WO1989007657A1 - Precurseurs de proteines amyloides, sondes genetiques, anticorps, et modes d'utilisation - Google Patents

Precurseurs de proteines amyloides, sondes genetiques, anticorps, et modes d'utilisation Download PDF

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WO1989007657A1
WO1989007657A1 PCT/US1989/000549 US8900549W WO8907657A1 WO 1989007657 A1 WO1989007657 A1 WO 1989007657A1 US 8900549 W US8900549 W US 8900549W WO 8907657 A1 WO8907657 A1 WO 8907657A1
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app
nucleic acid
sequence
antibody
exon
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PCT/US1989/000549
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Rachael L. Neve
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The Children's Medical Center Corporation
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
    • G01N33/6896—Neurological disorders, e.g. Alzheimer's disease
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
    • C07K14/4711—Alzheimer's disease; Amyloid plaque core protein
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/81—Protease inhibitors
    • C07K14/8107—Endopeptidase (E.C. 3.4.21-99) inhibitors
    • C07K14/811—Serine protease (E.C. 3.4.21) inhibitors
    • C07K14/8114—Kunitz type inhibitors
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/38—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against protease inhibitors of peptide structure
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/158—Expression markers

Definitions

  • This invention relates to the general field of - detecting and investigating Alzheimer's disease (AD) and Down Syndrome (DS) .
  • AD Alzheimer's disease
  • DS Down Syndrome
  • the term Alzheimer's disease describes a progressive neurodegene ive disorder characterized by gradual loss of memory, reasoning, orientation, and judgment. It has been reported that a proportion of Alzheimer's disease cases are caused y a genetic defect that is transmitted in an autosomal dominant fashion and has been mapped by genetic linkage analysis to chromosome 21. St. George-Hyslop et al. (1987) Science 235: 885-890.
  • AD Alzheimer's disease
  • DS Down Syndrome
  • trisomy 21 is characterized by an extra copy of chromosome 21.
  • Neuritic plaques, neurofibrillary tangles, and ⁇ 5 cerebrovascular deposits containing AP are associated with DS, as well as AD.
  • APP human amyloid protein precursor
  • APP-1 The key distinguishing feature between APP-1 and APP-2 is the exon of APP-1, which is located so as to interrupt and alter the Val 289 encoding codon of APP-2, as described more fully below.
  • APP-2 is apparently the protein encoded by the clones described in Goldgaber et al . , Kang et al. , Robokis et al . , and Tanzi et al .
  • nucleic acid probes comprising a nucleic acid sequence capable of differentiating between APP-1 and APP-2 nucleic acid by selective hybridization.
  • differential hybridization I mean the probes will hybridize to only one form of APP nucleic acid under appropriately stringent conditions.
  • the probes which selectively hybridizes to APP-2 and not to APP-1, comprises a DNA sequence consisting of an APP-2 nucleic acid sequence spanning the APP-2 Val 289-encoding codon (i.e., a sequence that includes the Val-289 codon and subsegments on either 5 side of it that are too short to hybridize to APP-1 nucleic acid) .
  • the APP-2 DNA sequence of the featured probe in its entirely, is long enough to hybridize to APP-2 nucleic acid (particularly to APP-2 mRNA), but it does not extend far enough on either side of the Val 289 Q encoding codon to hybridize to APP-1 nucleic acid, and it lacks sequences that will hybridize to the APP-1 exon nucleic acid. In that way, one preferred probe will hybridize to APP-2 mRNA, but not to APP-1 mRNA.
  • the preferred probe includes the Val 289 ⁇ codon of APP-2 and subsegments of up to about 20 (e.g. 8-22) bases on either side of that codon.
  • Another preferred probe can hybridize to APP-1 nucleic acid but not to APP-2 nucleic acid, by virtue of inclusion in the probe of a sequence from the APP-1 exon long enough to 0 hybridize to APP-1 nucleic acid.
