CN108129567B - A kind of Humanized anti-human CD146 monoclonal antibody having efficient neutralization activity - Google Patents
A kind of Humanized anti-human CD146 monoclonal antibody having efficient neutralization activity Download PDFInfo
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Abstract
The invention discloses a kind of Humanized anti-human CD146 monoclonal antibodies for having efficient neutralization activity, and the Humanized anti-human CD146 monoclonal antibody for having efficient neutralization activity includes light chain variable region and heavy chain variable region.The present invention is under the premise of keeping or enhancing antibody molecule is active in conjunction with people's CD146 acceptor molecule, prepare the high monoclonal antibody of humanization degree, the caused anti-mouse of people is immunoreacted after entering human body as therapeutic antibodies to avoid the antibody, has reached good technical effect.
Description
Technical field:
The present invention relates to biomedicine fields, and in particular to a kind of Humanized anti-human CD146 for having efficient neutralization activity
Monoclonal antibody.
Background technique:
CD146 is a kind of adhesion molecule with transmembrane structure, the I type memebrane protein being made of 646 amino acid residues,
Belong to Mitochondrion IgG superfamily member.Research shows that CD146 is being total to for vascular endothelial growth factor receptor VEGFR-2
Receptor is adjustable the activation of the VEGFR-2 of VEGF induction and the transmitting of downstream signal, and then promotes Tumor Angiongesis
(MicroRNA 329 Suppresses Angiogenesis by Targeting CD146.Molecular
and Cellular Biology,2013;33(18):3689-99.).In addition, CD146 molecule is that inflammatory bowel disease is promoted to be converted into
An important factor for colorectal carcinoma, pro-inflammatory cytokine TNF α and IL1 β can activate the transcription of CD146, make it in the blood vessels
High expression on chrotoplast film, and then lead to the continuing advances of chronic inflammation by promoting inflammatory cell to invade profit and angiogenesis, it drills
Become colorectal cancer.By a series of relevant tumor model of ulcerative colitis, researcher has found specific knockdown blood vessel
The CD146 of endothelial cell, makes the frank mitigation of inflammatory bowel disease mouse, inflammatory cell invade profit and new vessels are obvious
It reduces;In the blood vessels in the tumor model of chrotoplast specific knockdown CD146, the quantity and volume of tumour can be also decreased obviously
Targeting endothelial CD146 attenuates neuroinflammation by limiting
lymphocyte extravasation to the CNS.ScientificReports,2013;3:1687..CD146 molecule
Being overexpressed in the breast cancer cell of epithelial character makes the lower tumour cell of script grade malignancy obtain high cell invasion
Transfer ability, and show the feature of breast carcinoma stem cell.Research shows that CD146 by the mechanism of EMT promote breast cancer into
Exhibition, therefore CD146 may be as new target drone (CD146, an an epithelial-mesenchyaml for the treatment of breast cancer
transition inducer,isassociated with triple-negative breastcancer.Proceedings
of the National Academy of Sciences of the United States of America,2012;109
(4)1127-32.)。
It is the antibody of target spot because of its structure feature difference using CD146, causes it from the affinity of CD146 molecule and different.
P1H12 can in conjunction with most of different types of vascular endothelial tissues (Identification of the S-Endo1
endothelial-associated antigen.Biochem Biophys Res Commun.1996;218(1):210-
6.);Some anti-CD146 antibody only combine the blood vessel that active growth state is in tumor tissues, do not combine in normal tissue
Blood vessel.Research shows that these antibody identify that different epitopes, P1H12 identify the linear list for being located at N-terminal structural domain
Position, and some anti-CD146 antibody identify the space conformation epitope based on disulfide bond, therefore cause to different endothelial tissue knots
Close active difference.CN102936283A also discloses a kind of source of mouse CD146 antibody, being capable of the above-mentioned people CD146 of specific recognition
Epitope.
Above-mentioned several antibody are mouse monoclonal antibodies, low with human complement component binding ability, to tumour cell
Killing ability is weaker;Weak with the immunocytes surface Fc receptors affinity such as NK, the ADCC effect of mediation is weaker;And source of mouse antibody
Half-life short in people's blood circulation has immunogenicity, thus host is also easy to produce antiantibody and causes rejection etc. many not
Foot.
Currently, lacking a kind of Humanized anti-human CD146 monoclonal antibody for having efficient neutralization activity.
Summary of the invention:
In view of this, proposing the present invention.
The first aspect of the present invention provides a kind of light chain variable region of Humanized anti-human CD146 monoclonal antibody, described
Chain variable region amino acid sequence is any of SEQIDNO:12-17.
Another aspect of the present invention provides a kind of heavy chain variable region of Humanized anti-human CD146 monoclonal antibody, described
Heavy chain variable amino acid sequence is any of SEQIDNO:1-11.
It is anti-to provide a kind of Humanized anti-human CD146 monoclonal for having efficient neutralization activity for another aspect of the present invention
Body, the monoclonal antibody include at least above-mentioned light chain variable region, one kind of heavy chain variable region.
Further, above-mentioned Humanized anti-human CD146 monoclonal antibody further includes heavy chain constant region and constant region of light chain, institute
The amino acid sequence for stating heavy chain constant region is amino acid sequence shown in SEQIDNO:18;The amino acid sequence of the constant region of light chain
Column are amino acid sequences shown in SEQIDNO:19.
Another aspect of the present invention provides a kind of above-mentioned Humanized anti-human CD146 monoclonal antibody in preparation for treating
Application in malignant tumour or Autoimmune diseases or new vessels class disease medicament.
Another aspect of the present invention, provides a kind of for treating the pharmaceutical composition of malignant tumour, and it includes above-mentioned source of people
Change anti-human CD146 monoclonal antibody and pharmaceutical carrier.
Another aspect of the present invention provides a kind of above-mentioned Humanized anti-human CD146 monoclonal antibody in preparation for detecting
Or the kit or the application in diagnosticum of diagnosing malignant tumor or Autoimmune diseases or new vessels class disease.
Above-mentioned malignant tumour is selected from irradiation, glioma, nasopharyngeal carcinoma, cancer of pancreas, lung cancer, the cancer of the esophagus, mammary gland
Cancer, gastric cancer, colorectal cancer, liver cancer, prostate cancer, malignant tumor of ovary, cervical carcinoma, carcinoma of endometrium, malignant mela noma, skin
Cancer, lymthoma, leukaemia or thyroid cancer.
Above system autoimmunity disease is selected from systemic loupus erythematosus, rheumatoid arthritis, systemic vasculitis, hard
Skin disease, pemphigus, dermatomyositis, mixed connective tissue disease, autoimmune hemolytic anemia, Autoimmune Thyroid Diseases or ulcer
Property colitis.
Above-mentioned new vessels class disease is selected from age-related macular, multiple sclerosis, macular edema, choroidal neovascularization
Disease, neovascular glaucoma, retinal vein obstruction or retina neovascular diseases.
Another aspect of the present invention provides the Humanized anti-human CD146 for having efficient neutralization activity described in a kind of coding
The nucleic acid sequence of monoclonal antibody light chain variable region, the nucleic acid sequence are any of SEQIDNO:31-36.
Another aspect of the present invention provides the Humanized anti-human CD146 for having efficient neutralization activity described in a kind of coding
The nucleic acid sequence of monoclonal antibody heavy variable region, the nucleic acid sequence are any of SEQIDNO:20-30.
Another aspect of the present invention provides the Humanized anti-human CD146 for having efficient neutralization activity described in a kind of coding
The nucleic acid sequence of monoclonal antibody constant region of light chain, the nucleic acid sequence are SEQIDNO:38.
