EP1131345A1 - Polypeptides synthetiques correspondant au virus de l'hepatite c (hcv) et applications - Google Patents

Polypeptides synthetiques correspondant au virus de l'hepatite c (hcv) et applications

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Publication number
EP1131345A1
EP1131345A1 EP99956280A EP99956280A EP1131345A1 EP 1131345 A1 EP1131345 A1 EP 1131345A1 EP 99956280 A EP99956280 A EP 99956280A EP 99956280 A EP99956280 A EP 99956280A EP 1131345 A1 EP1131345 A1 EP 1131345A1
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EP
European Patent Office
Prior art keywords
amino acid
seq
polypeptide according
peptide
different
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Application number
EP99956280A
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German (de)
English (en)
Inventor
Pascal Dalbon
Michel Jolivet
Colette Jolivet-Reynaud
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Biomerieux SA
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Biomerieux SA
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Publication of EP1131345A1 publication Critical patent/EP1131345A1/fr
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Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
    • C12N2770/00011—Details
    • C12N2770/24011—Flaviviridae
    • C12N2770/24211—Hepacivirus, e.g. hepatitis C virus, hepatitis G virus
    • C12N2770/24222—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes

Definitions

  • the present invention generally relates to 5 synthetic polypeptides, that is to say which are obtained by preparative routes such as chemical synthesis, composed of consecutive amino acids which are together identical to any fragment, sequence or region of the structural protein of the nucleocapsid called CORE protein of the human 0 hepatitis C virus (HCV) .
  • CORE protein of the human 0 hepatitis C virus HCV
  • These polypeptides can be used as synthetic antigens in various applications arising from their immunogenicity and which are specified below; at the forefront of these applications is the detection HCV in various body fluids such as for example a blood sample.
  • nucleocapsid protein or CORE protein of HCV which is composed as established by Figure 1 of 191 amino acids (SEQ ID NO:3), is that which exhibits the greatest homology, on the one hand, between the sequences of the same group of viral 0 isolates, and, on the other hand, between the different groups of viruses.
  • this CORE protein is encoded by a structural part of the HCV genome and therefore constitutes a structural protein. The high conservation of the structure of this protein makes it a particularly 5 suitable candidate for the immunological detection of HCV.
  • the one called VIIIE consisting of the sequence of N-terminal amino acids 2 to 62 of the CORE protein, was tested in an ELISA test with respect to its immunoreactivity towards the anti -HCV antibodies contained in sera from individuals infected with HCV.
  • This polypeptide demonstrated a good immunoreactivity towards the HCV- infected sera tested.
  • the inventors have tried to shorten the length of the polypeptide VIIIE by respectively amputating 9, 19, 29 and 39 amino acids from its N-terminal part in order to prepare the polypeptides consisting of the N-terminal amino acid sequences of the CORE protein with a length of 10 to 62, 20 to 62, 30 to 62 and 40 to 62 respectively.
  • the present invention provides a polypeptide, or its fragments, which although consisting of an amino acid sequence much shorter than that of the VIIIE polypeptide structure manifests an immunoreactivity with all the sera of individuals or samples infected with HCV and which carry antibodies directed against the nucleocapsid protein.
  • this immunodominant region must be continuous if it is desired to react with all the sera of individuals or blood samples infected with HCV and which possess antibodies directed against the CORE protein;
  • this immunodominant region clearly contains conformational type epitopes and linear type epitopes.
  • polypeptide used in conformity with the invention comprise-s an isolated peptide sequence which is composed of about the 45 N-terminal amino acids of the HCV virus CORE protein (cf SEQ ID NOl) .
  • the polypeptide of the invention consists of only or of an isolated peptide sequence composed of the 45 N-terminal amino acids of the said protein or alternatively of any homologous polypeptide comprising about 45 amino acids and exhibiting an anti- genic reactivity towards HCV.
  • polypeptide of the invention consists of a peptide sequence which is composed of the
  • N-terminal amino acids 2 to 45 of the CORE protein (cf SEQ ID N02) .
  • EP-A-0 569 309 relates to an isolated peptide sequence which is composed of 45 N-terminal amino acids (SEQ ID NO:l) of the CORE (or capsid) protein of the human hepatitis C virus (HCV) , as represented in Figure 1, 1 to 10 amino acids being optionally amputated from this sequence in its N-terminal part and/or its C-terminal part.
  • isolated peptide sequence is understood to mean any polypeptide not fused with another protein or another peptide regardless of its route of production, for example by chemical synthesis, by lysis of the CORE protein, or by genetic recombination techniques. This polypeptide can therefore be a synthetic peptide or a protein.