  • a second aspect of the invention includes differentiating APP-1 and APP-2 nucleic acid using the probes of the first aspect in an assay which comprises contacting a sample with the probes under conditions which allow differential hybridization and detecting that hybridization.
  • Differential hybridization means hybridization under sufficiently stringent conditions (e.g., 37°C, 50% formamide, 5XSSC) to hybridize to the target but not to other nucleic acid in the sample.
  • a DNA probe is contacted with mRNA, most preferably brain or neuronal mRNA, squadand the hybridization is detected by a Northern blot analysis.
  • a third aspect of the invention generally features a substantially purified and substantially isolated polypeptide comprising the exon-encoded amino acid sequence, or a polypeptide that is immunologically cross-reactive with the exon-encoded amino acid sequence.
  • substantially purified and substantially isolated means that the polypeptide has been separated from the components of the environment in which it naturally occurs to enable the use of the polypeptide as described below.
  • polypeptide broadly to include proteins as well as smaller polypeptides. Two polypeptides are
  • immunologically cross-reactive if both peptides bind to an single specific antibody, i.e., an antibody that does not substantially bind to other compenents in the environment of use.
  • polypeptides are immunologically cross reactive if each includes a binding domain (epitope) that will bind to the same specific monoclonal antibody.
  • binding domain epitope
  • two proteins having a common antigenic determinant at least 6-10 amino acid residues long can be immunologically cross-reactive.
  • One preferred polypeptide is an APP-1 cross-reactive polypeptide that comprises an amino acid sequence that is substantially identical to the sequence of the APP-1 exon or an antigenic fragment (e.g. a sequence of at least eight amino acid residues) of the exon.
  • the APP-1 cross-reactive polypeptide lacks any determinants that cross-react with APP-2; -for example, the polypeptide lacks any APP-1 sequences outside the exon that are greater than six amino acids in length.
  • Another preferred polypeptide is APP-2 cross-reactive , and it spans the exon junction to create a binding domain that does not cross-react with APP-l (e.g. -from E285 to A293.
  • the APP-2 cross-reactive peptide lacks sufficient length (it has less than six residues on either side of position 289) to cross-react with APP-l.
  • a fourth aspect of the invention generally features antibodies that react with, and/or are raised by challenge -with, the above described polypeptides.
  • the invention features antibodies which are capable of discriminating between APP-l and APP-2.
  • a mammal is challenged with one of the above-described peptide fragments of APP-l or APP-2 and discriminating antibodies are selected.
  • the mammal is challenged with the peptide fragments, and clonal antibody-producing cells are obtained from the mammal, from which an APP-l or APP-2 specific antibody-producing clone is selected.
  • the antibodies are useful in methods of assaying for the above-described abnormalities in which a sample is contacted with the antibody and an immunological reaction with the antibody is detected.
  • a fifth aspect of the invention generally features an expression vehicle comprising a DNA sequence encoding one of the polypeptides of the first aspect of the invention.
  • Fig. 1 shows the cDNA sequence of a segment of clone HL124 containing the APP sequence beginning 3 ' to bp 865 extending through the APP-l exon.
  • Fig. 2 is the APP cDNA sequence reported by Kang et al . Nature 325_: 733-736 (1987).
  • Fig. 3A is a Northern blot analysis of mRNA from various normal human fetal tissue using: a) FB68L (Tanzi et al. Science, cited above) which hybridizes to mRNA of APP-l and APP-2; b) HL124i, which hybridizes only to APP-l mRNA; and c) AMY3 , which hybridizes only to APP-2 mRNA.
  • Fig. 3B is a Northern blot analysis of mRNA from various normal adult brain tissue using the three probes described above.
  • 0 Fig. 4 is a Northern blot analysis of mRNA from normal, AD, and DS brain tissue using the three probes discussed above, as well as a control probe for tau protein.