Another aspect of the present invention provides the Humanized anti-human CD146 for having efficient neutralization activity described in a kind of coding
The nucleic acid sequence of monoclonal antibody heavy constant region, the nucleic acid sequence are SEQIDNO:37.
Another aspect of the present invention provides the Humanized anti-human CD146 for having efficient neutralization activity described in a kind of coding
The nucleic acid sequence of monoclonal antibody, the nucleic acid sequence are any of the above-described kind of light chain variable region, heavy chain variable region, chain constant
The combination in area, heavy chain constant region nucleic acid sequence.
Further, the present invention provides a kind of expression vector, and the expression vector includes encoding SEQIDNO:20-36 at least
A kind of nucleic acid sequence.Preferably, the expression vector includes at least one of SEQIDNO:20-30 and SEQIDNO:31-36
At least one of.
Further, the present invention provides a kind of protokaryon or eukaryotic host cell including above-mentioned expression vector.
The invention has the benefit that Humanized anti-human CD146 monoclonal antibody of the invention has efficient neutralization activity,
Under the premise of keeping or enhancing antibody molecule is active in conjunction with people's CD146 acceptor molecule, it is high as far as possible to prepare humanization degree
Monoclonal antibody, enter the caused anti-mouse immune response of people after human body as therapeutic antibodies to avoid the antibody;The present invention
Using experiment with calculating the method combined in monoclonal antibody surface progress rite-directed mutagenesis, the framework of human IgG is selected, not
Change in original antibody CDR structure basis, rite-directed mutagenesis is carried out to the area framework of IgG;Antibody structure packet prepared by the present invention
The heavy chain variable region of the IGHV3-33 containing people and the area frame FR of light chain variable region, do not influence antibody target tropism and it is functional before
It puts, point mutation is carried out to framework region, to improve the combination activity of antibody and people's CD146 receptor.
Detailed description of the invention:
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below
There is attached drawing needed in technical description to be briefly described, it should be apparent that, the accompanying drawings in the following description is only this
Some embodiments of invention for those of ordinary skill in the art without creative efforts, can be with
It obtains other drawings based on these drawings.
Fig. 1 is ELISA result of the anti-CD146 source of mouse antibody in conjunction with CD146.
Specific embodiment
Example embodiments are described in detail here, and the example is illustrated in the accompanying drawings.Following description is related to
When attached drawing, unless otherwise indicated, the same numbers in different drawings indicate the same or similar elements.Following exemplary embodiment
Described in embodiment do not represent all embodiments consistented with the present invention.On the contrary, they be only with it is such as appended
The example of device and method being described in detail in claims, some aspects of the invention are consistent.
It is only to be not intended to limit the invention merely for for the purpose of describing particular embodiments in terminology used in the present invention.
It is also intended in the present invention and the "an" of singular used in the attached claims, " described " and "the" including majority
Form, unless the context clearly indicates other meaning.It is also understood that term "and/or" used herein refers to and wraps
It may be combined containing one or more associated any or all of project listed.
It will be appreciated that though various information, but this may be described using term first, second, third, etc. in the present invention
A little information should not necessarily be limited by these terms.These terms are only used to for same type of information being distinguished from each other out.For example, not departing from
In the case where the scope of the invention, the first information can also be referred to as the second information, and similarly, the second information can also be referred to as
One information.Depending on context, word as used in this " if " can be construed to " ... when " or " when ...
When " or " in response to determination ".
The present invention will be described in detail by way of examples below.
The Humanized strategies of the anti-human CD146 human mouse chimeric antibody of embodiment 1
Anti-human CD146 human mouse chimeric antibody is carried out using best-fit CDR constructing technology humanization modified.Source of mouse
CDR, which is transplanted to, to be suitble in frame human-acceptable with chimeric antibody framework region with high sequence homology.In people
The FR frame of most matched portable CDR is searched in Germline sequences database.It is tied using the primary structural sequence and 3D of antibody
Structure data maintain the correct conformation of source of mouse CDR so that critical framework needed for keeping binding affinity and binding specificity is residual to determine
Base.Back mutation is carried out in different humanized molecule's frames to keep the affinity of antibody, finally obtains height source of people
The monoclonal antibody of change.
The database of the area the V gene of people, people IGHV3-33 and anti-human CD146 human mouse chimeric antibody weight are analyzed by BLAST
Chain variable region sequence has 41% similarity (including CDRs), and framework region is preferably anti-human CD146 human mouse chimeric antibody people
The framework region of source design improvement.Based on the sequence similarity of JH3minigene (Kabat Vol.II), the region germline V passes through
Computer simulation is in conjunction with FR4.The WGQG of JH3minigene residue falls in the region CDR3 of anti-human CD146 human mouse chimeric antibody
In.Based on sequence alignment and the influence to antibody function is considered, using 14 humanized heavy chains of IGHV3-33 Frame Design
Mutant.H0 is the framework region that the CDRs of anti-human CD146 human mouse chimeric antibody is grafted directly to human antibody.H1-H11 is in H0
On the basis of transformation, be mutated on 71,69,66,30,27,28,73,78,24,48,38 position respectively.H12 is in H0 base
Back mutation is carried out in 66,69 and 71 sites on plinth.H13 is on the basis of H0 above 7 27,28,30,66,69,71,73
Site carries out back mutation.H14 is that site on the basis of H0 71,69,66,30,27,28,73,78,24,48,38 carries out
Back mutation.Back mutation refers to the sequence by source of people sequence amino acid inverse transition at anti-human CD146 human mouse chimeric antibody.
The area people V gene database is analyzed by BLAST, selection can with anti-human CD146 human mouse chimeric antibody light chain
There is substitution of the people IGKV1-39 of 64% similitude (including CDRs) as the area FR in change area.Based on sequence similarity analysis, J-
The FR4 frame of 2 minigene of region kappa (Kabat Vol.II) is most suitable.The first two of JK-2 minigene is residual
Base is fallen in the anti-human area CD146 human mouse chimeric antibody CDR3.By sequence alignment and in view of issuable to antibody function
It influences, obtains 6 light chain mutants containing IGKV1-39 framework region.Structure L0 is by anti-human CD146 human mouse chimeric antibody
CDR is grafted directly to people's IGKV1-39 framework region.L1-44 structure and the difference of L0 are that they are respectively in 2,1,3 and 60 sites
It is mutated.L5 is in 4 site simultaneous mutations.
Embodiment 2 determines specificity of the antibody of humanization design in conjunction with people's CD146 receptor by ELISA
Synthesize VH structural domain and VL structural domain according to computer simulation humanization design's result, by the VH structural domain of synthesis and
VL structural domain is cloned into mammalian expression vector, which enables corresponding structural domain in complete human IgG bone
It is expressed in frame.By heavy chain construct and light chain construct cotransfection into mammalian cell, the small quantities of of humanized antibody is carried out
Amount preparation.The supernatant of transient transfection is subjected to ELISA detection.By albumin A or G resin, IgG purification to uniform, for
It is detected in measurement based on cell.
