRQAPGKEREFVAAIRWSGGGTYYAPSIAGRFAI^ .. STTTSYDYWGQGTQVTVSS

Ll_00594 (SEQ ID NO:#)

QVQLQESGGGLVQAGGSLKLTCAASEHTFSSAAMGWFRQAPGKEREFVAAIRWSGGGTYYAPSLAGRFAISGNHAKNLVYLEMNNLKPEDTAAYYCAAMLRRFFTASYY... STTTSYDYWGQGTQVTVSS Ca_00322 (SEQ ID NO:#)

QVQLVESGGGSVQASGSLRLSCDASEYTFSENIMAWFRQAPGKECELVARMTSDGD . TFYG SV GRAAISRDDADNTVHLQMNSLKPEDTAVYYCVARVFVPPIN CPADFGYWGQGTQVTVSS

Ll_00637 (SEQ ID NO:#)

QVQLQESGGGLAQAGGSLRLSCAASGRTFSDY.MAWFRQAPGKEREFVAAIDYXG .. TKYRDSEKGRFTLSGDNAKNTVYLQMNNLIPEDTDAYSCAADPDLRLYYT RTDSYAY GQGTQVTVSS

Ll_00590 (SEQ ID NO:#)

QVQLQESGGGLVQAGGSLKLSCAASEHTFSSAAMGWFRQAPGKEREFVAAIRWSGGGTYYAPSI-AGRFAISGNHA l^VYLE N^ ... PTTTSYDYWGQGTQVTVSL

Ll_01046 (SEQ ID NO:#)

... LQESGGGLVQAGGSLSLSCATSGGASSEYALGWFRQAPGKEREFVAAISWSGGYTYYADSVKGRFTISRDN^ QYEYDYWGQGTQVT...

Ll_01049 (SEQ ID NO:#)

...LQQSGGGLVQAEGSLSLSCATSGRAFSSYAMGWFRQAPGEEREFVAAISWSGGYTYYADSVKGRFTISRDNA NTVYLQrDJSLKPEDTAVYYCVARDYRLWSPY QYEYDYWGQGTQVT...

L1_003B9 (SEQ ID NO:#)

QVQLQESGGGLVQAGGSLRLSCAASGRTFRNYGMGIWFRQAPGKEREFVAAISWNGGSTYYADSVKGRFTISRA AKHTVFLQMNSLKPEDTAV^

Ca_00074 (SEQ ID NO:#)

QVQLQESGGGSVQAGGSLRLSCAASGYTHCNYGMSWHRQAPGKEREFVSAIDSDG. STSYADSVKGRFTISQDNAKHTVYLQMDSLKPEDTAMYYCKRILLCGDSDYATHWGQGTQVTVSS

Ll_01086 (SEQ ID NO:#)

...LQESGGGLVQAGGSLRLSCAASGDTESIWMAWFRQAPGKEREFVAAITRSGDstDYADSVKGRFTISRDNAKNTW PRDEYDYWGQGTQVT...

Ca_00277 (SEQ ID NO:#)

QVKLEESGGDSVQAGGSLRLTCAAGTYTYHSACVGWFRQAPGREREAVAAIYPSVGRSFIADSVKGRFTVSHA^ ... SWFEGPGYNYWGQGTQVTVSS

Ca_00276 (SEQ ID NO:#)

AVQLVESGGGSAQAGGSLRLACVAAELSNRQLCMAWFRQAPGKAREIVAGKKSRGIDAVYADSVKGRFTISEDSASNTWLQMDDLKPEDTAIYYCAAA AEAGYNYWGQGTQVTVSS

LI_00939 (SEQ ID NO:#)

AVQLVESGGGLVQAGGSLRLSCAASGRTFSRYAVGWFRQAPGKQRELVAVITSGG. STKYADSVKGRFTISRDYAKHTWLQJWSLKPEDTAVYYC^KHGRWATDGSYDYWGQGTPGHRLHSGR RKAL

63 Ca_00235 (SEQ ID N0:#)

EVQLVESGGGLVQPGGSLRLSCAASGFTFS!mMSWVRQAFGKGLDWVSVINSSGGST^

Ll_00490 (SEQ ID N0:#)

... LQESGGGLVQAGGSLRLSCATSGFTFDRYAIGWFRQAPGKEREGISCIDSVG . R WYTDSVKGRFTISSDNA NTVYLQMNSLKPHDTAIYYCSVDK GSSEYG VGEWYDYWGQGTQVT...

Ll_00827 {SEQ ID N0:#)

...LQESGGGLVQAGGSLRLSCATSGFTFDRYAIGWFRQAPGKEREGISCIDSVG . RT YTDSVKGRFTISSDNA NTVYLQMNSLKPEDTAIYYCSVDKWGSSEYG VGEWYDYWGQGTQVT...

Ll_,00846 (SEQ ID N0:#)

...LQESGGGLVQAGGSLRISCAASGTTLDYYAIGWRQAPGKGREGVSCISSSGGATYYVDFVKGPJ?TISFJ^ ....YYGIDYWGKGTRVT ...

Ll_00509 {SEQ ID N0:#)

...LQESGGGLVQAGGSLRISCAASGTTLDYYAIGWYRQAPGKGREGVSCISSSGGATY^VDFVKGRFTI ....YYGIDYWGKGTRVT...

Ll_011i6 { SEQ ID N0:#)

QVQLVESGGGLVQAGGSLRLSCAASGRTFSRYSMGWYRQAPGKQRELVATISSGG . TT yADSV GRFTISRDNAKRTTYLQMNSLKPEDTAVYYCADILRTOG RNSYWGQGTQVTVSTAAASGSLEQK Ll_01059 {SEQ ID N0:#)

...LQESGGGLVQTGGSLP SCTTSGRASSNYALGWFRQAPGKERE-^AAISWSGGYTYYADSVKGRFTISRDNAEOSr-VY QYEYDYWGQGTQVT ...

Ll_00700 {SEQ ID N0:#)

DVQLQASGGGLVQAGGSLRLSCAASGRTFSSVSMGOTRQAPGKEREWAAINWRGVSTYYADSVKGRFTISRDNAKNTG

Ca_,00367 (SEQ ID N0:#)

EVQLVESGGGSVQAGGSLRLSCWSGYRYSFCSMAWYRQAPGKERELVSSTIDDG . STYYADSVKGPJTISQDNA KTVYLQIilMLKPEDTAMYYCKIVVGGYCYHPYT DYWGKGTLVTISS

Ca_00282 { SEQ ID N0:#)

DVQLVESGGGLVQPGGSLRLSCAASGFtfSNYYMSWVRQAPGKGQEWVSGIYSDGSN AYADSATKGRFTISRDNAK^

Ca_OOQ58 { SEQ ID N0:#)

QVQLVQSGGGPVQAGGSLRLSCAASRYTSCNSDMSWCRQRPGKEREFVSGIDSDG . STSYADSV GRFTISRDNAK TWLQMNSLKPEDTAMYYCKRVVLCGHSDYETY GQGTQVTVSS

Ca_00331 {SEQ ID N0:#)

EVQLVESGGGSVQAGGSLKLSCSVTGFTfsKWALGWYPiTPF CEFISTIHSDG.NRWSSD

Ll_00617 { SEQ ID N0:#)

QVQLQHSGGGLVQAGGSLRLSCAASGRTFSSYVMAWFRQVPGKEREFVAGISWSG . STTYADSVKGRFTISRDNAKNTVYLQMNSLKSGDTAVYYCAAQDSRRSPAGGSVDFGSWGQGTQVTVSS

Ca_00312 (SEQ ID N0;#)

QVQLVESGGGSVQAGGSLTLSCTASGFTFADADIGWYKESPGKQCELVSSIGSDG. RKWYEEHVKGRFTISFJDNAENTVYLQMNNLRPEDTATYACAGFPYGGSLFVC RDDRYSVWGQGTQVTVSS

Ll_00603 (SEQ ID N0:#)

QVQLVESGGGLVQPGGSLRLSCAASGSIFSINAMGWFRQAPGKQRELVAAIASGG. STWYADSVKGRFTISRDNA^

Ll_01094 (SEQ ID N0:#)

...LQESGGGLVQIGASLRLSCAASGGTDSIYTMGWFRQAFGKEREFVAAMTRsgALTDYTDSVKGFJTTISF^ PRDEYDYWGQGTQVT ...

Ll_01089 (SEQ ID N0:#}

... LQESGGGLVQIGASLPiSCAASGGTDSIYT GWFRQAPGKEREFVAAMTRsgALTDYTDSVKGR^ PRDEYDYWGQGTQVT...

Ll_00980 (SEQ ID N0:#)

...LQESGGGLVQAGGSLRLSCAASGSVASINVMGWFRQVPGF^REFVGAIGWSSGNTYYHHFVEGRFWSFJJFJ^ .RRPSDYVYWGQGTQVT...

Ll_01016 (SEQ ID N0:#)

... LQESGGGLVPPGGSLRLSCAASERTFSDYSMAWFRQAPGKDSEFVGVISWGGG . P- GDSVKGRFTISRDNAKNRMYLQMNSLKPEDTATYYCAATLDWGTIARMS ....RAGMYDYWGQGTQVT ... Ca_00213 (SEQ ID N0:#)

DVQLVESGGGSVQAGGSLRLSCTASTSEYI . YSMAWFRQAPGSPREGVATIYAVADIT ADSVKDRFIISRENAE ALVMSDLKAEDSAKYYCAARMAYSPDHWQ SSSAYQHWGQGTQVTVSS

Ca_00302 (SEQ ID N0:#)

DVQLVESGGGSVHVGGSLRLSCWsWSTIASTDCMGWFRQAPGKDREWATLLRGTTN^

Ll_00749 (SEQ ID N0:#)

QVQLQESGGRSVQSGGSLRLSCAASGIDVNRJJAMGWFRQAPGTEREFVAGVRWSDAYTDYADSVKGRFTISro QAAGYSYWGQGTQVTVSS

Ll_00341 (SEQ ID N0:#)

QVQLQESGGRSVQSGGSLRLSCAASGIDVNRNAMGWFRQAPGTEREFVAGVRWSDAYTDYADSWGR^ QAAGYSYWGQGTQVTVSS

Ll_00034 (SEQ ID N0:#)

QVQLQESGGRSVQSGGSLRLSCAASGIDVNRNAMGWFRQAPGTEREFVAGVRWSDAYTDYADS™ QAAGYSYWGQGTQVTVSS

Ll_00973 (SEQ ID N0:#)

.. ,LQQSGGGLVQNGDSLRLSCAASGRSFRTYAMGWFRQAPGKEREIVGALS SDSGTYYADSWGRFTISRDNAKOTLYLQroSLKPEDTAVYYCAADPSYARIVG SGRYIIEYWGQGTQVT ...

65



Ca_00229 (SEQ ID N0:#)

AV_QLVESGGGLVQPGGSLRLSCAASGFTFSSYAMSVA/RC^PGKGLERVSAINPDGKYTTTYPDSVKGRFTISIUDNAKNTLYLQ

Ll_00308 (SEQ ID N0;#)

QVQLQQSGGGLAQAGGFLRFSCAASGSIAIIDSMGWYRQAPGKQRELVAAI . TFGGTTTYADSVKGRF ISR-3NAK TVHLQM SLKPEDTGVYYCAADGYSSGFGTDGGSWGQGTQVTVFL

Ca_00255 (SEQ ID N0:#)

DVQLVESGGGLVQPGGSLRLSCAASGFTFSSY MYWTOQAPGKGLEOTSSINSDGGSTSYPDSVKGRFTISRDNAKWTLYLQMNTLKQEDTAVYY

Ll_00676 (SEQ ID N0:#)

QVQLVESGGGLVQAGDSLRLSCTLSGRTYTTYTMCiWYRQAPGKQRELVTSISRDD. RTOYGDSVKGRFTISRDNAKMTW

Ca_00102 (SEQ ID N0:#}

EVQLVESGGGSVQAGGSLRLSCTASGFTFDDSEMAWYQQAPG ECELVATISSSGG . TSYAESVKGRFIISRDNAKKTWLQMNSLEPEDTAMYYCATQDCSGAYCY MDPNPCGGQGTQVTVSS

Ll_00351 (SEQ ID N0:#}

QVQLQESGGGLVQAGDSLRLSCEASGPTFSRYAFGWFRQTPGQEREFVGAMTWRGGLTSVADSVKGRFTIFRDMTRNMMWLQMNDLKAGDSAV^ YSEDSYDYWGQGTQVTVSS

Ca_0O391 (SEQ ID N0:#}

QVQLVESGGGSVQAGGSLRLSCAASGYTYSSNCMGWFRQAPGKEREGVAAIYTG∞STYYADSVKGRFTISQDNAKNTWL

Ll_00424 (SEQ ID M0:#}

QVQLQESGGGLVQPGGSLPJ_JSCAASGFILNDYAIGWFRQAPGKEREGLICISSSDGTRYYADSVKGRFTISRDNAKNTWLGMNSVKPED

L1_QQQ71 (SEQ ID N0:#!