  • Fig. 5 is a Northern blot analysis of an adult 5 Down Syndrome brain using the three probes described above. APP-l and APP-2
  • the present invention takes advantage of the existence of the two different forms of a precursor to Q amyloid ⁇ -protein (APP-l and APP-2) expressed in brain and peripheral tissue, and the different pattern of expression of APP-l and APP-2 in tissue. Specifically, we have found that patients suffering from AD and DS exhibit a different pattern of APP-l/APP-2 expression from that of normal individuals.
  • APP-l and APP-2 Q amyloid ⁇ -protein
  • APP-l and APP-2 are differentiated by the existence in APP-l of an exon.
  • the exon of interest is contained in the APP cDNA sequence of Fig. 1.
  • Fig. 2 shows the APP-2 cDNA sequence reported by Kang et al . Nature 325 : 733-736 (1987).
  • nucleotide residues are numbered in the 5' to 3 ' direction, beginning with the first base of the initiation codon AUG.
  • the untranslated sequence directly following the poly(G) tail is indicated by negative numbers.
  • the sequence shows a 695-residue open reading frame.
  • the deduced a ino-acid sequence is numbered, and the amino-terminal methionine is included.
  • the amino-acid sequence of the A4 polypeptide is boxed. Zigzag underline shows a probable membrane- spanning sequence of the A4 polypeptide.
  • the synthetic oligonucleotide mixture used as probe is indicated as a line above the corresponding cDNA. Polyadenylation signals are underlined. Nucleotide 3207 is followed by a poly(dA) tail linked to the vector DNA.
  • the exon is the 168 bp of DNA corresponding between bp 865 and bp 866 of APP-l. This extra segment interrupts the Val 289 codon of APP-2, changing it to lie. The exon preserves the reading frame, adding 56 amino acids to the N-terminus of that newly created lie.
  • the exon is shown to map to the same region of chromosome 21 as the APP sequence, by hybridizing a TaqI - HALII segment of the exon to various DNA, including chromosome 21.
  • the Taql-Haell segment detects a BamI restriction fragment length polymorphism that shows close genetics linkage to two EcoRI RFLP' s detected by fetal brain APP cDNAs, FB68L and FB130 (a 5' clone of FB68L) . See Tanzi et al . , cited above, which is hereby incorporated by reference.
  • Fig. 1 the dark triangle denotes the positions of potential splice junction sites in HL124i. Sequences flanking the 3' and 5' ends of HL124i are underlined. A 22-bp region to which an antisense oligonucleotide (denoted HL124i), was constructed for Northern blot analysis described below is overlined. The Taq I and Hae II sites are shown. Assays and Probes
  • cDNA probes can be used to evaluate the mRNA j - transcripts in tissue to determine the effect of exon on AP expression and degradation.
  • One group of useful probes in particular includes cDNA probes which hybridize to APP-2 but not to APP-l. Such probes can be constructed by selecting a
  • suitable probe as described above would include sequences with no more than about 22 bases on either side ' ⁇ ' f the Val 289 codon.
  • Suitable probe sequences can be synthesized by standard DNA synthesis techniques, using the sequence given in Fig. 2. For example, an
  • Another group of useful probes are those which contain a sequence from the exon of Fig. 1 long enough to hybridize to APP-2 nucleic acid. A sequence of at least 15 and preferably at least 20-35 is preferred. These sequences also can be synthesized by known techniques.
  • the preferred assays using these probes involve hybridization of a DNA probe sequence to sample mRNA using conventional Northern Blot techniques.
  • a DNA probe sequence to sample mRNA using conventional Northern Blot techniques.
  • levels of APP-2 mRNA in brain and neuronal tissue are directly correlated to AD and DS disease.
  • Northern blot analysis of mRNA from such tissue using a probe specific for APP-2 mRNA is useful to evaluate patients for such disease.