To determine specificity of the humanized antibody in conjunction with CD146, ELISA has been carried out for CD146 and reference protein.It will
The supernatant (protein concentration is about 5 μ g/ml) of above-mentioned transient transfection dilutes 20 times, 200 times, 2000 times, 20000 times and is used to connect
The ELISA detection got off.Known CD146 is diluted to 1~10 μ g/ml with coating buffer, every hole adds 0.1ml, 4 DEG C overnight into
Row coating.Next day washs 3 times.Add certain diluted humanized antibody expression product sample (above-mentioned supernatant) 0.1ml in above-mentioned
In the reacting hole being coated with, sets 37 DEG C and be incubated for 1 hour, washing.(while doing blank, feminine gender and positive hole control) is in reacting hole
In, be added enzyme mark secondary antibody (antiantibody) 0.1ml of diluted fresh, 37 DEG C incubation 30-60 minutes, washing, last is all over using
DDW washing.Add substrate solution to develop the color: being added the tmb substrate solution 0.1ml of Extemporaneous in each reacting hole, 37 DEG C 10~30 points
Clock.It terminates reaction: 2M sulfuric acid 0.05ml being added in each reacting hole.Result judgement: it directly can with the naked eye be seen in white background
Examine result: color is deeper in reacting hole, and positive degree is stronger, and negative reaction is colourless or extremely shallow, the depth of the be in color of foundation,
It is indicated with "+", "-" number.OD450 value can also be surveyed: on ELISA detector, at 450nm, being surveyed after being returned to zero with blank control wells
Each hole OD450 value, it is as positive if more than 2.1 times of defined negative control OD450 value.As a result as shown in table 1- table 4.Root
Preferred several groups of antibody are selected according to experimental result, as shown in table 5.
Table 1: supernatant dilutes 20 times of ELISA testing results
Table 2: supernatant dilutes 200 times of ELISA testing results
Antibody | OD450 | Antibody | OD450 | Antibody | OD450 | Antibody | OD450 | Antibody | OD450 | Antibody | OD450 |
H0L0 | 2.456 | H2L4 | 2.354 | H5L2 | 2.357 | H8L0 | 2.238 | H10L4 | 2.378 | H13L2 | 2.397 |
H0L1 | 2.378 | H2L5 | 2.317 | H5L3 | 2.279 | H8L1 | 2.020 | H10L5 | 2.315 | H13L3 | 2.442 |
H0L2 | 2.482 | H3L0 | 2.281 | H5L4 | 2.291 | H8L2 | 2.098 | H11L0 | 2.190 | H13L4 | 2.321 |
H0L3 | 2.469 | H3L1 | 2.315 | H5L5 | 2.241 | H8L3 | 2.070 | H11L1 | 2.326 | H13L5 | 2.315 |
H0L4 | 2.437 | H3L2 | 2.273 | H6L0 | 2.230 | H8L4 | 2.162 | H11L2 | 2.317 | H14L0 | 2.317 |
H0L5 | 2.368 | H3L3 | 2.120 | H6L1 | 2.220 | H8L5 | 1.991 | H11L3 | 2.236 | H14L1 | 2.220 |
H1L0 | 2.152 | H3L4 | 2.319 | H6L2 | 2.306 | H9L0 | 2.391 | H11L4 | 2.354 | H14L2 | 2.295 |
H1L1 | 1.844 | H3L5 | 2.418 | H6L3 | 2.408 | H9L1 | 2.456 | H11L5 | 2.485 | H14L3 | 2.467 |
H1L2 | 2.188 | H4L0 | 2.510 | H6L4 | 2.311 | H9L2 | 2.420 | H12L0 | 1.434 | H14L14 | 2.312 |
H1L3 | 2.168 | H4L1 | 2.303 | H6L5 | 2.225 | H9L3 | 2.271 | H12L1 | 1.056 | H14L15 | 2.275 |
H1L4 | 2.226 | H4L2 | 2.281 | H7L0 | 1.364 | H9L4 | 2.244 | H12L2 | 1.412 | Blank | 0.067 |
H1L5 | 1.919 | H4L3 | 2.390 | H7L1 | 2.284 | H9L5 | 2.270 | H12L3 | 1.244 | Blank | 0.063 |
H2L0 | 2.329 | H4L4 | 2.341 | H7L2 | 2.261 | H10L0 | 2.267 | H12L4 | 1.345 | blank89 | 0.061 |
H2L1 | 2.225 | H4L5 | 2.214 | H7L3 | 2.159 | H10L1 | 2.195 | H12L5 | 1.101 | blank90 | 0.054 |
H2L2 | 2.378 | H5L0 | 2.407 | H7L4 | 2.222 | H10L2 | 2.344 | H13L0 | 2.277 | blank0 | 0.048 |
H2L3 | 2.481 | H5L1 | 2.440 | H7L5 | 2.400 | H10L3 | 2.494 | H13L1 | 2.387 | blank0 | 0.051 |
Table 3: supernatant dilutes 2000 times of ELISA testing results
Table 4: supernatant dilutes 20000 times of ELISA testing results
Antibody | OD450 | Antibody | OD450 | Antibody | OD450 | Antibody | OD450 | Antibody | OD450 | Antibody | OD450 |
H0L0 | 0.277 | H2L4 | 0.333 | H5L2 | 0.278 | H8L0 | 0.180 | H10L4 | 0.280 | H13L2 | 0.263 |
H0L1 | 0.333 | H2L5 | 0.340 | H5L3 | 0.291 | H8L1 | 0.145 | H10L5 | 0.262 | H13L3 | 0.348 |
H0L2 | 0.380 | H3L0 | 0.325 | H5L4 | 0.300 | H8L2 | 0.151 | H11L0 | 0.202 | H13L4 | 0.211 |
H0L3 | 0.292 | H3L1 | 0.324 | H5L5 | 0.260 | H8L3 | 0.171 | H11L1 | 0.288 | H13L5 | 0.330 |
H0L4 | 0.333 | H3L2 | 0.298 | H6L0 | 0.182 | H8L4 | 0.195 | H11L2 | 0.396 | H14L0 | 0.478 |
H0L5 | 0.361 | H3L3 | 0.247 | H6L1 | 0.252 | H8L5 | 0.154 | H11L3 | 0.351 | H14L1 | 0.325 |
H1L0 | 0.158 | H3L4 | 0.274 | H6L2 | 0.240 | H9L0 | 0.284 | H11L4 | 0.400 | H14L2 | 0.268 |
H1L1 | 0.127 | H3L5 | 0.304 | H6L3 | 0.255 | H9L1 | 0.278 | H11L5 | 0.358 | H14L3 | 0.325 |
H1L2 | 0.167 | H4L0 | 0.435 | H6L4 | 0.225 | H9L2 | 0.332 | H12L0 | 0.091 | H14L14 | 0.317 |
H1L3 | 0.190 | H4L1 | 0.292 | H6L5 | 0.262 | H9L3 | 0.247 | H12L1 | 0.084 | H14L15 | 0.310 |
H1L4 | 0.205 | H4L2 | 0.318 | H7L0 | 0.093 | H9L4 | 0.276 | H12L2 | 0.098 | Blank | 0.065 |
H1L5 | 0.132 | H4L3 | 0.326 | H7L1 | 0.282 | H9L5 | 0.314 | H12L3 | 0.091 | Blank | 0.07 |
H2L0 | 0.295 | H4L4 | 0.333 | H7L2 | 0.296 | H10L0 | 0.246 | H12L4 | 0.091 | blank89 | 0.052 |
H2L1 | 0.251 | H4L5 | 0.243 | H7L3 | 0.287 | H10L1 | 0.277 | H12L5 | 0.093 | blank90 | 0.059 |
H2L2 | 0.336 | H5L0 | 0.312 | H7L4 | 0.212 | H10L2 | 0.324 | H13L0 | 0.383 | blank0 | 0.051 |
H2L3 | 0.397 | H5L1 | 0.279 | H7L5 | 0.275 | H10L3 | 0.285 | H13L1 | 0.273 | blank0 | 0.055 |
Table 5: the combination that ELISA result is best in the supernatant of different dilutions is selected
It is in consolidated statement 5 as a result, expression quantity preferable light and weight chain combinational expression higher to some affinity in above-mentioned experiment
Antibody carries out further ELISA verifying.Thin thereon it will release 5000 times and 20000 times progress ELISA detections respectively.As a result such as
Shown in table 6- table 7.H5L0 antibody, H10L2 antibody, H14L0 antibody, H11L3 antibody, H11L4 antibody, H13L0 antibody, H0L2 are anti-
Body, H11L2 antibody, H9L2 antibody, H13L4 antibody, H2L3 antibody, H0L4 antibody, H9L0 antibody, H2L4 antibody, H2L2 antibody,
H7L3 antibody, H0L5 antibody, H4L0 antibody, H12L0 antibody, H14L1 are currently preferred antibody.