QVQLQESGGGLVQPGGSLRLSCAASGSIFSRNAMAWYRQAPGKQRELVAAIAVGGGTT . YSGSVKGRFTISRDNAK TVYLQMNSLKPEDTAVYYC ADAPAGYYPKAYWGQGTQVTVSS

Ll_00396 (SEQ ID N0:#}

QVQLQESGGGLVQPGGSLRLSC&ASGRIFRIDVMGWYRQTPGKERELVATITSGG.NTDY

Ll_00706 (SEQ ID N0:#!

DVQLQASGGGLVQAGASLRLSCAASGRSFNHYIMGWLRQAFGKERDWASIDWNSGRTNYADSVKGRFTISRDNAKNTVYLQMNSLKSEDTAIYYCAAAAAASTLVGGSYDYWGQGTQVTVSS

Ll_00945 (SEQ ID N0:#}

QVQLVESGGGLVQAGGSLRVSCAASGRAFSVRDMGWFRQAPGKEREFVAAISWNGRNITYADSVKGRFTISRDNAKNTLYLQMNSLKLEDSAVYYCAVRGASETEYSYWGQGIQVTVSSAAASGSLEQKLI

Ll_009 3 (SEQ ID N0:#}

QVQLVESGGGLVQAGGSLRVSCAASGRAFSVRDMGWFRQAPGKEREFVAAISWNGRNITYADSVKGRFTISRDNAKNTLYLQMNSLKLEDSAVYYCAVRGASETEYSYWGQGIQVTVSSAAASGSLEQKLI

Ll_00942 (SEQ ID N0:#)

QVQLVESGGGLVQAGGSLRVSCAASGRAFSVRDMGWFRQAPGKEREFVAAISWNGRNITYADSVKGRFTISRDNAKNTLYLQMMSLKLEDSAVYYCAVRGASETEYSYWGQGIQVTVSSAAASGSLEQKLI

Ll_00191 (SEQ ID N0:#)

QVQLQEFGGGLVQAGGFLRISCAASGRTFIWLRMGWFRQAPGKEREFVAVIIRSGSST-TGDSVC^RFTISRI^

Ll_00179 (SEQ ID N0:#!

QVQLQESGGGLVQSGGSLRLSCAASGTIC¾TYAMAWFRQAPGKEREFVAVIPGSCTATLYADSVKGRFTISRDNA^

L1_QQ972 (SEQ ID N0:#)

...LQQSGGGSVQAGGSLRLSCAASGRTASSWAIHWFRQAPGKEREFVAAISSSGDYTYYADSVKGRFTISRIMTKVTVYLQMNSL

Ll„00408 (SEQ ID N0:#}

QVQLQESGIGGFMQAGGSLRLSCAASGRPFSINAKGW-TIQAPGKQRELVASINRGG. TTNYADSVKGRFTISRIJNAKNTVYLQt-NS

Ca_00061 (SEQ ID N0:#)

EVQLVESGGGLVQPGESLRLSCAASGFTFSWAYMSWFRQAPGKELE SGINSDGSNTYYKDSLKGRFTISRmAIOTTLYLQMNSLKSEDTALYYCN

Ll_00028 (SEQ ID N0:#)

QVQLQESGGGLVQPGGSLRISCVASGFTFSSYYMGWVRQAPGKGLEOTSSVYIFGGSTYYADSVKGRFTISRDDAKNTLTLQ^^

Ca_00232 (SEQ ID N0:#)

AVQLVESGGGLVQPGGSLRLSCAASGFTFSDLGHSWVRQATGKGFQWVAGINSYGGSTWADSVKGRFTISRIMAICNTLYLQLNSLKSEDTAIYYCLA

Ll_00567 (SEQ ID N0:#)

QVQLQESGGGLVQAGGSLRLSCAASGSIGSIHTMGWYRQTPGKERDWATIQ.DGGSTirrM

Ll_00563 (SEQ ID N0:#)

QVQLQESGGGLVQAC¾SLRLSCAASGSIGSIHT GWYRQTPGKERDWATIQ. DGGSTNYADSVKGRFTISRDNTI^TWLQMNDL

Ll_01135 (SEQ ID N0:#)

QVQLQESGGGLVQTGDSLRLSCAVSGRTTOSYAMAWFRQAPGKEREFVAATTWRGTSIOTADPVKGRF™^

Ll_00908 (SEQ ID N0:#)

... LQESGGGLVQAGGSLRLSCAASVSTFSINAKGWYRQAPGKQRELVADISSGGG. TNYADSVKGRFTISPJOTAKNTLYLQKNSLKPEDTAVYYCNAVQQSWTRGGLRSYDYWGQGTQVT

75

77

 Ll_00049 (SEQ ID N0:#)

QVQLQESGGELVQPGGSLKLSOU^GLTFTNYSMGWFRQAPGVDREAVAAISWSGDNTYWSSVKGRFTISR^

Ll_^.00276 (SEQ ID N0:#)

_QV_QL_QESGGGLVQPGGFLRFSCAASgiSFRVQRMGWYRQAPGKQRELVATVTSAG.T NYADSVKGRFTISRDNAKNTVYLQ NSLKPED AVYYCNAGRRFMLPDYWGQGTQVVSS

Ll„00418 (SEQ ID N0:#)

QVQLQESGGDLVQPGGSLRLSCAASGFTISTFRMYWVRQAPGKGLEWVSCIDQGGESIYYADSVKGRFTT^

Ca_00399 (SEQ ID NO:#)

QVQLVESGGGSVQAGGSLRLSCAASGYTYSSCS GWYRQAPGKERELVSTIISDG. STYYADSVKGRFTISQDNAKNVYLQ NSLKPEDTA YYC

Ll_„00428 (SEQ ID N0:#)

QVKLQESGGGLVQAGESLRLSCAASGRSFSSDVMGWFRQAPGKEREFVAASSVOTGGVWHYSDSVKGRFTISRDIAKNTLYLQM SLKPEDTAVYYCRW

At„000S8 (SEQ ID N0:#)

DVQLVESGGGTVPAGGSLRLSCAASGNSLCTYDHS YRRAPGKGRDFVSGID DGTT . YVDSVKGRFTISQGWAKNTAYLQMDSLKPDDTAHYYC PSLRYGLPGCPIYPWGQGTQVTVSS

Ca_QQQ22 (SEQ ID N0:#)

DVQLVESGGGTVPAGGSLRLSCAASGNSLCTYDHSWYRRAPGKGRDFVSGIDNDGTTT . YVDSVKGRFTISQGNAKUTAYLQMDSLKPDDTAMYYC PSLRYGLPGCPIYPWGQGTQV VSS

Ll_00265 (SEQ ID N0:#)

QVQLQESGGGLVQPGGPLRFSCAASGSTFSKYWMYWWQAPGKGLE SAINTGGGLTDYAASV GRFTISRDNAKNTLYLQM SI^FEDTAVY^

Ll„01116 (SEQ ID N0:#)

QVKLEESGGGLVQPGGSLRLSCAASGRIFWLNAMGWYRQAPGKQRELVASITSGG. TTNYADSVKGPJT^

Ll_00074 (SEQ ID N0:#)

QVQLQESGGGLVQAGGSLRLSCAASASIFSIDVMG YRQAPGKQRELVAQI . TRGGSTNYAAFVKGRFTISRDNSKKTLYLQMMSIjKPED AVYYCRAEGIPETLGSRWGQGTQVTVSS

Ll_00383 (SEQ ID N0:#)

QVQLQQSGGGLVQAGDSLRLSCAASERTVGSSGRGWRQAPGKEREFVGVI WRTGQTTFADSVKG

Ca„00254 (SEQ ID N0:#)

EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYAMAWARQAPGKGLEWVSGIMDTAGRIIHYGDSVKGRVTISR^

Ll_00606 (SEQ ID N0:#)

AVQLVESGGGLVQPGGSLRLSCAASGSIFSI VMGWYRQAPGKQRELVAAI .TRGGNTNYADSVKGPJTISRDNAENTVYLQM SLRSEDTAVYYCKGIKIRAGKRSTY GQGTQVTVSTAAASGSLEQKL L1„0073Q (SEQ ID N0:#)

EVQLQASGGGLVQAGGSLRLSCAASGRTFSSYGMGWFRQAPGKEREFVGAISWGAGTPYYADSVKGRFT^

Ll_00768 (SEQ ID N0:#)

... LQESGGGSVQAGGSLRLSCTASGTIHSIVDMAWRQFPGKQRELVAARNSGG.NTTJYVDS

Ll_00562 (SEQ ID N0:#)

QVQLQESGGGLVQAGGSLRLSCAASGSIRSISYMAWYRQAPEKQRELVATITTGG . STDYEDSVKGRFTISRDSAKNVYLQKDSLKPEDTAVYYCRI YRDYNTQYFASWGQGTQVTVSS

Ll_01128 (SEQ ID N0:#)

QVQLQDSGGGLVQPGGSLRLSCAASGLSLESTWG^RQAPGKEREGISCISSGDTNTLYVDSVKGRFTISR^

Ca_00398 (SEQ ID N0;#)

EVQLVESGGGSVQAGGSLRLSCAASGYTYSSCSMGWYRQAPGKERELVSTIISDG . STYYADSVKGRFTISQDNAKNTVYLQMNSLKPEDTAMYYC

Ca„00304 (SEQ ID N0:#)

EVQLVESGGGSVQAGGSLRLTCTASELSARSRCIJWLRQAPGKEREGVAVINSDDYSANYADSVKGP^^

Ca_001 8 (SEQ ID N0:#)

EVQLVESGGGSVQAGGSLRLSCIASGYIGRYYSVGWFRQA.GREREAVASIQ GERSiyYAESAKGRFTISQEKAEntTOL

Γ,1_00027 (SEQ ID N0:#)

QVQLQESGGGLVQPGGSLRLSCAASGFTFSNYVMSWVRQAPGKGLE SVIHSSGSSTTYEDSVKGRFTISRDN^

Ll„00966 (SEQ ID N0:#)

QVQLQDSGGGLVQAGGSLRLSCAATGNMARINALNWYRQTPGKQRESVAVFSESG .TANYDDSV GRFTISRGATKDIAYLQM NLQPEDTAVYFCNAERSSGWYSGRY YRSATY GQGTQVTVSS

Ll_00621 (SEQ ID N0:#)

QVQLQDSGGGLVQAGGSLRLSCAATGNMARINALNWYRQTPGKQRESVAVFSESG .TANYDDSVKGRFTISRGATKDIAYLQM NLQPEDTAVYFCNAERSSGWYSGRY YRSATYWGQGTQVTVSS

Ca_00096 (SEQ ID N0:#)

QVQLVESGGGSVQAGGSLRLSCTASGYifRDYDLNWFRRAPGKECEFVSRIVRDGG. TAYAASAKGRFTMSRDNAKSMVYLRM NLKPEDTAVYYCAEDLSLRCNGGGQGTQVVSS

Ll_00194 (SEQ ID NO : # )

QVQLQQSGGCLVQAGGFLRLSCVASGMIFNFNDMAWmQAPGKQRESVAVITTGGT^^

81

Ll_01102 (SEQ ID N0:#)

DVQLQASGGGWQPGGSLEiLSCAASGRMYTINDMG yRQAPGNQRELVARI . SVGGT.OTKDSVKGRFTISRDNAKKTVCLQH SL PDDTAVYYCNADIRDTL DSRLYWGQGTQVTVSS ... Ll_00135 (SEQ ID NOi#)

QVQLQEFGGGLVQAGGFLRFSCTASGRIFS ...MGWYRQAPGKQRELVATISTRHTTT .YADSV GRFTVSRDNAKNTVYLQKNSLKPEDTGVYFCAADSLRIVGMNYWGKGTLVTVSS

Ll_00334 (SEQ ID N0:#)

... LQESGGGLVQAGGSLPJjSCT-ASGSTFSINPMGWYRQAPGKQREFVADIYTSTTttire^

Ll_00497 (SEQ ID N0:#)

...LQESGGGLVQAGGSLRLSCT-ASGSTFSINPMGWYRQAPGKQREFVADIYTSTTttNYGDSVKGRFT^^

Ll_00664 (SEQ ID N0:#)

QVKLEESGGGLVQPGGSLRLSCAASG. SISINAMGWYRQAPGKQREFVAAITTSG . STRYANSAKGPJTISRDNA NTWLQMDSLIPEDTAVYYCMSRAFDRTEPWRDYWGQGTQVTVSS ... Ll_00234 (SEQ ID N0:#)

QVQLQEFGGGLVQAGGFLRFSCTLASVRTFNYY^WFRQAPEKEREFVAVIDKVGESTLYGD . L1_0Q925 (SEQ ID N0:#)

... LQQSGGGLVQAGGSLPJ^SCTASGRTFSDYIMGWFRQAPGKERERVATINWTGAITYWDAVKGRFSI

Ll_00864 (SEQ ID N0:#)

... LQASGGGLVQAGGSLRLSC^-ASGRlffSTSAMGWRQVPGKEREIVASITWSGDSTYYPJDS^^ ...

L1_^00527 (SEQ ID N0:#)

... LQASGGGLVQAGGSLRLSCT-ASGRMFSTSAMGWRQVPGKEREIVASITWSGDSTYY ...