  • the preferred amputations of the polypeptide of the invention are the amputations of respectively 6 amino acids and 11 amino acids from the N-terminal end of the CORE protein.
  • the present invention concerns a polypeptide that specifically binds to antibodies that specifically bind to human hepatitis C virus consisting of an antigenic sequence having specific immunoreactivity to the antibodies of SEQ ID NO:l, with the proviso that said polypeptide is different from SEQ ID NO :1, SEQ ID NO :2 and sequences resulting from the amputation of SEQ ID NO :1 with 1 to 10 amino acids from the N-terminal part and/or the C-terminal part of SEQ ID NO :1.
  • polypeptide sequences according to the invention may be such as in the native state or modified chemically.
  • Chemical modification is understood to mean any chemical alteration of at least one functional group of the peptide sequence which essentially preserves or even develops the biological properties of the said sequence.
  • the replacement of an amino acid of the L series by an amino acid of the D series, a modification of the side chains of the amino acids such as an acetylation of the amine functional groups, a carboxymethylation of the thiol functional groups or an esterification of the carboxylic functional groups, or a modification of the peptide bonds such as carba, retro-inverso, reduced and methylene-oxy bonds, are especially part of the chemical modifications considered above.
  • a preferred polypeptide (b) comprises or consists of an antigenic sequence which consists of
  • SEQ ID N0:1 modified with at least one modification selected from the group consisting of :
  • a preferred chemical modification is at least one member selected from the group consisting of :
  • Said modification of side chains may be an acetylation of amine functional groups, a carboxymethylation of thiol functional groups or an esterification of carboxylic functional groups.
  • Said modification of peptide bonds may be forming carba, retro-inverso, reduced and methylene-oxy bonds.
  • an amino acid is said to be homologous to another amino acid when their chemical characteristics, such as polarity, hydrophobicity and/or basicity and/or acidity and/or neutrality, are essentially the same.
  • a classification based on the polarity of the side chains plits up amino acids into four groups : - non-polar or hydrophobic amino acids comprising : alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophane, methionine, which are considered as homologous amino acids, within the meaning of the above definition,
  • - amino acids having a non-charged side chain comprising : serine, threonine, tyrosine, asparagine, glutamine, cysteine, glycine, which are considered as homologous amino acids, within the meaning of the above definition, - amino acids having a negative-charged side chain
  • amino acids comprising aspartic acid and glutamic acid, which are considered as homologous amino acids, within the meaning of the above definition,
  • - amino acids having a positive-charged side chain comprising lysine, arginine, histidine, which are considered as homologous amino acids, within the meaning of the above definition ; depending on the pH, this group may further comprise asparagine and glutamine,
  • asparagine and aspartic acid can be considered as homologous amino acids, and glutamine and glutamic acid, as well.
  • Another preferred polypeptide is shorter than SEQ ID NO:l, and shares at least one, preferably at least two identical amino acid with SEQ ID NO: 5.
  • said polypeptide shares at least two identical amino acids with SEQ ID NO : 6 and/or at least one identical amino acid with SEQ ID NO: 7.
  • Said polypeptide may further have at least one amino acid which is homologous (as defined above) to at least one amino acid of SEQ ID NO: 1.
  • Said polypeptide may further have at least one amino acid which differs from at least one amino acid of SEQ ID N0:1 by a chemical modification (as defined above) of the side chain of said amino acid.
  • a preferred polypeptide is selected from the group consisting of SEQ ID NO: 8 through SEQ ID NO: 18.
  • the inventors have settled a screening test to determine whether a given polypeptide is an antigenic equivalent of polypeptide (a) or not. This test can be carried out by the one skilled in the art with routine experimentation and is described in details at the end of the description ( ⁇ Antigenic polypeptide different from but equivalent to peptide (a) ) .
  • the given polypeptide is screened against antibodies, at least one relevant antibody anti-HCV and one irrelevant antibody used as a negative control, using the usual methods to indicate the presence or absence of an antibody reaction.
  • these methods include enzyme linked immunosorbent assay (ELISA) .
  • ELISA enzyme linked immunosorbent assay
  • Suitable antibodies can either be purchased as commercially available antisera or prepared in a suitable host in accordance with well known procedures.
  • the results can be expressed as the mean OD obtained with the relevant antibody - OD obtained with the irrelevant antibody.
  • a threshold value is determined to select antigenic peptides.
  • the immunoreactivity of the selected peptides is tested with anti-HCV core positive human sera and with sera from healthy individuals.
  • the results are expressed as the mean OD obtained with the pool of anti-core positive human sera - OD anti-core negative human sera.
  • the invention provides a reagent for the detection of human hepatitis C virus (HCV) comprising as reactive substance any one of the abovementioned compounds or compositions and optionally any additive immunocompatible with the detection of HCV.