Above-mentioned affinity is higher, the preferable light and weight chain combinational expression antibody of expression quantity, the following institute of the amino acid sequence for including
It states:
H5:SEQ ID NO.1;H10:SEQ ID NO.2;H14:SEQ ID NO.3;H11:SEQ ID NO.4;H13:
SEQ ID NO.5;H0:SEQ ID NO.6;H9:SEQ ID NO.7;H2:SEQ ID NO.8;H7:SEQ ID NO.9;H4:
SEQ ID NO.10;H12:SEQ ID NO.11;L0:SEQ ID NO.12;L2:SEQ ID NO.13;L3:SEQ ID
NO.14;L4:SEQ ID NO.15;L1:SEQ ID NO.16;L5:SEQ ID NO.17.
Nucleic acid sequence corresponding to above-mentioned amino acid sequence is as described below:
H5:SEQ ID NO.20;H10:SEQ ID NO.21;H14:SEQ ID NO.22;H11:SEQ ID NO.23;
H13:SEQ ID NO.24;H0:SEQ ID NO.25;H9:SEQ ID NO.26;H2:SEQ ID NO.27;H7:SEQ ID
NO.28;H4:SEQ ID NO.29;H12:SEQ ID NO.30;L0:SEQ ID NO.31;L2:SEQ ID NO.32;L3:SEQ
ID NO.33;L4:SEQ ID NO.34;L1:SEQ ID NO.35;L5:SEQ ID NO.36.
Table 6: the ELISA result after preferred 5000 times of supernatant dilution in conjunction with CD146
Table 7: the ELISA result after preferred 20000 times of supernatant dilution in conjunction with CD146
Antibody | OD450 value | Antibody | OD450 value | Antibody | OD450 value |
H0L2 | 0.259 | H7L3 | 0.261 | H13L0 | 0.246 |
H0L4 | 0.266 | H9L0 | 0.284 | H13L4 | 0.196 |
H0L5 | 0.284 | H9L2 | 0.219 | H14L0 | 0.242 |
H2L2 | 0.28 | H10L2 | 0.268 | H14L1 | 0.273 |
H2L3 | 0.346 | H11L2 | 0.36 | Blank | 0.056 |
H2L4 | 0.278 | H11L3 | 0.265 | Blank | 0.059 |
H4L0 | 0.281 | H11L4 | 0.316 | blank0 | 0.051 |
H5L0 | 0.254 | H12L0 | 0.081 | blank0 | 0.055 |
Embodiment 3: the combination of humanized antibody series and CD146 carry out BIACORE analysis
The purpose of the present embodiment is that the binding affinity of humanization modified antibody Yu people CD146 is analyzed with BIACORE.It is described
Humanization modified antibody be that H5L0 antibody (comprising amino acid sequence SEQIDNO1 and SEQIDNO12), H10L2 antibody (include
Amino acid sequence SEQIDNO2 and SEQIDNO13), H14L0 antibody (include amino acid sequence SEQIDNO3 and SEQIDNO12),
H11L3 antibody (including amino acid sequence SEQIDNO4 and SEQIDNO14) H11L4 antibody (includes amino acid sequence SEQIDNO4
And SEQIDNO15) H13L0 antibody (include amino acid sequence SEQIDNO5 and SEQIDNO12), H0L2 antibody (includes amino acid
Sequence SEQIDNO6 and SEQIDNO13), H11L2 antibody (include amino acid sequence SEQIDNO4 and SEQIDNO13), H9L2 it is anti-
Body (include amino acid sequence SEQIDNO7 and SEQIDNO13), H13L4 antibody (comprising amino acid sequence SEQIDNO5 and
SEQIDNO15), H2L3 antibody (including amino acid sequence SEQIDNO8 and SEQIDNO14), H0L4 antibody (include amino acid sequence
Arrange SEQIDNO6 and SEQIDNO15), H9L0 antibody (include amino acid sequence SEQIDNO7 and SEQIDNO12), H2L4 antibody
(include amino acid sequence SEQIDNO8 and SEQIDNO15), H2L2 antibody (comprising amino acid sequence SEQIDNO8 and
SEQIDNO13), H7L3 antibody (including amino acid sequence SEQIDNO9 and SEQIDNO14).
Analysis carries out on BIACORE3000 instrument, which carries out before prepared by each new chip and passed through system
Test.All operationss 25 DEG C carry out, with HBS-EP (GEHealthcareBR-1006-69/5mMHEPES, 150mMNaCl,
3.4mMEDTA, 0.005%surfactantP20, pH7.4) it is used as working buffer solution.It (resonates using by primary amine chemical property
Unit be about 4000RU ' s) target protein A of the covalent coupling on CM5 chip at similar resonating member (100-200RU ' s)
Confrontation CD146 human mouse chimeric antibody and humanized antibody are analyzed.Each chip is also prepared for i.e. not even with reference surface
The control of connection capture antibody.Obtain the induction curve figure (Sensorgrams) of the circulation carried out using different analyte concentrations with
Just dynamic analysis is carried out.One circulation by monoclonal antibody be captured on the surface, with flowing working buffer solution carry out it is short
Temporary stationary phase and subsequent limiting concentration analyte (ECD or subdomain albumen) are bonded.Injection is tied with surface
The analyte (3-4 minutes) of conjunction obtains the bound fraction in curve.Then buffer elution (3-4 minutes) are carried out, record dissociation
Data.Circulation ends here, and the actified solution (10mM glycine pH1.5) that injection capture kit provides captures to remove
Antibody/analyte, but not significantly affect capture antibody and carry out the capture of monoclonal antibody again in subsequent circulation.
The conventional method of affinity analysis is as follows: firstly, preparing chip and testing it to different MAb concentrations
The resonance units (RU) of lower capture.Then kinetic loop is executed, monoclonal antibody is captured to the water of general 100RU in circulation
It is flat, allow to analyze albumen and be combined then dissociation, surface is regenerated except the whole albumen for eliminating covalent coupling.With
6 kinds of different analysis protein concentrations (usually 200nM, 100nM, 50nM, 25nM, 12.5nM and 6.25nM) carry out it is a series of this
Kind circulation.More wheel buffer circulations are carried out before analyzing albumen to guarantee the consistency of circulation, in some tests, buffer
Circulation is by as " dual reference ".The analyte used is people CD146.In dynamic test cyclic process, analogies coupling
Surface provide reference, which is subtracted from the specific antibody being currently running-analyte RU data to remove buffer band
The illusion come.To each concentration in kinetic curve group, subtract from the data of the circulation containing analyte containing only buffer
Circulation data, thus to it is certain operation do dual reference.It will be obtained in this way using BIAEVALUATION software (v.3.2)
The curve data overall situation be fitted 11 Lang Gemiaoer (Langmuir) model.Utilize BIAEVALUATIONTM Software Create phase interaction
With induction curve figure, dynamic assessment is carried out with curve graph.Experimental result is as shown in table 8.