Ll_00338 (SEQ ID N0:#)

QVQLQESGGGLVQAGGSLKLSC ASRRTFSRYNMGV^RQAPGKEREFVAAIRWNIX5GADYADSVKGRFTISRD.ADGTLYLQbtNSLKPEDTAVYYCALDDIFATPGKYDYWGQGTQVTVST ... Ll_00551 (SEQ ID NO:#)

QVQLQDSGGGLVQAGGSLRLSCAASGSIGDIHTMGWYRQTPGKXRDWATIQSGG . STKYADSV GPJTISRDNTLNTVYLQMKTOLKPEDTGVYY ADVRPYRTSRYLEIWGQGTLVTVFL .. Ll_00799 {SEQ ID NO:#)

... LQESGGGLVQPGGSLRLSC&ASGSIFSRNAltAWYRQAPGKQPJSLVAAIAVGGGT

L1_0Q155 (SEQ ID NO:#)

QVQLQESGGGLVQPGGSVRLSCAASGITFRVYDMAWYRQVPGKQRELVAGI . RGGS NYADFV GPJTISRDNAKNTAYLQKNSLKPEDTAVYYCNCMP RQEY GQGTQVTVSS

Ca_00401 (SEQ ID NO:#)

EVQLVESGGGSVQAGGSLRLSCAASGYTYSSCSMGWYRQAPGKESELVSTIISDG . STYYADSV GRFTISQDNAHTVYLQM SLKPEDTAMYYC

Ll_00806 (SEQ ID NO:#)

... LQESGG!GLVQAGGSLRISCSASGFTFEDYSIGWFRQAPGKEREAVSCISTEGGYTHYADSVKDRFTISSDN^ ....

Ll_00377 (SEQ ID NO:#)

QVQLQQSGGGLVQPGGSLRLSOTASGRIFRID\MGWYRHIPGKERELVASITSSG .TANYADFVKGRFT^

Ca_00097 (SEQ ID NO:#)

AVQLVDSGGGLVQPGGSLRLSCMSGFTFSANY IWLRQAPGKGLEWSGINLGDSNTC

Ca_00256 (SEQ ID NO:#)

AVQLVESWGGLVQPGGSLRLSCATSGFTFITNDMSWVRQ^FGNGLEWLSCI^ .

Ll_00877 (SEQ ID NO:#)

... LQASGGGLVQAGGSLRLSCMSGFTFDDTGIGWFRQAPGKKREGVACISSRDGSTHYAHSVKGRFTITSDN^^ .

Ll_00540 (SEQ ID NO:#)

... LQASGGGLVQAGGSLRLSCTLASGFTFDDTGIGWFRQAPGKKREGVACISSRDGSTHYAHSVKGRFTI^ . L1_0Q718 (SEQ ID NO:#i

DVQLQASGGGSVQAGGSLRLSCAASGFTFDEHAIGWFRQAPGKGLEYVSHIDTGG . STWYAASV GRF VSRDDAHTLYLQ^SLKPEDTGLYYCARLSOGAMDY GKGTQVTVSS

Ca_00395 (SEQ ID NO;#)

DVQLVESGGGSVQAGGSLRLSCJLASEYTYSSYCMGWRQAPGKEREGVAAINSGGGSTYHADSVKGRF

Ll_00988 (SEQ ID NO:#)

... LQQSGGGLVQPGGSLRLSCAASGSIFSINSMGWYRQAPGKQRELVAYITSGG .RINYADSVKGRFTISRD , ..

Ll_01101 (SEQ ID NO:#)

DVQLQASGGGLVQAGDSLRLSCTASGRSISTYGMGRFRQTPGKEREFVAAISWSGGNEYYADSWGRFTISRDKSKNIVYLR^

Ll_01066 (SEQ ID NO:#)

... LQESGGDLVQPGGSLRLSCAASGSTLSIKAMAWYRQAPGKQREMVAAITSRG . SPKYADSVKGRFTISRDNAHTVSLQH SLKPEDTAVYYCNARTTFLTEGWRNYWGHGTQVT

84 Ca_00220 (SEQ ID N0:#)

QVQLVESGGGSVQAGGSLILSCTDSEAAYTRNCMGWFRQAPGKEHEGVAALPYGDGPTYYDDDVRGRFTISRDNAKNTLYLTMKDLEPEDTAMYYCAAKQGPFCGTWYLWRTFAAWSQGTQVTVSS

Ll_003B7 (SEQ ID N0:#)

_QV_QLQESGGGLVQPGGSLTLSCAASGRIFRIDFMGWFRQSPGKQRE VATL . TGDDIANYSDSVKGRFTISRDNSK TFYLQM SLKPDDTAVYYCNARGPYSRGSGP GQGTQVTVSS

Ll_00360 (SEQ ID N0:#)

_QV_QL_QESGGGLVQAGSSLRLSCAVSGRPFRRSSVGV^IQAPGKEREFVATLTWSG.SSEYAESVKGRFTISRDNHRDT^

Ll_00774 (SEQ ID N0:#)

...LQESGGGLVQAGGSLRLSCGASGSTFSISTMGWHRQAPGKEREYVANIGSLG. TTDYADSVKGRFTISRDNAKNTVYLQMNSLKPEDTAVYYC QDGLYGFTGKNYWGQGTQVT

Ca_00252 (SEQ ID N0:#)

DVQLVESGGGSVQAGGSLRLSCAASRYTDCKYDMSWRQVPGKERELVSQINKIGTDTYYWSVKGRFWSRDDAKHTI-N

Ca_002B9 (SEQ ID N0:#)

AVQLVESGKSVQTGGSLRLTRALSGNMGRFNSMGWIRQAPGKGREAVACIHTGGGSAWA

Ll_00947 (SEQ ID N0:#)

QVQLQESGGGLVQTGGTLRLSCTASGNTNGIHDMGWFRQAPGNQRELVATISSGG. TIRYADYVKGRFAISRDNAETTVYLQiiNSLKPEDTAVYYCYVDRAWVNYWGQGTQVTVSSEPKTPKPQPAAASGS Ca_00207 (SEQ ID N0:#)

EVPLVESGGGLVQAGGSLRLSCVASGFSISSCCtlKVireRQAPGKERELVSSITSAANTYyyADFPVGRFAISRDGAKNILYLQM SLKPEDTAVYY

Ll_00491 (SEQ ID N0:#)

... LQESGGGLVQAGGSLNLSCTAVGNIFRINAMGWYRQAPGKQRELVAQI . SRGSSTNYADSVKGRFTISRDNAKDTVTLQMNSLKPEDTAVYYCNAQGRYYGGTYDPTYDYWGQGTQVT

L1_00S28 (SEQ ID N0:#)

... LQESGGGLVQAGGSLNLSCTAVGNIFRINAMGWYRQAPGKQRELVAQI . SRGSSTNYADSVKGRFTISRDNAKDTVTLQMNSLKPEDTAVYYCNAQGRYYGGTYDPTYDYWGQGTQVT

Ll_00750 ( SEQ ID NO : # )

QVQLQESGGELVQPGGSLKLSCAASGLTFTNYSMG FRPGPGVDREAVAAIS SGDNTYYVSSVKGRFTISRDNAKNTVYLQMNSLKPQDTAVYYCAVKPDIX^OTroYWGQGTQVTVSS

Ll_00547 (SEQ ID NO:#)

QVQLQESGGGCTQPGGSLRLSCAASGFSFGDYAIGWFRQALGKEREQISCINDNGGITSYRGSLKGRSTISRDNAKNTVYLQMNSLEPEDTAV^

At_0004B (SEQ ID NO:#)

GGGSVQAGGSLRLSCasSKYMPCTYDMTWYRQAPGKEREFVSSINIDGKTT.YADSVKGRFTISQDSAKNT^

Ca_00009 (SEQ ID NO:#)

GGGSVQAGGSLRLSCasSKYMPCTYDMTWYRQAPGKEREFVSSINIDGKTT .YADSVKGRF ISQDSAKNTVYLQMNSLKPEDTAMYYCKIDSYPCHLLDVWGQGTQVTVSS

Ll_00203 (SEQ ID NO:#)

QVQLQQFGGGLVQPGGFLRLSCAASGRIFRLNAMGWYRQAPGKQRDLVARIAIGG . VTT^ADSVKGRFTISRDNAKNTVYLQMNSLKPEDTAAYYCNAKYYSGF¾rYWGQGTQVTVSS

Ca_00246 (SEQ ID NO:#)

EVQLVESGGGLVQSGGSLRLSCVASGFAFNNYAMSWGRQAPGKGLEWVSTINSAG . TTYYADSVNGRFTISRDNAKNTIYLQLNSLKPEDTGTYYCSKTGEIWGQGTQATVSSESS

Ll_00220 (SEQ ID NO:#)

QVQLQESGGGLVQPGGFLRFSCAASGIIVSRNAMAWYRQAPGKQREFVATIGS . GSMTNY DSVKGRFTISRDNAKNTVYLQMNSLKPEDTAVYYCKSIGRAAWGQGTQVTVSS

L1_00S59 (SEQ ID NO:#)

...LQESGGGSVQAGGSLRLSCAASGGAFSGLVMGWFRQVPGKEREFVAQINRHGDTPSYADAVKGRFSISRDNAKNTVYL

Ll_00522 (SEQ ID NO:#)

...LQESGGGSVQAGGSLRLSCAASGGAFSGLVMGiWFRQVPGKE EFVAQINRHGDTPSYADAVKGRFSISRDN^

Ca_00387 (SEQ ID NO:#)

AVQLVDSGGGSVQAGGSLRLSCAASGYTFSSYAMGWFRQAPGKECELVSTIISDG. STNYADSVKGRFTISRDNAKNTVYLQMNSLKPEDTAMYYC

Ca_00253 (SEQ ID NO:#)

DVQLVESGGGLVQPGGSLTLSCAASGFTFSAYW YWVRQTPETGLE SAIYGK1X3QTVYPDSVKGRFTISK^

Ca_00136 (SEQ ID NO:#)

AVQLVESGGGSVQSGQSLRLSCTVFRYTNDNYCVAWFRQAPGKEQEAVAGLtvGGGKTWYADSVKGRFTISQDHARSAKY

Ca_„00402 (SEQ ID NO:#)

EVQLVESGGGSVQAGGSLRLSCAASGYTYSSCSMG YRQAPGKERELVSTIISDG. STYYGDSVKGRFTISRDNAKSTVYLQMNSLKPEDTAMYYC

L1_OQ950 (SEQ ID NO:#)

QVQLQESGGGLVQTGGTLRLSCTASGNTNGIHDMGWFRQAPGNQRELVATISSGG. TIRYADYVKGRFAISRDNAETTVYLQMNSLKPEDTAVYYCYVDRAWVNYVfGQGTQVTVSSEPKTPKPQPQPQPQP Ll_00812 (SEQ ID NO:#)

... LQESGGGLVQTGGSLRLSCATSGRTFSSFVMGWFRQAAGTEREFVSAISERGSLIHYADFVKGRFTISRDNAENTVSLQMNSLKPEDTAVYYCAAVIGGRWPPSEYAYWGQGTQVT

87 Ll_00949 (SEQ ID N0:#)

QVQLQDSGGGLVQTGGTLRLSCTASGNTNGIHDMGWFRQAPGNQRELVATISSGG. TIRYADYVKGRFAlSRDNAETTVYLQMNSLKPEDTAVYYCYVDRAWVNYWGQGTQVTVSSEPKTPKPQPAAASGS Ll_00112 (SEQ ID N0:#)

QVQLQEFGGGLVQPGGSLRLSCVGSGFTSDTYAMGWFRQAPGKEREGVASLDFVTDKAFYAESAKGRFTISRDNAKD

L1_QQ258 (SEQ ID N0:#)

QVQLQESGGGLVQPGGFLSFSCAASGSIFLINAMGWYRQAPGKQRELVA. IITNGGT NYADSVKGRFTISRD TKNWLQDSLKPEDTAVYYCNGDHYSTQGGQLY GQGTQV VSS

LI_00836 (SEQ ID N0:#)

... LQESGGGLVQAGGSLRLSCAASGFSLDDYDIGWFRQAPGKQRELVASISFLGHTTNYADSVKGRFTI

LI_Q0499 (SEQ ID N0:#)

... LQESGGGLVQAGGSLRLSCAASGFSLDDYDIGVffRQAPGKQRELVASISFLGHT NYADSVKGRFT^

Ll_00484 (SEQ ID N0:#)

GDLVQAGGSLKLSCAATGFSSDDYVMG FRRAPGKDRVGVACINSKTGRTYFRPDVQGRFTISSDNAKH WLQMNSVNVDDTAVYSCAATESLIATDWC .... LSTVSYNYWGQGTQVTVSS

At_00062 (SEQ ID N0:#)

GDLVQAGGSLKLSCAATGFSSDDYVMGWFRRAPGKDRVGVACINSKTGRTYFRPDVQGRFTISSDNAKHTVYLQMNSVNVDD AVYSCAATESLIATDWC .... LSTVSYNYWGQGTQVTVSS

Ca_QQ141 (SEQ ID N0:#)

AVQLVESGGGSVQAGGSLSLSCADVGRsnSNAGICWFRQAPGKERECVGTIYLggRVTYYSDSVKDRFTISPDNAGKT^

Ll_00068 (SEQ ID N0:#)