  • AD and DS involves a general decrease of APP-2 expression in affected areas of the brain, but a substantial increase in APP-l expression in brain and neuronal tissue. Accordingly, it is also useful to assay specifically for APP-l and for APP-2 to follow their relative expression levels. Such assays can be accomplished with cDNA probes specific for APP-l mRNA.
  • the overlined sequence of Fig. 1 (HL124i) can be used to derive an antisense sequence useful to hybridize specifically to APP-l mRNA.
  • a sequence spanning the Val 289-encoding codon (AMY3) is useful to hybridize specifically to APP-2 mRNA.
  • a cDNA library was constructed by standard techniques (see Neve et al . (1986) Mol . Brain Res . 1 :271-280) from mRNA of the human promyelocytic leukemia cell line HL60, available from Dr. Stuart Orkin, at The Childrens Medical Center Corporation, 300 Longwood Avenue, Boston, MA 02115.
  • the library was screened with FB68L, a fetal brain cDNA corresponding to the 3' portion of the APP gene.
  • Fig. 1 depicts a restriction fragment map of FB68L, as well as . sequence information from that cDNA and probes used to locate that cDNA.
  • Nucleic acid probes according to the present invention are -capable of selectively hybridizing to the exon portion of the APP-l mRNA, to detect over expression of that gene.
  • a DNA probe comprising at least 15 (and preferably at least 20-35) bases of that exon sequence can be used to hybridize to mRNA of various tissue.
  • the probes can be used to detect APP transcripts (mRNA) or cDNA derived from them, using standard Norther blot techniques. Over expression of APP transcripts in neuronal tissues is indicative of DS. Selective .loss of APP-2 transcript from the frontal cortex is indicative of AD. The normal pattern exhibits higher APP-2 expression in the fetal brain, and a similar pattern of expression in adult cortex.
  • Fig. 3A is comparison of -FB68L, HL124i and AMY3 expressio -in-human 20-22 week tissues. Fetal tissue was obtained from midtrimester elective abortuses under protocols approved by the institutional review board at Brigham and Women's Hospital. The blot to which HL124i and AMY3 were hybridized was equivalent to that used in the FB68L hybridization.
  • the 22 bp oligonucleotide termed HL124i is homologous to a relatively non-conserved portion of the nucleotide sequence encoding the protease inhibitor domain present in the previously described alternate APP message (see Fig. 1).
  • the sequence of HL124i is 5 'CATGCAGTACTCTTCTGTGTCA3 '
  • the 40 bp oligonucleotide termed AMY3 encompasses 20 bp on either side of the potential splice junction sites in the cDNA HL124 (see Fig. 2) which includes the protease inhibitor domain; it encompasses 40 contiguous bases in cDNAs lacking this domain.
  • the sequence of AMY3 is 5 'CTGGCTGCTGTTGTAGGAACTCGAACCACCTTTCCACAGA3 ' . Hybridizations with HL124i were carried out in 5XSSC,
  • RNA isolation and hybridization with FB68L have previously been described by Tanzi et al. , cited above.
  • AMY3 hybridizes to RNA species of the same molecular weight, but hybridization is only seen in human fetal brain. The apparent positive result for AMY3 in kidney tissue is believed to represent non-specific hybridization to an overload line.
  • Fig. 3B is a comparison of HL124i and AMY3 expression in 18 week fetal tissue.
  • HL124i and AMY3 were successively hybridized to the same RNA blot.
  • the protocol used is the same as generally described for Example 1.
  • These hybridizations confirmed the ubiquitous nature of the transcript detected by HL124i, and added meninges, spinal cord, and cerebellum to the tissues in which this RNA was shown to be expressed.
  • the AMY3 transcript was additionally detected in spinal cord and cerebellum but not in any other tissue including meninges. Note the lack of positive hybridization to kidney RNA in this panel.
  • the RNA to which AMY3 hybridizes that is, the RNA lacking the protease inhibitor domain, appears to be expressed selectively in the nervous system.