Table 8: anti-CD146 human mouse chimeric antibody and humanized antibody and people CD146 receptor affinity analyze result
Antibody | Analyte | Ka(1/Ms) | Kd(1/s) | KD(M) |
H5L0 | People CD146 | 7.98x104 | 6.48x10-3 | 8.11x10-8 |
H10L2 | People CD146 | 2.76x105 | 1.42x10-2 | 5.15x10-8 |
H14L0 | People CD146 | 1.62x105 | 3.30x10-3 | 2.03x10-8 |
H11L3 | People CD146 | 3.31x105 | 8.88x10-3 | 2.68x10-8 |
H11L4 | People CD146 | 9.65x104 | 7.83x10-3 | 8.11x10-8 |
H13L0 | People CD146 | 7.22x104 | 5.57x10-3 | 7.71x10-8 |
H0L2 | People CD146 | 1.13x105 | 1.06x10-2 | 9.39x10-8 |
H11L2 | People CD146 | 1.20x105 | 8.65x10-3 | 7.21x10-8 |
H9L2 | People CD146 | 1.60x105 | 9.05x10-3 | 5.65x10-8 |
H13L4 | People CD146 | 1.59x105 | 5.52x10-3 | 3.48x10-8 |
H2L3 | People CD146 | 1.51x105 | 9.70x10-3 | 6.41x10-8 |
H0L4 | People CD146 | 2.95x105 | 1.10x10-2 | 3.72x10-8 |
H9L0 | People CD146 | 1.50x105 | 8.92x10-3 | 5.95x10-8 |
H2L4 | People CD146 | 1.55x105 | 1.03x10-2 | 6.61x10-8 |
H2L2 | People CD146 | 1.58x105 | 1.00x10-2 | 6.37x10-8 |
H7L3 | People CD146 | 1.10x105 | 9.51x10-3 | 8.67x10-8 |
Anti- CD146 human mouse chimeric antibody | People CD146 | 8.34x104 | 3.28x10-3 | 3.94x10-8 |
Table 8 shows the data that BIACORE is obtained, and can be seen that the humanization designed by computer simulation from KD value
Antibody (H14L0) and humanized antibody (H11L3) are best to the affinity of people CD146, are higher than source of mouse antibody in conjunction with activity, are not
Come the good candidate antibody for developing therapeutic anti-CD146 monoclonal antibody.In above data, H13L4, H0L4, H10L2, H9L2,
The affinity and source of mouse of the monoclonal antibody and CD146 that obtain after several humanization designs of H9L0, H2L2, H2L3, H2L4 are anti-
Body is very nearly the same.It proves antibody provided by the present invention, antibody can be improved in the case where affinity is constant or even increases
Humanization degree.
It should be pointed out that for those of ordinary skill in the art, without departing from the principle of the present invention, may be used also
With several improvements and modifications are made to the present invention, these improvements and modifications also fall within the scope of protection of the claims of the present invention.
Sequence table
<110>Nanjing Ren Nuo pharmaceutcal corporation, Ltd
<120>a kind of Humanized anti-human CD146 monoclonal antibody for having efficient neutralization activity
<130> 2017
<160> 38
<170> SIPOSequenceListing 1.0
<210> 1
<211> 118
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 1
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Phe Ser Asn Tyr
20 25 30
Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Arg Ile Tyr Pro Gly Thr Asp Ile Thr Tyr Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ser Gly Gly Tyr Trp Tyr Phe Asp Val Trp Gly Ala Gly Thr
100 105 110
Met Val Thr Val Ser Ser
115
<210> 2
<211> 118
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 2
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr
20 25 30
Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile
35 40 45
Ala Arg Ile Tyr Pro Gly Thr Asp Ile Thr Tyr Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ser Gly Gly Tyr Trp Tyr Phe Asp Val Trp Gly Ala Gly Thr
100 105 110
Met Val Thr Val Ser Ser
115
<210> 3
<211> 118
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 3
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Thr Ser Gly Tyr Ile Phe Thr Asn Tyr
20 25 30
Trp Ile His Trp Val Lys Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile
35 40 45
Ala Arg Ile Tyr Pro Gly Thr Asp Ile Thr Tyr Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Lys Phe Thr Leu Ser Val Asp Lys Ser Lys Asn Thr Ala Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ser Gly Gly Tyr Trp Tyr Phe Asp Val Trp Gly Ala Gly Thr
100 105 110
Met Val Thr Val Ser Ser
115
<210> 4
<211> 118
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 4
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr
20 25 30
Trp Ile His Trp Val Lys Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Arg Ile Tyr Pro Gly Thr Asp Ile Thr Tyr Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ser Gly Gly Tyr Trp Tyr Phe Asp Val Trp Gly Ala Gly Thr
100 105 110
Met Val Thr Val Ser Ser
115
<210> 5
<211> 118
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 5
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Ile Phe Thr Asn Tyr
20 25 30
Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Arg Ile Tyr Pro Gly Thr Asp Ile Thr Tyr Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Lys Phe Thr Leu Ser Val Asp Lys Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ser Gly Gly Tyr Trp Tyr Phe Asp Val Trp Gly Ala Gly Thr
100 105 110
Met Val Thr Val Ser Ser
115
<210> 6
<211> 118
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 6
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr
20 25 30
Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Arg Ile Tyr Pro Gly Thr Asp Ile Thr Tyr Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ser Gly Gly Tyr Trp Tyr Phe Asp Val Trp Gly Ala Gly Thr
100 105 110
Met Val Thr Val Ser Ser
115
<210> 7
<211> 118
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 7
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Thr Ser Gly Phe Thr Phe Ser Asn Tyr
20 25 30
Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Arg Ile Tyr Pro Gly Thr Asp Ile Thr Tyr Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ser Gly Gly Tyr Trp Tyr Phe Asp Val Trp Gly Ala Gly Thr
100 105 110
Met Val Thr Val Ser Ser
115
<210> 8
<211> 118
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 8
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr
20 25 30
Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Arg Ile Tyr Pro Gly Thr Asp Ile Thr Tyr Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Arg Phe Thr Leu Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ser Gly Gly Tyr Trp Tyr Phe Asp Val Trp Gly Ala Gly Thr
100 105 110
Met Val Thr Val Ser Ser
115
<210> 9
<211> 118
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 9
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr
20 25 30
Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Arg Ile Tyr Pro Gly Thr Asp Ile Thr Tyr Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Lys Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ser Gly Gly Tyr Trp Tyr Phe Asp Val Trp Gly Ala Gly Thr
100 105 110
Met Val Thr Val Ser Ser
115
<210> 10
<211> 118
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 10
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Asn Tyr
20 25 30
Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Arg Ile Tyr Pro Gly Thr Asp Ile Thr Tyr Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ser Gly Gly Tyr Trp Tyr Phe Asp Val Trp Gly Ala Gly Thr
100 105 110
Met Val Thr Val Ser Ser
115
<210> 11
<211> 118
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 11
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr
20 25 30
Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Arg Ile Tyr Pro Gly Thr Asp Ile Thr Tyr Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Lys Phe Thr Leu Ser Val Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ser Gly Gly Tyr Trp Tyr Phe Asp Val Trp Gly Ala Gly Thr
100 105 110
Met Val Thr Val Ser Ser
115
<210> 12
<211> 112
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 12
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Lys Ser Val Ser Ile Ser
20 25 30
Gly Tyr Ser Tyr Met His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro
35 40 45
Lys Leu Leu Ile Tyr Leu Ala Ser Asn Leu Glu Ser Gly Val Pro Ser
50 55 60
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
65 70 75 80
Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln His Ser Arg
85 90 95
Glu Leu Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg
100 105 110
<210> 13
<211> 112
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 13
Glu Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Lys Ser Val Ser Ile Ser
20 25 30
Gly Tyr Ser Tyr Met His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro
35 40 45
Lys Leu Leu Ile Tyr Leu Ala Ser Asn Leu Glu Ser Gly Val Pro Ser
50 55 60
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
65 70 75 80
Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln His Ser Arg
85 90 95
Glu Leu Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg
100 105 110
<210> 14
<211> 112
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 14
Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Lys Ser Val Ser Ile Ser
20 25 30
Gly Tyr Ser Tyr Met His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro
35 40 45
Lys Leu Leu Ile Tyr Leu Ala Ser Asn Leu Glu Ser Gly Val Pro Ser
50 55 60
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
65 70 75 80
Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln His Ser Arg
85 90 95
Glu Leu Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg
100 105 110
<210> 15
<211> 112
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 15
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Lys Ser Val Ser Ile Ser
20 25 30
Gly Tyr Ser Tyr Met His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro
35 40 45
Lys Leu Leu Ile Tyr Leu Ala Ser Asn Leu Glu Ser Gly Val Pro Ala
50 55 60
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
65 70 75 80
Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln His Ser Arg
85 90 95
Glu Leu Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg
100 105 110
<210> 16
<211> 112
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 16
Asp Leu Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Lys Ser Val Ser Ile Ser
20 25 30
Gly Tyr Ser Tyr Met His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro
35 40 45
Lys Leu Leu Ile Tyr Leu Ala Ser Asn Leu Glu Ser Gly Val Pro Ser
50 55 60
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
65 70 75 80
Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln His Ser Arg
85 90 95
Glu Leu Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg
100 105 110
<210> 17
<211> 112
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 17
Glu Leu Val Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Lys Ser Val Ser Ile Ser
20 25 30
Gly Tyr Ser Tyr Met His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro
35 40 45
Lys Leu Leu Ile Tyr Leu Ala Ser Asn Leu Glu Ser Gly Val Pro Ala
50 55 60
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
65 70 75 80
Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln His Ser Arg
85 90 95
Glu Leu Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg
100 105 110
<210> 18
<211> 330
<212> PRT
<213>homo sapiens (homo sapiens)
<400> 18
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys
1 5 10 15
Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr
20 25 30
Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
35 40 45
Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser
50 55 60
Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr
65 70 75 80
Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys
85 90 95
Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys
100 105 110
Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
115 120 125
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
130 135 140
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
145 150 155 160
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
165 170 175
Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
180 185 190
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn
195 200 205
Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
210 215 220
Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu
225 230 235 240
Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr
245 250 255
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn
260 265 270
Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe
275 280 285
Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn
290 295 300
Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
305 310 315 320
Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
325 330
<210> 19
<211> 107
<212> PRT
<213>homo sapiens (homo sapiens)
<400> 19
Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu
1 5 10 15
Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe
20 25 30
Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln
35 40 45
Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser
50 55 60
Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu
65 70 75 80
Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser
85 90 95
Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
100 105
<210> 20
<211> 354
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 20
caggtgcagc tggtggagtc cggcggcggc gtggtgcagc ccggccgctc cctgcgcctg 60
tcctgcgccg cctccggcta caccttctcc aactactgga tccactgggt gcgccaggcc 120
cccggcaagg gcctggagtg ggtggcccgc atctaccccg gcaccgacat cacctactac 180
aacgagaagt tcaagggccg cttcaccatc tcccgcgaca actccaagaa caccctgtac 240
ctgcagatga actccctgcg cgccgaggac accgccgtgt actactgcgc ccgctccggc 300
ggctactggt acttcgacgt gtggggcgcc ggcaccatgg tgaccgtgtc ctcc 354
<210> 21
<211> 354
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 21
caggtgcagc tggtggagtc cggcggcggc gtggtgcagc ccggccgctc cctgcgcctg 60
tcctgcgccg cctccggctt caccttctcc aactactgga tccactgggt gcgccaggcc 120
cccggcaagg gcctggagtg gatcgcccgc atctaccccg gcaccgacat cacctactac 180
aacgagaagt tcaagggccg cttcaccatc tcccgcgaca actccaagaa caccctgtac 240
ctgcagatga actccctgcg cgccgaggac accgccgtgt actactgcgc ccgctccggc 300
ggctactggt acttcgacgt gtggggcgcc ggcaccatgg tgaccgtgtc ctcc 354
<210> 22
<211> 354
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 22
caggtgcagc tggtggagtc cggcggcggc gtggtgcagc ccggccgctc cctgcgcctg 60
tcctgcgcca cctccggcta catcttcacc aactactgga tccactgggt gaagcaggcc 120
cccggcaagg gcctggagtg gatcgcccgc atctaccccg gcaccgacat cacctactac 180
aacgagaagt tcaagggcaa gttcaccctg tccgtggaca agtccaagaa caccgcctac 240
ctgcagatga actccctgcg cgccgaggac accgccgtgt actactgcgc ccgctccggc 300