QVQLQESGGGLVQPGGSLRLSCAASGSIGSI AHGWYRQAFGKQRELVAQF .TPGGSTEYADSVKGRFAISRDNAKTTVYLQMNSLKPEDTAVYYCYCIN SDYWGQGTQVTVSS

Ca_00396 (SEQ ID N0:#)

QVQLVESGGGSVQAGGSLKLSCAASGYIFSSCGMGWYRQAPGKERELVSTISSDG . STSYADSVKGRFTISQDNAKNTLYLQMNSLKPEDTAMYYC

Ll_00680 (SEQ ID N0:#)

QVQLVESGGGLVQPGGSLRLSCAASGFTLDYYAIGWFRQAPGKEREGVSCISSSDGSTYYADSVKGRFTISRIOTAKNTWLQMNSLKPEDTAVYYCM

Ca_00228 (SEQ ID N0:#)

QVKLEESC3DLVQPGGSLRLSCATSGFTFSNFWMSWIRQAPGKGLKWSGINTDGTITRYEDSVKGRFTISRDNAKNTLYL

Ll_00793 (SEQ ID N0:#)

... LQQSGGGLVQPGGSLRLSCAASGSITSIRSMGWYRQAPGKQREFVAHISTGG . ATNYADSWKDRFTISRDNAI^T YLQl^SLKPEDTAIYYCNALLYYSDYElffiNYWGRGTQVT

Ll_00093 (SEQ ID N0:#)

QVQLQESGGGLVQAGGSLRLSCAASGITFSNDASGWSRQAPGKQLDFIARIRSGG. TTVIADSVKGRFTISRDDAKNTVYLHMNSLKPEDTAVYFCNADSNGRTYWGQGTQVTVSS

Ll_01097 (SEQ ID N0:#)

DVQLQASGGGLVQAGDSLRLSCAASGRTSSTYAMAWFRRAPGKEREFVTAISWNGRITAYADSVKGRFTISREYAGNTVTLQMDSLKPEDTAVYYCAADYTTAVPRAHVSYDYWGQGIQVTVSS

L1_QQ731 (SEQ ID N0:#)

EVQLQASGGGLAQPGGSLRLSCTADGRTFSDIAMA FRQAPGKEREIVAAIDWNGGTTYYTTFV GRFTISRDNAKKTVYLQMTSLKPEDTAVYYCKALDITTAASYWGQGTQVTVSS

Ll_01109 (SEQ ID N0:#)

DVQLQASGGGLVQPGGSLRLSCAASGSLSRITVMGWYRQAPGKQRELVAIITSSGG . TDYADSVKGRFTISKDNAKALMYLQMTSLRPEDTAVYYCAGKSRDSAGLSWDY GQGTQVTVSS

Ll_0Q724 (SEQ ID N0:#)

EVQLQASGGGLVQTGGSLRLSCAVSGRTFSSYGMGWFRQAPGKEREFVAAMRESGADTHYADFVRGRFTISGDNAKN^

Ll_00254 (SEQ ID N0:#)

QVQLQESGGGLVQAGGSLRFSCAAAGSTSVTYTMGVffRQAPGKEREFVFWSRSAGITRYAD-T/KGRFT^

L1_0033Q (SEQ ID N0:#)

QVQLQESGGGLVQPGDFVRLSCAASRRASSTYAVGWRQAPGKEREFVGRIHRGGGSTYYADSVKGRFTISR^

Ca_00116 (SEQ ID N0:#)

WQLTCSGGGSVQAGGSLRLSCAVSGYTVSSTCMG^RWGINERAGVAAISSGGAATVreEESV

Ll_0Q9Q6 (SEQ ID N0:#)

... LQESGGGLVQAGGSLTLSCAASGSTSSrnAKGWYRQAPGKQRELVASIN . IXMSTTYADSVKGRFTISRDNAKNTMSLI-NSLQPEDTAVYYC ARTOFFTTYVNNWGQGTQVT

Ll_00144 (SEQ ID N0:#)

QVQLQESGGGLVQAGGSLRFSCAALGSRFSGKAMAWYRQAPGKPRDLVARIENDG . STNYAVSVKGRFTISKDDAKNTVOTjQilNSLKPEOTAVYYC ANELIAPG ILYLFATWGQGTLVTVSS

L1_QQ681 (SEQ ID N0:#)

QVQLVF-SGGGLVQPGGSLRLSCAASGFTLDDYAIGVffRQAPGKEREGVSCISSSDGSTYYADSVKGRFTISRDNANTLYLQMNSLKPEDTAVYYCAA

Ll_00246 (SEQ ID N0:#)

QVQLQESGGGLVQPGGFLRFSCAASGFTFGNQYIlJWVRQASGKGPEWAAIia_GSDRTYYADSVXGRFTISSDNAAN^

Ca_00202 (SEQ ID N0:#)

QVQLVESGGGLVQPGGSLRLSCEASGFIFSAYPMTWYRLAPGKGLESVSRITSDGFJSTVreSDSVKG

L1_0Q264 (SEQ ID N0:#)

QVQLQESGGGLVQPGGFLKFSCAASSSSISFDAMGWYRPAPGKQRELVATISSDG.TTDYSDSTO

Ll_00114 (SEQ ID N0:#)

QVQLQEFGGGLVQPGGFLTFSCAASGRTFSDYDMGWFRDVPGKDREFVAAIDGNGQTTVYVES™

Ll_00660 (SEQ ID DTO:#)

QVQLQESGGGLVQPGGSLRLSCAAFGFTLDQHAIGWFRQSPGNERFJWSCINAMXSAIYYADSVK^

Ll_00720 (SEQ ID N0:#)

EVQLQASGGGLVQPGGSLRVSCAASGFTFS HQMSWVRQAPGKGLEYVSHIDTGG. STWYAASVKGPJTVS DDAKNTLYLQMNSLKPEDTCLYYCARLSQGAMDYWG GTQVTVSS

Ll_00856 (SEQ ID DTO:#)

... LQQSGGGSVQPGGSLFJ-SCAASGFSFGDYAVGWFRQAPGKEREQVSCINDNGGITSYVGSAKGRFTISR^

Ll_00519 (SEQ ID N0:#)

... LQQSGGGSVQPGGSLRLSCAASGFSFGDYAVGWFRQAPGKEREQVSCINDNGGITSWGSAGRFTISRDNAKK^

Ll_00198 (SEQ ID N0:#)

QVQLQESGGGLVQAGGFLRFSCVAPGSIFSGNDMGWYRRAPGKQRDLVA . VF NGGSTNYADSVKGRFTISRDYAKMTMYLQMHSLKPED AVYYCVDLLAVDADPYRYFEWGQGTLVVSS

Ll_00222 (SEQ ID NO:#i

QVQLQEFGGGLVQPGGFLRFSCAASGSIFRANAMGWYRQAPGKERELVARI SGG. IP YADSWGRFTISFDNAENTVYLQM SLKPEDTAVYYCKGI KKTWFPGMDYWGKETLVTVSE

Ll_00454 (SEQ ID N0;#}

GGLVQPGGSLRLSCAASGFTFSSYDMSWTOQAPGKGLEWVSGIYSDGTTTYDGDSVKGFJTISFIN^

LL.00732 (SEQ ID N0:#1

DVQLQASGGGSVQAGASLRLSCATSGQTLNTYVM(_WFRQAPGKEREFVAAINWRD S^

Ll_00251 (SEQ ID N0:#)

QVQLQEFGGGLVQAGGFLRFSCAASGSIASINYMGWFRQAPGNQREFVARITSGG . T YRDSVKGRFTISRDSAKTiTVELQMNSLKPEDTAVYYCTFIAAGGENN GQGTQVTVSS

Ll_00294 (SEQ ID N0:#)

QVQLQESGGGLVQAGGFLRFSCAASGITFSITTMGWYRQAPGNQRELVATITGCG. ST YADPVKGRFTISRDNAENTVYLQOTSLKPEDTAVYYCKPAAWLNYWGQG QVTVSS

Ll_00138 (SEQ ID N0:#)

QVQLQEFGGGLVQAGGSLFJ-SCAASGRIFSIRPMGWYRQAPGKERELVALISSGG. I TTYADSVKGRPriARDGTGWrVYLQ^SLKPEDTSVYYCTRAAVG WWGQG QVTVSS

Ca_00409 (SEQ ID DTO:#)

DVQLVESGGGLVQAGGSLRLSCAASGFTYSSCCMSWYRQAPGKERELVSSISSDG. STYYADSTOGRFTISKDNAKNTLYLQMNSLKPEDTAMYYC

L1_0Q797 (SEQ ID N0:#)

... LQQSGGGLVQAGGSLRLSCAASGIIFSVRAHGWYRQAPGKQRELVAVILRGG. STNYADSVKGRFTISRDRAKNrAWQ^SLKPDDTAVYYOTANVLTRALVSGDYWGQGTQVT

Ll_00880 (SEQ ID N0:#)

... LQESGGGLVQAGGSLKLSCTASRRTFSRYI^GWRQAPGKEREFVAAIRWIIGGADYADSVKGRFTISRD . AIXrLY 3MNSLKPEDTAVYYCALDDIFATPG YDYWGQGTQVTVSTAHHSEDPSS . L1_^00327 (SEQ ID N0:#)

QVQLQESGGGLVQAGGFLRFSCAASGRTFSRYTMGWFRQAPGNERKFVAAVSTSG.NTHYTGSVKGRFTIFRQNAK TVYLQMSNLKPEDTAVYYCAARFGGM WKYWGQGIQVTVSP

Ll_00328 (SEQ ID N0:#)

QVQLQESGGGLVQAGGFLRFSCAASGRTFSRYTTCGWFRQAPGNER FVAAVSTSG.NTHYTGSVKGRFTI^

Ll_00143 (SEQ ID N0:#)

QVQLQEFGGGLVQAGGFLRFSCAFSGNTRSVN GWYRQAPGKQRDLVAMI RDG. STNFADSVKDRFTISRDIANSTLYLQMNSLKPEDTAVYYCHAEGLISGISPFDSWGRGTQVTVSS

Ca_00156 (SEQ ID N0:#)

AVQLVESGGGSVQPGESLRLSCLVSGYAAT ... LGWFRQAPGKECELVATISRDG. FTKYGDSVKGRFTVSRDNNKN MYLI-¾TSLKPDDAGVYYCAHAFL™GI5DCPDDRAGA GQGTQVTVSS

Ll_01105 (SEQ ID DTO:#)

DVQLQASGGGKVQAGDSLRLSCAASERID TYAVAWFRQAPGKEREFVAAIH RG. STWANSVKDRFSVSRDNAKNTWIflMNSLKPEDTAW

Ll_00748 (SEQ ID N0:#)

QVQLQESGGGLVQAGGSLTLSCAASGPMSSSAVMSWFRQAPGKEQEi^ARIRWSGGTSYYANSVEGRFTISF^

L1_001Q5 (SEQ ID N0:#)

QVQLQESGGGLVQAGGSLTLSCAASGPMSSSAVMSVffRQAPGKEQEI^ARIRWSGGTSYYANEWE^

Ll_00769 (SEQ ID N0:#)

... LQESGGGLVQAGGSLRLSCAASGSIFSVNAMGWYRAAPGRQRELVASITSGG. STNYADSVKGRFTISFINAKNMY 3MNDLKPEDTAVYYCTNCRVGNWRLEGSWGQGTQVT

-1 L1_0Q634 (SEQ ID N0:#)

DVQLVESGGGLVQPGGSLRLSCRPSISIFSIDLMGWYRQAPGKEREWATI . TKGGRTSYVDAVKGEiFTISTDNSKNTVFLQMNRLTPEDTAVYYCAA ISVGGDHSLEMGQGTQVTVSS

Ll_00063 (SEQ ID N0:#)

QVQLQESGGGLVQPGGSLKLSCAVFGSIVRGNTMAWYRQAPGKERELVAH .YTSGGSIDYSDSV GRFTISRDSLKNTVCLQ1IHSLKPEDTAVYYCNADVWYGST RNYWGQGTQVTVSS

L1_0063I (SEQ ID N0:#)

QVQLQESGGGLVQPGGSLRLSCTVSGRFFSPYPMG YRQTPGKQRELVADITYNGRT . YADSVKGRFTISRIOTA KIMYLQMNNLKPEDTAVYYCEiRPGYLLS GQGTQV VSS

L1_0Q682 (SEQ ID N0:#)

QVQLVESGGGLVQAGGSLRLSGAASGDTICISAMGWYRQAPGKERELVAAITSGG. STNYADSVKGRFTISRDNAK VYLQMNSLKPEDTAVYYCNA

LI_01068 (SEQ ID N0:#)

... LQXSGGGLVQAGGSLRLSCAASGGSFSI TMAWFRRAPGKDREFVAAISRSGGstDYAESVKGRFTISRDSAENTVFLRMNSIJKPEDTAIYYCAADKTTFWNIPRDEYDYWGQGTLVT .... Ll_00367 (SEQ ID NO:#i

QVQLQQSGGGLAQTGGSLRLSCVASGSISSINAMGWYRQAPGKQRELVAAITSGG. SARIKDSLKGRFTISRDN^

LI_00270 (SEQ ID N0:#}

QVQLQEFGGGLVQPGGFLRFSCAASGSIFSGNAMGWHRQVPGKQRELVGAITTDGC . T^AASVKGRFTISRDNAKHWYLQllWSLKPEDTAVYY<_mD^^

Ca_00008 (SEQ ID N0:#}

GGGSVQAGGSLRLSGAASDYTITDYC AWFRQAPGKERELVAAivVDTRLTDYADSVKGRFTISQGOT .