  • FIG. 4 shows Northern blots of FB68L, HL124i and AMY3 hybridizations to total mRNA (25 micrograms) from 19-week normal (lane a) and trisomy 21 (lane b) brains, adult normal (lane c) and AD (lane d) cerebellum, and adult normal (lane e) and AD (lane f) frontal cortex.
  • Fetal tissue was obtained from an abortus with a diagnosis of Down Syndrome and from an age-matched normal abortus.
  • Adult tissue was obtained from autopsy brains of a case of histologically confirmed AD and from an individual without dementing illness.
  • Fig. 5- shows hybridizations of FB68L, HL124i and AMY3 to RNA (10 ug) from the brain of a 37 yr. Down Syndrome female (23.6 hours post mortem).
  • Abbreviations are as follows: A6, supplementary motor cortex; A7, parietal cortex; A19, extrastriate cortex; Al, primary somatosensory cortex; A10, frotal pole of the cortex; A17, striate cortex; A18, extrastriate cortex; A20, 21, temporal association cortex; A4, motor cortex; cb, cerebellum; C-P, caudate-putamen; Th, thalamus-ventral posterolateral nucleus; Hi, hippocampus; A40, posterior perisylvian cortex-supra marginal gyrus; A44, anterior perisylvian cortex-opercular gyrus.
  • the brain was confirmed upon autopsy to display the neuropathological characteristics of aged Down Syndrom brains (neuritic plagues
  • AMY3 transcript is expressed normally at higher levels than is that of HL124i in the 19-week fetal brain.
  • the intensity of HL124i and AMY3 hybridizations to mRNA from a 19-week Down Syndrome fetal brain are both increased several-fold relative to hybridizations with RNA from the normal 19-week fetal brain, indicating that both APP transcripts are overexpressed in Down Syndrome brain.
  • Hybridizations of HL124i and AMY3 to adult normal and AD cerebellum show that both transcripts are present at approximately normal levels in AD cerebellum.
  • HL124i and AMY3 hybridizations to normal and AD frontal cortex reveal striking differences in the expression of the two transcripts.
  • the level of HL124i mRNA is near normal, while the AMY3 transcript appears to be selectively lost in AD frontal cortex.
  • the patterns of expression of AMY3 and HL124i transcripts shown here were confirmed in two additional cases of fetal DS brain relative, .to age-matched controls and in two additional cases of AD cerebellum and cortex compared to adult controls.
  • Example 4 was repeated on a brain from a 57 year old Down Syndrome female (13.2 hours post mortem) yielding similar patterms of hybridization (not shown) .
  • Example 6 For comparison with Examples 4 and 5, FB68L, HLl24i, and AMY3 probes were hybridized to RNAs from different regions of a control adult human brain (82 year old male) .
  • Hybridization with FB68L shows marked regional variation in the adult human brain. Expression of the " gene is highest in the association cortex, specifically in Brodman areas (A) 10, A20/21, A40, and A4 . In ' contrast, the distribution of the HL124i transcript is relatively homogenous across the brain regions. The differential expression of APP in the brain revealed by FB68L is apparently primarily due to the APP-2 transcript. The stronger signal in associative areas of the neocortex seen upon hybridization with FB68L indicates a relative enrichment in these regions of the APP-2 (detected by AMY3). Notably, APP-l mRNA is expressed at considerably higher levels in the hippocampus than is the mRNA detected by AMY3. o Those skilled in the filed will also understand that the above probes and assays are useful not only in clinical diagnosis, but also in autopsy to evaluate and confirm diagnosis, and in clinical research to explore the molecular basis for Alzheimer Disease and Down 5 Syndrome.
  • HL124i could be modified in-length e.g. to include more than 22 0 bases; such an alternative having 39 bases has the following sequence:
  • AMY3 could be modified in length; such an alternative having 22 bases has the following sequence:
  • Polypeptides according to the invention include APP-2, which has the amino acid sequence inferred from the APP-2 cDNA reported by Kang et al . , (see Fig. 2), except for the exon discussed above.