ggctactggt acttcgacgt gtggggcgcc ggcaccatgg tgaccgtgtc ctcc 354
<210> 23
<211> 354
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 23
caggtgcagc tggtggagtc cggcggcggc gtggtgcagc ccggccgctc cctgcgcctg 60
tcctgcgccg cctccggctt caccttctcc aactactgga tccactgggt gaagcaggcc 120
cccggcaagg gcctggagtg ggtggcccgc atctaccccg gcaccgacat cacctactac 180
aacgagaagt tcaagggccg cttcaccatc tcccgcgaca actccaagaa caccctgtac 240
ctgcagatga actccctgcg cgccgaggac accgccgtgt actactgcgc ccgctccggc 300
ggctactggt acttcgacgt gtggggcgcc ggcaccatgg tgaccgtgtc ctcc 354
<210> 24
<211> 354
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 24
caggtgcagc tggtggagtc cggcggcggc gtggtgcagc ccggccgctc cctgcgcctg 60
tcctgcgccg cctccggcta catcttcacc aactactgga tccactgggt gcgccaggcc 120
cccggcaagg gcctggagtg ggtggcccgc atctaccccg gcaccgacat cacctactac 180
aacgagaagt tcaagggcaa gttcaccctg tccgtggaca agtccaagaa caccctgtac 240
ctgcagatga actccctgcg cgccgaggac accgccgtgt actactgcgc ccgctccggc 300
ggctactggt acttcgacgt gtggggcgcc ggcaccatgg tgaccgtgtc ctcc 354
<210> 25
<211> 354
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 25
caggtgcagc tggtggagtc cggcggcggc gtggtgcagc ccggccgctc cctgcgcctg 60
tcctgcgccg cctccggctt caccttctcc aactactgga tccactgggt gcgccaggcc 120
cccggcaagg gcctggagtg ggtggcccgc atctaccccg gcaccgacat cacctactac 180
aacgagaagt tcaagggccg cttcaccatc tcccgcgaca actccaagaa caccctgtac 240
ctgcagatga actccctgcg cgccgaggac accgccgtgt actactgcgc ccgctccggc 300
ggctactggt acttcgacgt gtggggcgcc ggcaccatgg tgaccgtgtc ctcc 354
<210> 26
<211> 354
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 26
caggtgcagc tggtggagtc cggcggcggc gtggtgcagc ccggccgctc cctgcgcctg 60
tcctgcgcca cctccggctt caccttctcc aactactgga tccactgggt gcgccaggcc 120
cccggcaagg gcctggagtg ggtggcccgc atctaccccg gcaccgacat cacctactac 180
aacgagaagt tcaagggccg cttcaccatc tcccgcgaca actccaagaa caccctgtac 240
ctgcagatga actccctgcg cgccgaggac accgccgtgt actactgcgc ccgctccggc 300
ggctactggt acttcgacgt gtggggcgcc ggcaccatgg tgaccgtgtc ctcc 354
<210> 27
<211> 354
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 27
caggtgcagc tggtggagtc cggcggcggc gtggtgcagc ccggccgctc cctgcgcctg 60
tcctgcgccg cctccggctt caccttctcc aactactgga tccactgggt gcgccaggcc 120
cccggcaagg gcctggagtg ggtggcccgc atctaccccg gcaccgacat cacctactac 180
aacgagaagt tcaagggccg cttcaccctg tcccgcgaca actccaagaa caccctgtac 240
ctgcagatga actccctgcg cgccgaggac accgccgtgt actactgcgc ccgctccggc 300
ggctactggt acttcgacgt gtggggcgcc ggcaccatgg tgaccgtgtc ctcc 354
<210> 28
<211> 354
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 28
caggtgcagc tggtggagtc cggcggcggc gtggtgcagc ccggccgctc cctgcgcctg 60
tcctgcgccg cctccggctt caccttctcc aactactgga tccactgggt gcgccaggcc 120
cccggcaagg gcctggagtg ggtggcccgc atctaccccg gcaccgacat cacctactac 180
aacgagaagt tcaagggccg cttcaccatc tcccgcgaca agtccaagaa caccctgtac 240
ctgcagatga actccctgcg cgccgaggac accgccgtgt actactgcgc ccgctccggc 300
ggctactggt acttcgacgt gtggggcgcc ggcaccatgg tgaccgtgtc ctcc 354
<210> 29
<211> 354
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 29
caggtgcagc tggtggagtc cggcggcggc gtggtgcagc ccggccgctc cctgcgcctg 60
tcctgcgccg cctccggctt caccttcacc aactactgga tccactgggt gcgccaggcc 120
cccggcaagg gcctggagtg ggtggcccgc atctaccccg gcaccgacat cacctactac 180
aacgagaagt tcaagggccg cttcaccatc tcccgcgaca actccaagaa caccctgtac 240
ctgcagatga actccctgcg cgccgaggac accgccgtgt actactgcgc ccgctccggc 300
ggctactggt acttcgacgt gtggggcgcc ggcaccatgg tgaccgtgtc ctcc 354
<210> 30
<211> 354
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 30
caggtgcagc tggtggagtc cggcggcggc gtggtgcagc ccggccgctc cctgcgcctg 60
tcctgcgccg cctccggctt caccttctcc aactactgga tccactgggt gcgccaggcc 120
cccggcaagg gcctggagtg ggtggcccgc atctaccccg gcaccgacat cacctactac 180
aacgagaagt tcaagggccg cttcaccctg tccgtggaca actccaagaa caccctgtac 240
ctgcagatga actccctgcg cgccgaggac accgccgtgt actactgcgc ccgctccggc 300
ggctactggt acttcgacgt gtggggcgcc ggcaccatgg tgaccgtgtc ctcc 354
<210> 31
<211> 336
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 31
gacatccaga tgacccagtc cccctcctcc ctgtccgcct ccgtgggcga ccgcgtgacc 60
atcacctgcc gcgcctccaa gtccgtgtcc atctccggct actcctacat gcactggtac 120
cagcagaagc ccggcaaggc ccccaagctg ctgatctacc tggcctccaa cctggagtcc 180
ggcgtgccct cccgcttctc cggctccggc tccggcaccg acttcaccct gaccatctcc 240
tccctgcagc ccgaggactt cgccacctac tactgccagc actcccgcga gctgccctac 300
accttcggcc agggcaccaa gctggagatc aagcgc 336
<210> 32
<211> 336
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 32
gagatccaga tgacccagtc cccctcctcc ctgtccgcct ccgtgggcga ccgcgtgacc 60
atcacctgcc gcgcctccaa gtccgtgtcc atctccggct actcctacat gcactggtac 120
cagcagaagc ccggcaaggc ccccaagctg ctgatctacc tggcctccaa cctggagtcc 180
ggcgtgccct cccgcttctc cggctccggc tccggcaccg acttcaccct gaccatctcc 240
tccctgcagc ccgaggactt cgccacctac tactgccagc actcccgcga gctgccctac 300
accttcggcc agggcaccaa gctggagatc aagcgc 336
<210> 33
<211> 336
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 33
gacatcgtga tgacccagtc cccctcctcc ctgtccgcct ccgtgggcga ccgcgtgacc 60
atcacctgcc gcgcctccaa gtccgtgtcc atctccggct actcctacat gcactggtac 120
cagcagaagc ccggcaaggc ccccaagctg ctgatctacc tggcctccaa cctggagtcc 180
ggcgtgccct cccgcttctc cggctccggc tccggcaccg acttcaccct gaccatctcc 240
tccctgcagc ccgaggactt cgccacctac tactgccagc actcccgcga gctgccctac 300
accttcggcc agggcaccaa gctggagatc aagcgc 336
<210> 34
<211> 336
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 34
gacatccaga tgacccagtc cccctcctcc ctgtccgcct ccgtgggcga ccgcgtgacc 60
atcacctgcc gcgcctccaa gtccgtgtcc atctccggct actcctacat gcactggtac 120
cagcagaagc ccggcaaggc ccccaagctg ctgatctacc tggcctccaa cctggagtcc 180
ggcgtgcccg cccgcttctc cggctccggc tccggcaccg acttcaccct gaccatctcc 240
tccctgcagc ccgaggactt cgccacctac tactgccagc actcccgcga gctgccctac 300
accttcggcc agggcaccaa gctggagatc aagcgc 336
<210> 35
<211> 336
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 35
gacctgcaga tgacccagtc cccctcctcc ctgtccgcct ccgtgggcga ccgcgtgacc 60
atcacctgcc gcgcctccaa gtccgtgtcc atctccggct actcctacat gcactggtac 120
cagcagaagc ccggcaaggc ccccaagctg ctgatctacc tggcctccaa cctggagtcc 180
ggcgtgccct cccgcttctc cggctccggc tccggcaccg acttcaccct gaccatctcc 240
tccctgcagc ccgaggactt cgccacctac tactgccagc actcccgcga gctgccctac 300
accttcggcc agggcaccaa gctggagatc aagcgc 336
<210> 36
<211> 336
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 36
gagctggtga tgacccagtc cccctcctcc ctgtccgcct ccgtgggcga ccgcgtgacc 60
atcacctgcc gcgcctccaa gtccgtgtcc atctccggct actcctacat gcactggtac 120
cagcagaagc ccggcaaggc ccccaagctg ctgatctacc tggcctccaa cctggagtcc 180
ggcgtgcccg cccgcttctc cggctccggc tccggcaccg acttcaccct gaccatctcc 240
tccctgcagc ccgaggactt cgccacctac tactgccagc actcccgcga gctgccctac 300
accttcggcc agggcaccaa gctggagatc aagcgc 336
<210> 37
<211> 990
<212> DNA
<213>homo sapiens (homo sapiens)
<400> 37
gcctccacca agggcccctc cgtgttcccc ctggccccct cctccaagtc cacctccggc 60
ggcaccgccg ccctgggctg cctggtgaag gactacttcc ccgagcccgt gaccgtgtcc 120