At_QQQ43 (SEQ ID N0:#}

GGGSVQAGGSLRLSCAASDYTITDYCMAWFRQAPGKERELVAAivVDTRLTDYADSV GR^

Ca_00244 (SEQ ID N0:#}

AVQLVESGGGLVQPGGSLRLSCAGSGFMFSGAAM WVRQAPG GFEWVSRINVDG. TTYYAESVKGRFTISRENA KTLYLQIJISLRTEGTAMYYCSTIG WRYYNLEYWGKGTLVTISPESS.. Ca_QQ271 (SEQ ID N0:#)

QVQLVESGGGSVQAGGSLRLSCAPSGGPTCSHQ EWYRQAPGKEREFLSGIELDG . RTMYQDSVKGRFAISHDNL STVFLQMNSLKPEDTAKYYC LTARRCGFTTT LELWGQGILVNVSS.. Ll_00014 (SEQ ID N0:#)

QVQLQESGGGLVQTGGSLRLSCTASGFTFDDYAICWRQAPGKGREGVSCISKEYAKTYYTDAVEGRFTISSDDAK1CT

LI_0Q298 (SEQ ID N0:#)

QVQLQQSGGGLVQPGGFLRFSCAASGSIFSINAMGWYRQAPGMCRELVAVITSIXJ.STNYGDSATGR

LI_00059 (SEQ ID N0:#)

QVQLQESGGGLVQAGGSLRLSCTASGRTLENYRVAllWRQFPGKERELIAVINWSGDRRYYTEAV^ . Ll_00207 (SEQ ID N0:#)

QVQLQQSGGGLVQAGGFLRLSCAASGNITRINAMGWYRQAPGKQREFLA. IITIGGTI YADSVKGRFTIARGNAENSVYLQM SLEPEDTPVYYCAAVISGSDYGPHDSWGQGTQVTVSS .... LI_00948 (SEQ ID N0:#)

QVQLQETGGGLVQTGGTLRLSC ASGN NGIHDMGWFRQAPGNQRELVATISSGG. TIRYADYVKGRFAISRDNAET WLQMSTSLKPED AVYYCYVDRAlimiWGQGTQVlVSSEPKTP . . . . Ll_00106 ( SEQ ID N0:#)

QVQLQESGGGLVQAGGSLRLSCAASGPTLGSYVMGVffRQAPGKEREYVGSITVreGGINTYYA ..

Ll_00622 (SEQ ID N0:#)

QVQLQESGGGLVQPGGSLRLSCAASGSIATIGD GWYRQVQGTQRELVAVI .RRGGTIHYADFVKGRFIISKDNAKN IDLQ1IHSLKPED AVYYCAAERIKVGGRMLIYWGQGTLVTVSS .... Ll_00001 (SEQ ID N0:#)

QVQLQESGGGLVQAGGSLRLSCVAQGR WWDLDMGWFRQAPGNQREFLAAITSGG.TPHYADS...RFTLSRDHAKNLVYLQM SLKPEDTARYYCAAGAYSTRPEAYRY GQGTQVTVSS .... Ll_00035 (SEQ ID N0:#)

QVQLQESGGGLVQAGGSLRLSCAASGSSLSNYTMGWFRHFPGKDREFVAWKWSGGRTHYTESWGRFTTGRDTAJ-fTIVYLQlIHRLKPEDTAIYYCAAASRWSWNPTSDDYIYWGQGAQVTVSS . Ll_00762 (SEQ ID N0:#)

... LQESGGGLVQAGGSLRLSCAASASIFSIDVKGWYRQAPGKQRELVAQI .TRGGS NYAAFVKGRFTISRDNSKKTLYLQMNSLKPEDTAVYYCRAEGIPETLGSR GQGTQVT

Ll_00052 (SEQ ID N0:#)

QVQLQQSGGGLVHAGGSMRLSOUVPGRTFSSYAMA IRQAAGKEREFVAGV-^GGLTWGDFVEGRFTISRDSTKNTW .... LI_OQ107 (SEQ ID N0:#)

QVQLQESGGGLVQTCASLRLSCVASGRTLSRYT AWIRQAPGKGREFIGRITWTDGSTYY ..

LI_01029 (SEQ ID N0:#)

... LQESGGGLVQPGGSLRLSCVASGTIFSINaiGWYRQAPGKERELVAAITADGTSA. YEDSVKGRFIISRDDAKKMVYLQMNSLKPEDTAVYYC GLRASNAGWEPRFGT GQGTQVT

Ca_00423 (SEQ ID N0:#)

EVQLVESGGGLVQPGGSLRLSCMSGFTFSSYWMYWVRQAPGKGPE VSTINSGGGSTYYADSVKGRFTISRDNA ^

92

LI_00496 (SEQ ID N0:#)

... LQESGGGWVQPGGSLTLSCDASGSGFRG VKAWYRQTPGKRRELVASITSDD .RTNYADSTOGFJ?TISRDNAKNMMHLQMNSLKPEDTAVYYCHVDTGGPYGSRNPYWGRGTQVT

Ll_0Q656 (SEQ ID N0:#)

QVQLQDSGGGLVCAGGSLRLSCAVSGRTDSNYVMGWSRQAPGKGREFIAAIHWSEGGTHYADSVKDRFTIPRDSAi^

Ll_00081 (SEQ ID N0:#)

QVQLQESGGDLVQAGGSLRLSCVASGIIFRIHHMGWYRQAPGKQRELVARISYDD . IISYADSVKGRFTISRDNAKDTVYLQMNGLKPGDTGVYSCHAHNKLAPPGKLEYWGQGTLVTVSS

Ll_00080 (SEQ ID N0:#)

QVQLQESGGDLVQAGGSLRLSCVASGIIFRIHHMGWYRQAPGKQRELVARISYDD . IISYADSVKGRFTISRDNAKDTVYLQMNGLKPGDTGVYSCHAH KLAPPGKLEY GQGTLVTVSS

Ll_00821 (SEQ ID N0:#)

... LQQSGGGLVQAGGSLRLSCVASGIIASDSAMAWYRQAFGKGREYVAHI .TAGGLS YEDFVNGPJTISRDNARNTMYLQM SLKPED AVYYCNVGQSYIRSYY PGDY GQGTQVT

Ca_^001Q5 (SEQ ID N0:#)

AVQLVESGGGLVHPGESLKLSCftASGFTFGTKFMSWWQ^PGKDLEWSIINSGGENTWYADSV GRFIIYRDNA AMLYLHMHALKTED

Ll_00623 (SEQ ID N0:#)

QVQLQDSGCKLVQAGDSLRLSCTASGSIATIGDMGWYRQVQGTQRELVAVI .RRGGTIHYADFVKGPJTISGDNAiOTTVLLQ^SLKPEDTAVYYCAAETAPHMDIYWGQGTLV VSS

LI_00678 (SEQ ID N0:#)

QVQLVESGGGSVQPGGSMKLSCAFPRSIFSNNARAWYRQAPGKQRELVARISSGG . ITNYADSVKGRFTISRDNAiOTTIHLQMNKLKPEDTATYYCNVRRWTKDYWGQ^IQVTVSS

Ca_0038B (SEQ ID N0:#)

EVQLVESGGGLVQPGGSLRLSCAASGYTFSSYAMGWRQPPGKECELVSTIISDG.STNYADSVKGFJFTFS

Ll_00079 (SEQ ID N0:#)

QVQLQESGGGWQPGGSLRLSCAASEMIFDF DMGVWRQAPGKERELVASISRFG.RTNYIDSVKGR^^

Ll_01065 (SEQ ID N0:#)

... LQESGGGLVQAGGSLRLSCAASASTFVINPMGWYRQAPGKQRELVAGITFNG .AimADSV GRFTISl^AiOTTWL^SLKPEDTAVYYi-WAVVWGSESYDS GQGTQVT

LI_00329 (SEQ ID N0:#)

QVQLQESGGGLVQPGPFLNVSCVVSGGIFSDYTLGWFRQAPGKERKFVAAVSSGG . STHYTGSVKGRFTISRDNAA TMYLQMSSLKPDDTAVYYCNAIVPPTRTFCGRTYWGQGTQVTVSS .... LI_00109 (SEQ ID N0:#)

QVQLQQFGEVLVQEGGSLRFSCTASGRKFKDYWGWFRQAPGKERELVASISKIEGRTAIGDSVKGRATISRDNAiOT^

Ll_00084 (SEQ ID N0:#)

QVQLQESGGGLVEAGGSLRLSCVASGNIFGVNPVSWYHQTPGKQRELVATITSDG . STTYSDSVKGPJTISRDNAKKTVDLQmSLKPEDTGVYDCRW TNRGMDYWSQGTQVTVSS

L1_0Q822 (SEQ ID N0:#)

... LQESGGGLVQPGGSLRLSCAASGSIFSMNTVSWYRQAPGNQRELVASIRGDG . R SYEDFV GRFTISRDIAKNTVYLQMNNLKPEDTAVYYCNAWISADASMMTTNYWGQGTQV

Ll_00445 (SEQ ID N0:#)

... LQESGGGLVQPGGSLRLSCAASGSIFSMNTVSWYRQAFG QRELVASIRGDG . RTSYEDFVKGRFTISRDIAKNTWLQMNNLKPEDTAVYYCNAWISADASMMTTNYWGQGTQV

Ll_00156 (SEQ ID N0:#)

QVQLQEFGGGLVQAGGSLRFSCAASGSIFRFDVmnWFRQAPGKQPELVAQI.TRGSSAYYVESVRGRFTISRDNAKNTWLQMNSLKPEDTAVYYCWPDPPLDY GQ

Ll_01073 (SEQ ID N0:#)

.. -LQESGGGLVQPGGSLRLSCVASGINSRIYRTGWYRQAPGNQRELVARI .SDGGSTNYGDSVKGRFTISRL^AKNTWLQMNNLKPEDTAVYQCCVNWANLGRNYCGQGTQV

Ll_00958 (SEQ ID N0:#)

QVQLQESGGGLVQAGGSLRLSCARSGGTFSPYAMGVffRQAPGEERQFVAAITOSSDSPWYGDSAKGRCTIARDITKNlVYLNIFGLKPEDTAVYYCA^ , Ll_00204 (SEQ ID N0:#)

QVQLQESGGGSVQPGGSLRLSCTASMJIFNFVSMGWYRQAPGKERDLVARI . TRGGRTEYGEFV GRFTISRDNAWRTVTLQMNSLQAEDTAVYYCHIDSTMIAGIPEGYWGQGTQVTVSS

Ll_00249 (SEQ ID N0:#)

QVQLQESGGGSVQAGGFLRFSCAASGSINSITTVN YRRTSGKQLDLVARIYKSG . NTHYADSVKGRFTISRDSAENTVHLQMNSLKPEDTAVYYCKADVASSLSSYFNGRYWGQGTQVTVSS... Ll_00086 (SEQ ID N0:#)

QVQLQESGGGLVQPGGSLRLSCAASGSIFSASAMTWYRQAPGKSREYVARIFFSGG.TNYADSVKGRFTISRDNA STMYLQM NLKREDTAVYYCNLLSY GQGTQVTVSS

Ll_00350 (SEQ ID N0:#)

QVQLQESGGGLVQTGDSLRLSCTQSRRIFGSVGRGWFRQAPGKEREFVAAIT RDGNTDYADFTEGRFTISRDSAKHTIYLQMNSLKPED . Ll_00231 (SEQ ID N0:#)

QVQLQESGGGLVQPGEFLKLSCTNSGSISSIDAMGWYRQAPGKQRELVATITGAGLRT .YLSSVKGRFTISRDNAKNSMYLQMNSLKPEDTAVYFCGAGIDCSAYGCYDQKTCFYSGQGTQVTVSS Ca„00361 (SEQ ID N0;#)

QVQLVESGGGSVQTGGSLKLSCVIΞΕYT... HCASWFRQAPGADREGLANIIRDDGSTYYGTSAKGRFTIAQDNATNTWYLQMDALKPEDTATYFCAYGRKSLPGARCQIAAWGQGTQVTVSS ...