  • the invention also includes the exon-encoded peptide of APP-l. Conservative substitutions can be made in the exon-encoded peptide, or in the exon-encoded fragment of APP-l without departing from the spirit of the invention.
  • immunologically cross-reactive fragments of the exon encoded sequence are within the scope of the invention.
  • Polypeptides according to the invention can be obtained by expression of the cDNA sequence of Fig.
  • Particular APP-1-specific antigens include polypeptides according to the inventions include those which comprise a hapten sequence of at least eight and preferably more amino acids from the exon.
  • Particular APP-2-specific polypeptides include a hapten sequence of at least eight amino acids spanning the Val-289 codon of APP-2. These haptens sequences are coupled to a carrier to provide a molecule of sufficient size to raise an immune response, according to well known techniques.
  • Polyclonal antibodies can be recovered by standard techniques and used to separate APP-l from APP-2 with an immuno ffinity column. Monoclonal antibodies can be recovered by generating a population of hybridomas using standard techniques, and screening for cell lines producing antibodies specific to APP-l or APP-2.
  • Immunoassays for APP—1 and/or APP-2 are performed using the resulting antibodies in any of a variety of known immunoassay formats, e.g. radioimmunoassay.

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Abstract

On peut, au moyen de sondes d'acides nucléiques et de techniques d'analyse d'hybridation, détecter et différencier deux formes différentes de précurseur de protéines amyloïdes (PPA). Les deux formes de PPA diffèrent par l'existence d'un exon dans une forme présentant une homologie avec des inhibiteurs de protéase de sérine. On peut utiliser des antigènes de protéine afin de développer des anticorps distinguant le PPA-1 du PPA-2 dans une immuno-analyse. La distinction entre le PPA-1 et le PPA-2 est utile pour déterminer l'évolution de la maladie d'Alzheimer et du syndrome de Down.
PCT/US1989/000549 1988-02-10 1989-02-10 Precurseurs de proteines amyloides, sondes genetiques, anticorps, et modes d'utilisation WO1989007657A1 (fr)

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Cited By (18)

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WO1991004339A1 (fr) * 1989-09-18 1991-04-04 California Biotechnology Inc. Techniques d'analyse et reactifs servant a detecter les depots d'amyloide
WO1991015773A1 (fr) * 1990-04-07 1991-10-17 Oxford Virology Plc Procede de test diagnostique
GB2256642A (en) * 1991-06-13 1992-12-16 Ici Plc Nucleotide sequences for detecting alzheimer's disease
US5221607A (en) * 1989-09-18 1993-06-22 Scios Nova Inc. Assays and reagents for amyloid deposition
EP0561087A1 (fr) * 1992-03-20 1993-09-22 N.V. Innogenetics S.A. Forme mutée du gène de la protéine du précurseur beta-amyloide
WO1994021683A1 (fr) * 1992-03-20 1994-09-29 N.V. Innogenetics S.A. FORME MUTANTE DU GENE DE PROTEINE PRECURSEUR DE β-AMYLOIDE
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US9175094B2 (en) 2007-06-12 2015-11-03 Ac Immune S.A. Monoclonal antibody
US9176150B2 (en) 2003-01-31 2015-11-03 AbbVie Deutschland GmbH & Co. KG Amyloid beta(1-42) oligomers, derivatives thereof and antibodies thereto, methods of preparation thereof and use thereof
US9221900B2 (en) 2010-07-30 2015-12-29 Ac Immune S.A. Methods for identifying safe and functional humanized antibodies
US9403902B2 (en) 2007-10-05 2016-08-02 Ac Immune S.A. Methods of treating ocular disease associated with amyloid-beta-related pathology using an anti-amyloid-beta antibody
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