tggaactccg gcgccctgac ctccggcgtg cacaccttcc ccgccgtgct gcagtcctcc 180
ggcctgtact ccctgtcctc cgtggtgacc gtgccctcct cctccctggg cacccagacc 240
tacatctgca acgtgaacca caagccctcc aacaccaagg tggacaagaa ggtggagccc 300
aagtcctgcg acaagaccca cacctgcccc ccctgccccg cccccgagct gctgggcggc 360
ccctccgtgt tcctgttccc ccccaagccc aaggacaccc tgatgatctc ccgcaccccc 420
gaggtgacct gcgtggtggt ggacgtgtcc cacgaggacc ccgaggtgaa gttcaactgg 480
tacgtggacg gcgtggaggt gcacaacgcc aagaccaagc cccgcgagga gcagtacaac 540
tccacctacc gcgtggtgtc cgtgctgacc gtgctgcacc aggactggct gaacggcaag 600
gagtacaagt gcaaggtgtc caacaaggcc ctgcccgccc ccatcgagaa gaccatctcc 660
aaggccaagg gccagccccg cgagccccag gtgtacaccc tgcccccctc ccgcgacgag 720
ctgaccaaga accaggtgtc cctgacctgc ctggtgaagg gcttctaccc ctccgacatc 780
gccgtggagt gggagtccaa cggccagccc gagaacaact acaagaccac cccccccgtg 840
ctggactccg acggctcctt cttcctgtac tccaagctga ccgtggacaa gtcccgctgg 900
cagcagggca acgtgttctc ctgctccgtg atgcacgagg ccctgcacaa ccactacacc 960
cagaagtccc tgtccctgtc ccccggcaag 990
<210> 38
<211> 320
<212> DNA
<213>homo sapiens (homo sapiens)
<400> 38
cgcaccgtgg ccgccccctc cgtgttcatc ttccccccct ccgacgagca gctgaagtcc 60
ggcaccgcct ccgtggtgtg cctgctgaac aacttctacc cccgcgaggc caaggtgcag 120
tggaaggtgg acaacgccct gcagtccggc actcccagga gtccgtgacc gagcaggact 180
ccaaggactc cacctactcc ctgtcctcca ccctgaccct gtccaaggcc gactacgaga 240
agcacaaggt gtacgcctgc gaggtgaccc accagggcct gtcctccccc gtgaccaagt 300
ccttcaaccg cggcgagtgc 320
Claims (7)
1. a kind of Humanized anti-human CD146 monoclonal antibody, it is characterised in that: the monoclonal antibody include light chain variable region and
Heavy chain variable region: where the amino acid sequence of light chain variable region is SEQ ID NO.12, and the amino acid sequence of heavy chain variable region is
SEQ ID NO.3;Or, the amino acid sequence of light chain variable region is SEQ ID NO.14, the amino acid sequence of heavy chain variable region is
SEQ ID NO.4;Or, the amino acid sequence of light chain variable region is SEQ ID NO.15, the amino acid sequence of heavy chain variable region is
SEQ ID NO.5;Or, the amino acid sequence of light chain variable region is SEQ ID NO.15, the amino acid sequence of heavy chain variable region is
SEQ ID NO.6;Or, the amino acid sequence of light chain variable region is SEQ ID NO.13, the amino acid sequence of heavy chain variable region is
SEQ ID NO.2;Or, the amino acid sequence of light chain variable region is SEQ ID NO.13, the amino acid sequence of heavy chain variable region is
SEQ ID NO.7;Or, the amino acid sequence of light chain variable region is SEQ ID NO.12, the amino acid sequence of heavy chain variable region is
SEQ ID NO.7;Or, the amino acid sequence of light chain variable region is SEQ ID NO.13, the amino acid sequence of heavy chain variable region is
SEQ ID NO.8;Or, the amino acid sequence of light chain variable region is SEQ ID NO.14, the amino acid sequence of heavy chain variable region is
SEQ ID NO.8;Or, the amino acid sequence of light chain variable region is SEQ ID NO.15, the amino acid sequence of heavy chain variable region is
SEQ ID NO.8。
2. monoclonal antibody according to claim 1, it is characterised in that: the monoclonal antibody further includes heavy chain constant region
And constant region of light chain.
3. monoclonal antibody according to claim 2, it is characterised in that: the amino acid sequence of the heavy chain constant region is
Amino acid sequence shown in SEQIDNO:18.
4. monoclonal antibody according to claim 2, it is characterised in that: the amino acid sequence of the constant region of light chain is
Amino acid sequence shown in SEQIDNO:19.
5. any Humanized anti-human CD146 monoclonal antibody is in preparation for treating malignant tumour in claim 1-4
Or the application in Autoimmune diseases or new vessels class disease medicament.
6. a kind of for treating the pharmaceutical composition of tumour, it includes any Humanized anti-human CD146 of such as claim 1-5 is mono-
Clonal antibody and pharmaceutical carrier.
7. a kind of expression vector, the expression vector includes coding: the nucleic acid sequence of SEQ ID NO.22, SEQ ID NO.31;
Or, including the nucleic acid sequence of SEQ ID NO.23, SEQ ID NO.33;Or, including SEQ ID NO.24, SEQ ID NO.34
Nucleic acid sequence;Or, including the nucleic acid sequence of SEQ ID NO.25, SEQ ID NO.34;Or, including SEQ ID NO.21, SEQ
The nucleic acid sequence of ID NO.32;Or, including the nucleic acid sequence of SEQ ID NO.26, SEQ ID NO.32;Or, including SEQ ID
The nucleic acid sequence of NO.26, SEQ ID NO.31;Or, including the nucleic acid sequence of SEQ ID NO.27, SEQ ID NO.32;Or, packet
Include the nucleic acid sequence of SEQ ID NO.27, SEQ ID NO.33;Or, including the nucleic acid of SEQ ID NO.27, SEQ ID NO.34
Sequence.
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PCT/CN2019/070428 WO2019137309A1 (en) | 2018-01-09 | 2019-01-04 | Humanized anti-human cd146 monoclonal antibody having high neutralizing activity |
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CN108129567B (en) * | 2018-01-09 | 2019-02-26 | 南京任诺药业有限公司 | A kind of Humanized anti-human CD146 monoclonal antibody having efficient neutralization activity |
WO2023133595A2 (en) | 2022-01-10 | 2023-07-13 | Sana Biotechnology, Inc. | Methods of ex vivo dosing and administration of lipid particles or viral vectors and related systems and uses |
WO2024040194A1 (en) | 2022-08-17 | 2024-02-22 | Capstan Therapeutics, Inc. | Conditioning for in vivo immune cell engineering |
WO2024081820A1 (en) | 2022-10-13 | 2024-04-18 | Sana Biotechnology, Inc. | Viral particles targeting hematopoietic stem cells |
WO2024119157A1 (en) | 2022-12-02 | 2024-06-06 | Sana Biotechnology, Inc. | Lipid particles with cofusogens and methods of producing and using the same |
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CN102936283B (en) * | 2008-01-31 | 2014-11-05 | 中国科学院生物物理研究所 | Anti-human CD146 monoclonal antibodies, compositions containing anti-human CD146 monoclonal antibodies, and soluble CD146 detection method |
WO2015044218A1 (en) * | 2013-09-24 | 2015-04-02 | Oncoinvent As | Monoclonal antibody and derivatives |
CN104650231A (en) * | 2015-02-13 | 2015-05-27 | 南京任诺药业有限公司 | Human-derived anti-human CD146 monoclonal antibody with efficient neutralizing activity |
WO2017046774A2 (en) * | 2015-09-16 | 2017-03-23 | Prothena Biosciences Limited | Use of anti-mcam antibodies for treatment or prophylaxis of giant cell arteritis, polymyalgia rheumatica or takayasu's arteritis |
CN106589124B (en) * | 2016-11-29 | 2019-03-15 | 中国人民解放军陆军军医大学第一附属医院 | CD146 monoclonal antibody detects and separates the application in identification in glioma perivascular cell |
CN108129567B (en) * | 2018-01-09 | 2019-02-26 | 南京任诺药业有限公司 | A kind of Humanized anti-human CD146 monoclonal antibody having efficient neutralization activity |
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