Ll_00248 (SEQ ID N0:#)

QVQLQESGGGLVQAGGFLRFSC-AASGSIGGINDMGWYRQAPGKQRELVAAITSGG. ITKYAS VKGRFWSRDSAK MYLQKDSI^PEDTAVYYCiraPKNAAKNQWGQG QV VA

Ll_00967 (SEQ ID N0:#)

...LQESGGGLVQPGGSLRLSCVASGPITSPNTMGWFRQAPGKQRELVATITYGGGTT . YADSVKGRFTISRDNAKKTMSLQKNSLKPEDTAVYYCNLW QWESESYWGQGTQVT

Ca_00086 (SEQ ID N0;#)

QVQLQESGGGSCRLEGLLKLSCTASMYIYCGYsmTWYRQAPGKEREFVAGTDLDGTTT . YADSVKGRFTISQDNAKN WLQMNSLKPEDTAMYYCKRCWYWGQGTQVTVSS

Ll_00380 (SEQ ID N0:#)

QVQLQQSGGGLVQPGGSLRLSCAASGRIFRIDVMSWYRQTPGKERELVASITSGG . SANYADFAKGRFSITIDTAKHTTIDLQ NSLKPEDSAVYYCNAKGPYARGSGPWGQGTQV VSS

Ll_00323 (SEQ ID N0:#)

QVQLQESGGGLVQLGGSLTLSCATSEDLFSTRNMAWYRQAPGKQREAVA ITTSG. TTNFANSVKGRFTISRDNAKNTVYLQmSLRPEDTAVYYCNLVHYGI YWGKR LV VSS

Ll_00861 (SEQ ID 0:#)

...LQESGGGLVQAGGSLRLSCTASGLTFGTHWGWFRQAPGNEREFVAAISYOTGSLLYADS

Ll_00524 (SEQ ID N0:#)

... LQESGGGLVQAGGSLRLSCTASGLTFGTHWGWFRQAPGNEREFVAAISYNVGSLLYADSVSGRFTISRDN KNTVYLQMVSLRPEDTAVYYCG^RAERVADANFVF,YWGQG QV

Ll_00287 (SEQ ID NO:t)

QVQLQESGGGLVQPGGFLRFSCTASGFTLDHGAIG FRQAPGQERETVSCTTPSDGAT^PDSV GRFTISREYA N VHLQMNSLKPE

Ll_00192 (SEQ ID N0:#)

QVQLQESGGGL QTGGFLRLSCTASGRTSDTLVMGWFRQVPGKEREFVAAMSWNGGRTDYADSVKGRFTISRDNDRGTLYFTINNLKPEDTAVYYCHALNRFTSALSYWGQGTQVTVSS

Ll_00918 (SEQ ID N0:#)

... LQESGGGSVQPGGSLRLSCAASGFAFSINAD YRQVPGKEREWARISSGG . ST YADAVKGRF ISRDNAKQLVFLQMNLKPEDTAVYYCNSAYKHNRDS GQGTQV

Ll_00316 (SEQ ID N0:#)

QVQLQESGGGLVQPGGSLRLSCAASGSDFSATAM WYRQPPGKSREYVARIFLSGG.TNYADSVKGRFTISRDNAKN FYLQMN LKREDTAVYYCNLASY GQGTQV VSS

Ll_00281 (SEQ ID N0:#)

QVQLQEFGGGLVQAGGFLTFSCVASGIRFSDSAMSWYRQAPGKERELVATVTLTG . SP^EDFVKGRFTISRDKAKKTVYLQM SLQPEDTAVYYC ARRPPPAYEYEY GQGTQVTVSS

Ca_00138 (SEQ ID N0:#)

QVKLEESGGGLVQPGGSLTLSCAASGFDFSSVGMTWVRQAPGKGLERVSTISALGGHTWYSDSVKGQFTISRDNAKNTLYLQLNSLETKDTA YYCSKTGLTVRTSPKAPRGQGTQVTVSS

At_QQQ63 (SEQ ID N0:#)

GGSVQAGESLRLSCVASGNIFGLNAVGWYRQAPGSQRELVARINSDS . NTIYADSVKGRF ISRMNAKDnVYLQ^SLKPEDTAWYCNWQRGRYGYEGDRNYWGQGTQV VSS

Ll_00469 (SEQ ID N0:#)

GGSVQAGESLRLSCVASG IFGL AVGWYRQAPGSQRELVARINSDS . NTIYAD-JVKGRFTISRlWAK TVYLQM SLKPEDTAVYYCliVVQRGRYGYEGDRNYWGQGTQVTVSS

Ll_00738 (SEQ ID NO:t)

DVQLQASGGGSVRAGGSLRLSCV SGSISSFDAMAWSRQAPGRQRDWAIITSGG .ATNYADSVKGRFTISRDKGNOTLYLQMNGLKPEDTAIYYCAALVASTVTSSVSWGQGTQVTVSS . ,

Ll_00781 (SEQ ID N0:#)

... LQESGGGLVQAGGSLRLSCVASGLAFENYFVS FRQPPGDEREFVAAITYSSSSTNYADSVKGRF ISRDKAVNTAYLQMD LAPGDTAVYFCAACSDYYCSGVGAVYWGQGTQVT

Ll_00024 (SEQ ID N0:#)

QVQLQQSGGGLVQPGESLRLSCSTSGFTFADYAMMWVRQAPGKGLEWVSSIDYSAEDTYYAESVKGRFT

Ll_00190 (SEQ ID N0:#)

QVQLQQSGGGLVQAGGSLRLSCVAWGNIVSTNVLRWYRQAPGKQREFVAAITSGG. LTGYSDSVNGRF ISGNNANNTAYLQMNSLKPEDTAWYCNANGRGEYWGQGTQLTVSS

Ll_00216 (SEQ ID NOi#)

QVQLQESGGGL QPGGSLRLSCAASGRILSIVKMGWYRQAPGKQRELVARL . TNGGTPQYEDSVTGRFTISRVWAGNTVFLQMNSLKPEDTAWYCKADVWVEFQRDDLWGQGTQVTVSS

Ll_00188 (SEQ ID N0;#)

QVQLQESGGGLVQPGGFLRLScaASEG FRINIMGWYRQPAGKQCELVATVATGG . HPNYADSVKGRFTISRD VKSTAYLQ NSLKPEDTAVYYCNVARLPSTGSRDYWGQGTQVTVTS

LI_01030 (SEQ ID N0:#)

... LQESGGGLVQAGDALTLSCMSGRSVDlTCAMGVffRQAPGKELEFVAAIswSGSRTEYADS™

Ll_00357 (SEQ ID N0:#)

QVQLQQSGGGLVQAGGSLRVSCAASTRAFHGSSMAWFRQAPGKEREFVARITRSGSITSYTDSVKGRFTISR TADNTITLLM NLISEDTAVYYCAAGGWG LASEYAYWGQGTQVTVSS

LI_00598 (SEQ ID N0:#)

QVQLVESGGGLVQAGGSLRLSCAASGSVSIINVMGWYRQLPGKQRELVAlJyrmAG . ^^

Ca_00419 (SEQ ID N0:#)

EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYYMSWARQAPGKGLEWVSGIYSDG . STYYGDSVKGRFTISRDNAKNMLYLQ NSLKPEDTAMYYC

Ll_00488 (SEQ ID N0:#)

... LQESGGGLVQAGGSLRLSCAASGSTFRINA^WYRQAPGKQR^

Ll_00825 (SEQ ID N0:#]

...LQESGGGLVQAGGSLRLSCAASGSTFRINAMAWYRQAPGKQRELVARISFDA. RTIYADSVQGRFTISRDNAKNTVYLQMNNLKPEDTAVYYCNQGLVWGQGTQV

Ll_00253 (SEQ ID N0:#)

QVQLQEFGGGLVQAGGFLRFSCAASGSIFRINAMGWYRQASGKERELVAl^SRD.SPNYADSVKGR^

Ll_00855 (SEQ ID N0:#)

...LQESGGGLVQAGGSLRLSCAASGDISSIVAMGWFRQAPGKERDIVARIIVffADSSYYADSVK^

Ll_00518 (SEQ ID N0:#}

...LQESGGGLVQAGGSLRLSCAASGDISSIVAMGWFRQAPGKERDIVARIIWFADSSYYADSVKGRFTISm^

Ca_00285 (SEQ ID N0:#)

EVQLVESGGGSVQAGGSLRLSCVASGPTICITDISWHRQAPGKEREFLSSIVASG. STRYAESV GRFWSRDSATNTVYLI^SLTPDDSAKYTCQSTRVGGFCYDGTWGQGTQVTVSS ... Ll_00243 (SEQ ID N0:#)

QVQLQQFGGGLVQPGGFLKFSCAASSSSISFDAMG YRPAPGKQRDLVATISSDG. TTDYSDSVKGRFTISREYAKNTVHLQMNSLKPEDTAVYYCALHLPTSTTDTGYSFWGEGTQVTVSS Ll_00303 (SEQ ID N0:#!

QVQLQESGGGLVQAGGFLTFSCAVFESILRINTKAWYRQAPGKQRELVATITSGG . STNYADSVKGRFTISRGDANTVYLRMNSLKPEDTAVYYCKVAKMIG RDYWGQGTQVTVSS

Ll_00844 (SEQ ID N0:#)

... LQESGGGLVQAGESLRLSCAASGPISGINRMNWYRQAPGKERELVASI . ]TORRINYAYSVRGRFTISRDNA NAWLQMNSIJKSEDTARYYOTALISSVSR GPFYDY GQGTQVT ... Ll_00507 (SEQ ID N0:#)

... LQESGGGLVQAGESLRLSCAASGPISGINRMHWYRQAPGKERELVASI .TNDRRINYAYSVRGRFTISRDNAKNAWLQM SLKSEDTARYYCYALISSVSRNGPFYDYWGQGTQVT ... Ll_00140 (SEQ ID N0:#)

QVQLQDFGGGLVQAGGFLRFSCAVSGTIASV AVN YRQAPGKQRELVAFITSGG . STNY DSVKGRFTISRGSANTAYLEMNSLKPEDTAVYYCNARIGLWYWGQGTQVTVSS

Ll_00790 (SEQ ID N0:#)

... LQESGGGLVQAGNSLRLSCAASGRTSDIYGMEVfflRQVPGKEREFVVILSWSDGATNYADS

Ll_00289 (SEQ ID N0:#)

QVQLQEFGGGLVQPGGFLKFSCAAFSSSISFDAMGWYRPAPGKQRELVATISSDG . TTDYSDSVKGRFTISREYPK TVHLCJ(NSLKPEDTAVYYCNALHLPTSTTDTGYSFWGEGTQVTVSS Ll_00552 (SEQ ID N0:#)

QVQLQDSGGGLVQTGGSLRLSCIVSRDIFNSY . VA FRQAPGKQRELVAGIGRDG . STTYADSVKDRFTMSRDNN DTVFLQMNSLKPEDTAVYYCYVrM3YRL GQGTQV VSS

Ll_00433 (SEQ ID N0:#)

... LQESGGGLVQPGGSLRLSCAASGSTFTNYAMG YREAPGKQRELVARISKVGD . TDYAGSVKGRFTISRDKSGNTVFLQMNSLKPEDTAVYYCNTFPAIVGPFTS GQGTQVT

Ll_^00810 (SEQ ID N0:#)

... LQESGGGLVQPGGSLRLSCAASGSTFTNYAMGWYREAPGKQRELVARISKVGD . TDYAGSVKGRF ISRDKSG VFLQMNSLKPEDTAVYYC TFPAIVGPFTS GQGTQVT

Ca_00013 (SEQ ID N0:#)

GGGSVQAGGSLRLSCTASGDSFSRFAMSWFRQAPGKECELVSSIQSNGRTTE . ADSVQGRFTISRDNSRNTVYLQMNSLKPEDTAVYYCGAVSLMDRISQHGCRGQGTQVTVSL

At_00034 (SEQ ID N0;#)

GGGSVQAGGSLRLSCTASGDSFSRFAMSWFRQAPGKECELVSSIQSNGRTTE . ADSVQGRFTISRDNSR TVYLQMNSLKPEDTAVYYCGAVSLMDRISQHGCRGQGTQVTVSL

Ll_00072 (SEQ ID N0:#)

QVQLQQSGGGLVQAGGSLRLTCAATGPIYAVNRMGWYRQAPGKQRELVATI .TNGGTIKYGDSVRGPJTISREKLN TVYLQMDNLIPEDTAIYYCTAESLRVAVDDYWGQGTQVTVSS .. Ll_01028 (SEQ ID N0:#)

... LQESGGSLVQPGGSLRLSCVASGSSVSINHMGWYRQAPGKERELVAAITSDGESA. YEDSVKGRFAISRDNLKNMVYLQMNTLKAEDTAMYYCNYLRATSAGWELRFGS GQGTQVT . Ll_00996 (SEQ ID N0:#)

... LQESGGGLVQPGGSLRLSCTASGPIFSPNTMAWFRQAPGKQRELVATITYGGGTT■ YVDSVKGRFTISRDNTKKMMSLQNNSLKPEDTAVYYCALWSQWQSETYWGQGTQVT

Ll_01063 (SEQ ID N0:#)

... LQESGGGLVQAGGSLRLSCAASASTFVINPMRWYRQAPGKQRELVAGITFHG .ATNYADSVKGRFT^ . , ..

Ll_00835 (SEQ ID N0:#)

... LQESGGGLVQAGGSLRLSCVASGRTFSLYTMGWYRQAPGKQRELVASISSQG.RTNYADSVKGRFTMSR ..VNILYLQKDSLKPEDAAVYYCHAKLFVGADS GKGTLVT

Ll_00498 (SEQ ID N0:#)

... LQESGGGLVQAGGSLRLSCVASGRTFSLYTMGWYRQAPGKQRELVASISSQG .RTNYADSVKGRFTMSR ..VKNILYLQMDELKPEDAAVYYCHAKLFVGADS GKGTLVT

Ll_00502 (SEQ ID N0:#)

... LQESGGGLVQPGGSLKLSCAASGFTFDDYAIGWFRQAPGKEREGVICI NDGD. THYADFWGRFTISRmA^TWLQTINELKPDDTAWHCGKLiRSQIWGAPIEEYNHWGRGTQVT

Ll_00839 (SEQ ID N0:#)

... LQESGGGLVQPGGSLKLSCAASGFTFDDYAIGWRQAPGKEREGVICINNIX3D. THYADFVKGRFTISRDNAR TW

Ll_0O09O (SEQ ID N0:#)

QVQLQESGGGLVQAGGSLTLSCASSRSISSIDYMMHYRQAPGKERDMVARIDSDG . STOTADSVKGPJTISRKlARDTLYLQMNNLSPEDTAVYSCTAWASGLKILQY GOGTO TVSS

Ll_00923 (SEQ ID NO:#)

... LQESGGGLVQTGGSLRLSCAASGFAFSINAMDWYRQVPGKEREVVARISSGG . STNYADAWGRFTISPJDNAQLWLQt-NHLKPEDTAVYYCNSAYKHNRDSWGQGTQVT

L1_0014B (SEQ ID NO:#)

QVQLQQSGGGWEPGESLRLVCTSSGSIFSINAMGWYRQAPVKQRELVADISS .GDΞ^WAEΞVKGRF IΞRDFRKRTWLQ!NNLKPEDTAIYYCARGITDYYRE DEL· GQGTQV VSS

Ca_0024O (SEQ ID NO:#)

AVQLVESGGGLVQPGGSLRLSCAASGFRFSGVITCSWVKQAPGKGLDWVSDINGDGDSIHWGSVKGRFTI^

Ll_00439 (SEQ ID NO:#)

... LQESGGGLGQAGGSLRLSCAASGTVSNIKFMAWYR.QAPGKQREFLATIISDG .NTRYADSV GRFTINRDNAENTAALQK SLKPEDTAVYYCNARYGRFLIDY GQGTQVT

Ll_00816 (SEQ ID NO:#)

...LQESGGGLGQAGGSLRLSCAASGTVSNIKFMAWYRQAPGKQREFLATIISDG .NTRYADSVKGRFTINRDNAE TAALQKNSLKPEDTAVYYCMARYGRFLIDY GQGTQVT

LI_^00193 (SEQ ID NO:#)

QVQLQESGGGLVQPGGFLRFSCAASGTTASINDMGWYRQAPGKQRELVAYIDIDG.HANYTNSAIGRFTI^

LI_00208 (SEQ ID NO:#)

QVQLQEFGGGI-AQPGGSLRLSCVASGFTLDYYAITWFRQVPGKEREGISCISPSNGNTIYADS™

At_00045 (SEQ ID NO:#>

GGGSVQVGGSLKLSCKISGGTPDrkSLAVERQAPEKEREGIAVLSTKLX.KTFYADSVKGRFTIFLDND ....NYWLYAIWGQGTHVTVSS

Ca_00006 (SEQ ID NO:#)

GGGSVQVGGSLKLSCKISGGTPDrkSI^ FRQAPEKEREGIAVLSTKDGKTFYADSVKGPJTIFLDNDKTTFSLQLDRI^PEDTADYYCAAQI^GG YLDP ....NYWLYAIWGQGTHVTVSS

Ca_00328 (SEQ ID NO:#)

EVQLVESGGGSVHVGGSLRLSCAASGSTSRLFCl^VreRHVPGKERERVAAmy TTIKTYYHDSVKGRFALSRDNAGPTWYLQMDDLKL

Ll_00145 (SEQ ID NO;#)

QVQLQESGGGLVQAGGSLRLSCTASLDIVNINIMGWYRQAPGKQRDLVAITiPKRGDS .NYADSVKGRFTISRGSAKStVYLQMDNLKPEDTAVYYCWQYYIGT

L1_0023Q (SEQ ID NO:#)

QVQLQEFGGGLVQPGGFLHFFCVASGNIPRTYFMSWYRRPPGKERELVASMPLAGNFI . YADSVKGRFTISRDDANTWLQMNDLKLEDTAVYTCAKDATCGGWNHPTYTGFDYWGKGTLVNVST

Ll_00493 (SEQ ID NO;#)

... LQESGGGLVQAGGSLRLACAASVSISTIYVMGWYRQAPGKQRELWSVQ .NGIYTNYADSVKGRFTISRDDANTLYLQMSSLKPEDTAVYYCAAIYTPGRGEFDYWGQGTQV

LI_Q0830 (SEQ ID NO:#)

... LQESGGGLVQAGGSLRLACAASVSISTIYVMGWYRQAPGKQRELWSVQ .NGIYTNYADSVKGRFTISRDDANTLYLQMSSLKPEDTAVYYCAAIYTPGRGEFDYWGQG QV

Ll_00829 (SEQ ID NO:#)

... LQQSGGGLVQPGGSLRLSCaaSGNAFRINALGWYRQAPGKQRELVALIASNG.NTHYVDSVKGRFTISKDNAKNTVYLQMSSLKPEDTAVYTCAVDNNPIVYRPNDYWGQGTQV

Ll_00492 (SEQ ID NO:#)

... LQQSGGGLVQPGGSLRLSCaaSGNAFRINALGWYRQAPGKQRELVALIASNG.NTHYVDSVKGRFTISKDNAKNTVYLQMSSLKPEDTAWTCAVDNNPIWRPNDYWGQGTQVT

LI_00089 (SEQ ID NO:#)

QVQLQESGGGLVQPGGSLRLSCAASGSISSATAMTWFRQAPGKSREYVARIFFSGG.TNYADSVKGRFTIARDAAK^^

Ll_00286 (SEQ ID NO:#)

QVQLQQSGGGLVQPGGFLRFSCAASGFRLDNYGIGWFRQAPGqeHEAVACISSRKGVTYYSDPVKGRFTISRDATNNAVNLQMNSLKPEDTAVYYCTTEPWVCPSAASDYADTYREVRGQGTLVTVSS ... At_00075 {SEQ ID NO:#)

GGSVQAGGSLRLSCVHSGTTECYYDMSWYCQAPGKEREFVSGIDSDEGSR.YADSAGRFTISQDNAKNTVLLQM LKPEDTAMYYCPiWKWCPRRLTCLTYWGQGTRVTVSS

Ll_00456 (SEQ ID NO:#)

GGSVQAGGSLRLSCVHSGTTECYYDMSWYCQAPGKEF-EFVSGIDSDEGSR.YADSAKGRFTISQDNAK^

L1_QQ278 (SEQ ID NOi#)

QVQLQQFGGGLVQAGGFLRFSCAASGFTFDDYALGVffRQAPGKERAGISCVSTKDGTAYYADSVKDRFTIST^

Ll_00438 (SEQ ID NO:#)

... LQQSGGGLVQAGDSLRLSCTYSGVGFSV NLGWFRQAPGKEREFVASMMWSGGS . DYVDSVKGRFTISRDNAKNTATLQMHSLKPEDTAVYFCAAQAAGLSRDAHEYKYWGQGTQVT

Ll_00815 (SEQ ID NO:#)

... LQQSGGGLVQAGDSLRLSCTYSGVGFSVTNLGWFRQAPGKEREFVASMMWSGGS . DYVDSVKGRFTISRDNAKNTATLQMNSLKPEDTAVYFCAAQAAGLSRDAHEYKYWGQGTQVT

98

100

Ca_00320 (SEQ ID N0:#)

QVQLVESGGGSAQAGGSLRI-ACGASGYTDRYCTMGWYRQAPGKQRELVAOTIKNS .RPW^

Ca_00416 (SEQ ID N0:#)

EVQLVESGGG-TVQAGGSLRLSCTAPGFTSNSCGKDOTRQAAGKQREVin/SSISTDG.STSYADSVKGRFTISKDKAKDTVYLQMNSLKPEDTAMYYC

Ll_0022B (SEQ ID N0:#)

QVQLQESGGGTVQAGGSVSLSCTASGNFFELRT GWYRQTPGSQRELVADITGVG. R NYAGSAKGRFTISRDNAK TVYFQMNNLKPEDTGVYYCNADYSLYWGQGTQVTVSS

At_00049 (SEQ ID N0:#)

GGGSVQAGGSLRLSCVASGFNFETSRMAWYRQTPGWCELVSSIYSDG.KTYYVDRlffi^

Ca_00017 (SEQ ID N0:#)

GGGSVQAGGSLRLSCVASGFNFETSRHAWYRQTPC5HVCELVSSIYSDG.KTYYVDRM GRFTISRENA NTLYLQLSGLKPEDTAMY

Ll_00912 (SEQ ID N0:#)

... LQQSGGGLVQPGGSLKLSCAASG . SISRNGMD YRQVPGKEREWVATITSRG.NTNYADSVKGRFTISRDLA NMVDLQKNSL PEDTAVYYCR RGY GQGTQVT

L1_Q0511 (SEQ ID N0:#)

... LQESGGGET7PVGGSLRLSCVASGIVFSSHAMNWYRQVPGKERGLVAHITTTG. S lϊrrSNPVKGRASIS DNAKHTWLQ^_DSLKPED AW CNA LGDS VDEYWGQG QVT

L1_00S48 (SEQ ID NO: #)

... LQESGGGSVPVGGSLRLSCTASGIVFSSHAMNWYRQVFGKERGLVAHITTTG. S MYSNPVKGRASISRDNAK TVYLQKDSLKPEDTAVYYCNAVKLGDSTVDEYWGQGTQVT

Ll_01067 (SEQ ID NO: #)

...LQESGGGLVQPGGSLRLSCAASGSFGYIGGMGWYRQTPGKQREFVAWITSDL . SADYADSWG F ISRDNARHTWLE^-WL PEDΊΪVYYC AR PSSS ILDDWGQG Q T

L1_Q0605 (SEQ ID NO:#(

QXXXQESGGGLVQAGGSLRLSCAASEGTISSYVMGWFRQAPGKEREFVAAITRSGNNTYYTDSLKGRFTISRENAKNTVYLQMNSLRMSDFGS GQGT

Ll_00176 (SEQ ID NO:#)

QVQLQESGGGLVQAGGSLRLSCAASGSIFRIDAMGWARAAPGKQREFVVRVDSGG.VAKYADSVKDW

At_00081 (SEQ ID NO:#)

GGSVQAGGSLTLSCTASGH WNSYVMGWRQTPGKEREGVAVISIGFGSTFYADSVKGRFTIREQRG N

Ll_00464 (SEQ ID NO;#}

GGSVQAGGSLTLSCTASGHTTOISYVMGWFRQTPGKEREGVAVISIGFGSTFYADSVKGPJTIREQRGMNTVH^

Ll_01002 (SEQ ID NO:#)

...LQESGGGLVQPGGSLRLSCAASGSDFSATAMTWYRQPPGKSREYVARIFLSGG . TNYADSVKGRFTISRDNA-OTTFYLQtDJHLKREDTAVYYCNLASYWGQGTQVT

Ll_00895 (SEQ ID NO:#)

... LQESGGGLVQAGGSLTLSCASSRSISSIDYM WYRQAPGKERDMVARIDSDG. ST^ADSVKGRFTISRDGARDTLYLQtJ^LSPEDTAVYSCVAWASGLKILQYWGQGTQVTVSSEPKTPKPQP .. Ca_00412 (SEQ ID NO:#)

QVKLEESGGGSVQAGGSLRLSC APGFTSNRCGMD YRQAPGKEREFVSSISTDG. TTSYADSV GRFTISKDKAKDTVYLQMNSLKPEDTAMYYC

Ll_00033 (SEQ ID NO:#)

QVQLQESGGGSVQPGGSLRLSCATSGFTFDRSWMYWLRQPPGKQIEWVASVNWDASQINFVNSV^

Ll_00279 (SEQ ID NO:#)

QVQLQQSGGGLVQAGGFLRFSCAASGRISSIDTMGWYRQTPGNQREFVATI . TKTYCRDYADSVKGRV IARDSANN. IWLQMNNLKPEDTAVYYCQAFAYSQQLSGLVAYWGQGTQVTVSS

Ll_0-020 (SEQ ID NO:#)

... LQESGGGLVQPGGSLRLSCAASGSIFSP DMGWYRQAPGLLRELVARIDYSGHT .YADSVKGRF ISRDTTKKTMYLEMNSLKPEDSGVYYCGLFSK SSDYWGQGTQVT

Ll_00471 (SEQ ID NO:#)

GGWQPGVSLGLSCMSGFTFSTDWMYWVRQAPGKGLEWVSTINDAGDRT^QDSVKGRFTI^

Ll_00778 (SEQ ID NO:#)

... LQESGGGLVRPGESLRLSCWSGSDINFNVMGWYRQAPGQQRELVA I SGG. STDYADSVKGRFTISRHNAKNMVYLQMNSLKPEDTAVYYCNADIFFVNY GKGTLVT

Ca_00087 (SEQ ID NO;#)

QVQLQESGPGLVKPSQTLTLTCTVSgiTTSYYGWSWIRQPPG GLEWMGAIAYSG. STYYSPSLKSRTSFSRDTSKNQFSLQLSSVTPEDTAVYYCARDSPRLRVGSESR... GVYSMDY GKGTLVTISS Ll_00638 (SEQ ID NO:#)

QXQLVESGGGLALAGDSLRLSCAASXRTFSXYTMG FRQAPXKEREFVXAISXTGGSTYYADSMKGRFTISRDNAKNXX Q^mLKPED A YXN XF Y R TYWGQGTQVTVSS

Ll_00283 (SEQ ID NO;#)

QVQLQQSGGGSVQPG3FLRFSCTTFGTIEHI T GWYRQAPGKERELVAVTLTDG.VTKYADSVKGRFTMSRHNGKNTGYLQMESLKPE

Ll_00766 (SEQ ID NO:#)

...LQESGGGWVQPGGSLRLSCAASEMIFDFNDMGWYRQAPGKERELVASISRFG. RTNYIDSVKGRFSISNDPA W VYLQMNSLKPEDTAVYSCNTDPPLFAWGQGTQVT

103

104 Ca_00368 (SEQ ID N0:#)

QVKLEESGETSVMAGGSLRLSCSVSDLTFDNVHMGWYRKTPGSECEMVAEIKSSG. EIWYHMSVQGRFTISRVNEKRTAYLQMDNLAPSDTAMYYCSAEGVDWFKGGCSGRWSQGTRVTVSS

Ll_00940 (SEQ ID N0:#)

EVQLVESGGGLVQGWGLSELSCAASGRTIN YAMGWYRQAPGKQRELVAGITRGG..KHELCSLrgPIPPsrDNAKNTVYLQM QLKTLRtpVYYCAAKRGPIIRRIVLRYNYWGQGIQVTVSSAAASGSF At_00054 (SEQ ID N0:#)

GGLVQPGGSLRLSCAASGFAFSTYAilNWVRQTPRYGLEWVSCIDSTGDTTIEIESVKGRF

L1_^00479 (SEQ ID N0:#)

GGLVQPGGSLRLSCAASGFAFSTYA N WQ PRYGLEWVSCIDSTGDTTIEIESV GRFTISRD!OTKNTLYLQMSNLRPEDSARYYC A

Ll_00907 (SEQ ID N0:#)

... LQQSGGGLVQTGGSLRLSCAVPESTFSITSMGWYRQASGKERELVAGIISAG . TANYADSVKGRFTISIGNTKNTAYLQM SLKPGDTAVYYCRWGTYKGD

Ll_00450 (SEQ ID N0:#)

GGLVQPGRSLRVSYAASGFTFSSHYMSWVRQDPEKGLE VSEIATGG ITSYADSVKGRFTISRDNA NMLFLQMNNLKPEDTALYYCVRRGRAIAFDVWGQGTQVTVSS

At_00055 {SEQ ID N0:#)

GGLVQP03SLTLSCKVSGFTSDEYIIGWFRRVPGKEREGVSCISFJCEGTTYYGDGMRGRFTISGDOT

Ll_^00478 (SEQ ID N0:#)

GGLVQPGGSLTLSCKVSGFTSDEYIIGWFRRVPGKEREGVSCISRKEGTTYYGDGIOIGRFTISGD VASTWLRMSNLKPEDTAVYYCAAEPRPRC^

At_00080 (SEQ ID N0:#)

GGSVQAGGSLRLSCSASTIDYRPTWGWFRQAPGKEREWAGWIGAESGSYSKDVKDRFTVSQDTAKRTVYL

Ll_00465 (SEQ ID NO:#)

GGSVQAGGSLRLSCSASTIDYRPTWGWFRQAPGKEREWAGWIGAESGSYSKDVKDRFWSQDTAKRTVYLQMTSI^

Ll_00224 (SEQ ID N0:#)

QVQLQESGGGLVQiKFLRFHCVLSGGPVSTRIiMAWPRQAPGKEREFVASMTSEG.TPNYSFAATGRFTISRDmRSTMDLQMDSLQPEDTAWYCALANGPR

Ll_00803 (SEQ ID N0:#)

... LQESGGGWQPGESLTLSCVASGTFQRINHKAWYRQIPGKEREQVAIIHNDGRL .NYAASVKGRFTISRDSAENTAHLQKKSLRPEDTAVYYCNVDLDVGFITEHYWGPGTQVT

L1_D0482 (SEQ ID N0:#)

GGLVRPGGSLRLSCAASGITLDYYAIAWFRQAPGKEREQISCIRTRGED YHLPSGKDRFTISRDNYNNTVYLFJMNNLRP

At„00058 (SEQ ID N0:#)

GGLraPGGSLRLSCAASGITLDYYAIAWFRQAPGKEREQISCIRTRGEDTYHLPSGKDRFTISRDITOmWLHI^

Ll_00801 (SEQ ID N0:#)

... LQESGGGSVQPGGSLRLSCATSGFTFDRSViMYWLRQPPGKQIEWVASVNVTOASQINFWSVKD

Ca_00133 (SEQ ID N0:#)

QVKLEESGGGSVQVGGSLTLSCTVPGLTANRCGVSWYRQVPGGQRR ISSITANGITRSYSPSLKGRFTISLDNSG

At_00078 (SEQ ID N0:#)

GGSVEAGGSLRLSCTRSGASNPKYYIAWFRQAPGKERERLLAINDAGWTYGSDNLKDRAAISQDAakDTVYLEMNlJLKPEDTAIYTCAAGPRHGCNVWVFNYWGPGTQVTVSS

Ll_00461 (SEQ ID N0:#)

GGSVEAGGSLRLSCTRSGASNPKYYIAWFRQAPGKERERLLAINDAGWTYGSDNLKDRAAISQDAakDTWLE NNLKPEDTAIYTCAAGPRHGCNVWVFNYWGPGTQVTVSS

Ll_00802 (SEQ ID N0:#)

... LQESGGGLVPTGGSLTLSCEVSGNIFSLNTMRWYVQTPGNEREMVASITS. RKIPKYADSVEGRFTISRDNALNTIDLQHNSLKPEDTGVYYCNGDVHDGMQLRNYWGQGTQVT

Ll_01031 (SEQ ID N0:#)

... LQESGGGLVQAGGSLRLSCAASG SCIS..DGSTHYADSVKGRFTISSDNA NTVYLQMNSLKPEDTAVYYCAADSDDSNCQISWYDYWGQGTQVT

Ll_00776 (SEQ ID N0:#)

... LQQSGGGFVQPGGSLRLSCTASGRTSTV GMGWYRQAPGKLRERVAWMPRG. TTQYHDNVKGRFTISRDNTRTTAYLQMNDLQPDDTANYYCFADVGPTVKVGQGTQVT

Ll_00983 (SEQ ID N0:#)

... LQESGGGFVQPGGSLRLSCAASG SVIDSGGSSTNYADEWKGRFTISRDNAKN LYLQHNSLRSADTAVYYCAKARGRGFVSGPDSLDRGQGTQV

At_00057 (SEQ ID N0:#)

GGLVQTGGSLRLSCAASKTLHSISDMAWYRQPPGKRRKGVATISTDG. STDYDDSVRGRFTI

Ll_00483 (SEQ ID N0:#)

GGLVQTGGSLRLSCAASKTLHSISDMAWYRQPPGKRRKGVATISTDG. STDYDDiTVRGRFTITG NAENTISLQM^LRPEDSAVYFCQAVIVSVIRQ PG GQGAQVTVIS

Ca_00122 (SEQ ID N0:#)

EVQLVESGGGSVQAGGSLRLSCRAAGPVPRRGElgWFRQAQGENCEELAQL . LDTNGPKYG. SLlcgRFTLTRDNTESASFLQMDNLGPEDTAIYYCMAAHQAIYDGADRRDRWKRQLQDGSACARAG DYW

Claims

CLAIMS What is claimed is

1. An ASCP-body comprising a bottomside epitope binding domain comprising Bottomside Binding Region ("BBR") 1, BBR 2, BBR3, and BBR4.

2. The ASCP-body of claim 1, wherein the bottomside epitope binding domain binds to a half-life extender.

3. The ASCP-body of claim 2, wherein the half-life extender is human serum albumin.

4. A pharmaceutical composition of the ASCP-body of claim 1.

5. The ASCP-body of claim 1, wherein the ASCP-body further comprises a topside epitope binding domain comprising complementarity determining region ("CDR") 1, CDR2 and CDR3.

6. The ASCP-body of claim 5, wherein the topside epitope binding domain binds to the same antigen as the epitope bound by the bottomside epitope binding domain.

7. The ASCP-body of claim 5, wherein the topside epitope binding domain binds to a different antigen as the epitope bound by the bottomside epitope binding domain.

8. The ASCP-body of claim 5, wherein the topside epitope binding domain binds to a half-life extender.

9. The ASCP-body of claim 5, wherein the half-life extender is human serum albumin.

10. The ASCP-body of claim 5, wherein the bottomside epitope binding domain binds to a half-life extender.

1 1. The ASCP-body of claim 10, wherein the half-life extender is human serum albumin.

12. A pharmaceutical composition of the ASCP-body of claim 5.

13. A composition comprising a first ASCP-body of claim 5, operably linked to one or more additional ASCP-bodies of claim 5.

14. The composition of claim 13, wherein the topside epitope binding domain of the first ASCP-body and the topside epitope binding domain of the one or more additional ASCP-bodies bind to the same antigen.

15. The composition of claim 13, wherein the topside epitope binding domain of the first ASCP-body and the topside epitope binding domain of the one or more additional ASCP-bodies bind to different antigens.

16. The composition of claim 13, wherein at least one of the topside or bottom side epitope binding domains binds to a half-life extender.

17. The composition of claim 13, wherein the half-life extender is human serum albumin.

18. A pharmaceutical composition of the ASCP-body of claim 13.

19. A composition comprising one or more first ASCP-bodies of claim 1, operably linked to one or more second ASCP-bodies, wherein the one or more second ASCP-bodies comprise a topside epitope binding domain comprising CDR (complementarity determining region) 1 , CDR2 and CDR3.

20. The composition of claim 19, wherein the bottomside epitope binding domain of the one or more first ASCP-bodies binds to the same antigen as the topside epitope binding domain of the one or more second ASCP-bodies.

21. The composition of claim 19, wherein the bottomside epitope binding domain of the one or more first ASCP-bodies binds to a different antigen than the topside epitope binding domain of the one or more second ASCP-bodies.

22. The composition of claim 1 , wherein at least one of the topside or bottom side epitope binding domains binds to a half-life extender.

23. The composition of claim 1 , wherein the half-life extender is human serum albumin.

24. A pharmaceutical composition of any of the ASCP-body of claim 1 .

25. A composition comprising a first ASCP-body comprising a bottomside epitope binding domain comprising BBR (Bottomside Binding Region) 1, BBR 2, BBR3, and BBR4, operably linked to one or more additional ASCP-bodies comprising a bottomside epitope binding domain comprising BBR (Bottomside Binding Region) 1, BBR 2, BBR3, and BBR4.

26. The composition of claim 25, wherein the bottomside epitope binding domain of the first ASCP- body and the bottomside epitope binding domains of the one or more additional ASCP-bodies bind to the same antigen.

27. The composition of claim 25, wherein the bottomside epitope binding domain of the first ASCP- body and the bottomside epitope binding domains of the one or more additional ASCP-bodies bind to different antigens.

28. The composition of claim 25, wherein at least one bottomside epitope binding domain binds to a half-life extender.

29. The composition of claim 25, wherein the half-life extender is human serum albumin.

30. A pharmaceutical composition of the ASCP-body of claim 25.

31. A composition comprising a ASCP-body of claim 1 , operably linked to a non-ASCP-body entity.

32. The composition of claim 31 , wherein the non-ASCP-body entity is one or more proteins, peptides, carbohydrates and/or lipids.

33. A library of ASCP-bodies comprising a bottomside epitope binding domain comprising BBR (Bottomside Binding Region) 1, BBR 2, BBR3, and BBR4.

34. A method of creating the library of claim 33, the method comprising the steps of providing a V_HH domain directed to a desired antigen,:

(a) by a method comprising the steps of:

(i) immunizing a mammal belonging to the Camelidae with the desired antigen or a fragment thereof, so as to raise an immune response and/or antibodies (particularly heavy chain antibodies) against the antigen;

(ii) obtaining a biological sample from the immunized mammal, wherein the sample comprises heavy chain antibody sequences and/or V_HH sequences directed against the antigen; and

(iii) obtaining the BBR sequences from the heavy chain antibody sequences and/or V_HH

sequences; or

(b) by a method comprising the steps of

(i) screening a library comprising heavy chain antibody sequences and or VHH sequences for sequences directed to the antigen;

(ii) obtaining the heavy chain and/or V_HH sequences from the library; and

(iii) obtaining the BBR sequences from the heavy chain and/or V_HH sequences.

35. The method of claim 34, further comprising the step of (b) mutagenizing the VHH domain.

35. A composition comprising the ASCP-body of claim 1 , further comprising a CH 1 , CH2, and/or CH3.

36. The ASCP-body or ASCP-body composition of any of the above claims, wherein the composition is humanized.