  • HCV human hepatitis C virus
  • the detection can be carried out using a polypeptide identical to those of the present invention or an antigenic equivalent peptide thereof, with optionally one anti-human immunoglobulin antibody, labelled with any conventional marker such as a radioactive, fluorescent or enzymatic marker or the like.
  • a reagent can be used both in a homogenous phase, for example in immunoprecipitation assays, and in a heterogenous phase, for example in immunoadsorption assays.
  • any suitable means of detection of HCV can be obtained, whether a detection kit or any other equivalent system or unit .
  • the abovementioned reagent is supported on a solid support immunocompatible with the reagent as a whole; in particular, the solid support is, without limitation, in the form of a microtiter plate, a sheet, a cone, a well, a bead or any other appropriate micro- particulate substrate.
  • the term solid support as used here includes all materials upon which the polypeptides according to the invention can be immobilized.
  • polysaccharides such as cellulose materials, for example paper, cellulose derivatives such as nitrocellulose and cellulose acetate; polymers such as vinyl chloride, polyethylene, polystyrene, polyacrylate or copolymers such as vinyl chloride and propylene polymer, vinyl chloride and vinyl acetate polymer; styrene-based copolymers; natural fibers such as cotton and synthetic fibers such as nylon.
  • cellulose materials for example paper, cellulose derivatives such as nitrocellulose and cellulose acetate
  • polymers such as vinyl chloride, polyethylene, polystyrene, polyacrylate or copolymers such as vinyl chloride and propylene polymer, vinyl chloride and vinyl acetate polymer
  • styrene-based copolymers such as natural fibers such as cotton and synthetic fibers such as nylon.
  • the solid support is a polystyrene polymer, a butadiene-styrene copolymer or a butadiene- styrene copolymer mixed with one or more polymers or copolymers chosen from polystyrene, styrene-acrylonitrile or styrene-methyl methylmethacrylate copolymers, polypropylenes, polycarbonates or analogs.
  • anti-HCV antibodies can be detected in any body part or fluid such as a blood sample of an individual suspected of being infected with HCV.
  • this body part and the above-mentioned reagent simply have to be brought into contact under predetermined conditions, for example of temperature, which permit an immunological reaction where appropriate, and to then detect the presence of an immune complex with this reagent.
  • Body part is understood to mean any fluid, tissue or organ of an individual, comprising or capable of comprising anti-HCV antibodies. These body parts may be a blood, plasma or serum sample or various secretions and the like.
  • any detection device or apparatus comprising a vessel for bringing the body part analyzed into contact with a reagent as defined above, and this with means which create conditions, such as temperature, - favorable for an immunological reaction where appropriate.
  • this device comprises means, especially optical, for the detection of the immune complex obtained with the reagent .
  • Another way of detecting the HCV virus using the polypeptides according to the present invention is to obtain monoclonal or polyclonal antibodies by any method known per se comprising an immunological reaction between a human or animal organism and an immunogenic agent consisting of a polypeptide composition as defined above.
  • the antibodies thus obtained can be used to detect HCV or to monitor the progression of the virus in a patient suffering from hepatitis C.
  • each of the polypeptide compositions according to the invention may constitute the active ingredient of an active immunotherapeutic composition, being optionally conjugated with an immunologically suitable support.
  • a pharmaceutically acceptable excipient may supplement the said composition.
  • Such a composition is for example a vaccinal preparation.
  • the immunodominant character of the peptide sequence according to the present invention was demonstrated in conformity with the following experimental procedure .
  • the strategy chosen consists in synthesizing long polypeptide fragments of about 40 amino acids, in the N-terminal part of the CORE protein, which belong to the sequence of about the first 120 amino acids.
  • the peptides were chemically synthesized by solid phase synthesis according to the Merrifield technique (Barany G, and Merrifield R.B, 1980, In the Peptides, 2, 1-284, Gross E and Meienhofer J, Eds Academic Press, New York) . The practical details are those described below. Peptide synthesis The peptides are synthesized on a phenyl- acetamidomethy1 (PAM) /polystyrene/divinylbenzene resin
  • the amino acids are coupled in the form of esters of hydroxy- benzotriazole (HOBT) .
  • HOBT hydroxy- benzotriazole
  • the amino acids used are obtained from Novabiochem (La ⁇ flerlfingen, Switzerland) or from
  • NMP hydrofluoric acid
  • peptidylresin For 1 g of peptidylresin, 10 ml of HF, 1 ml of anisole and 1 ml of dimethyl sulfide (DMS) are used, and the mixture is stirred for 45 minutes at -2°C. The HF is then evaporated under vacuum. After intensive washes with ether, the peptide is eluted from the resin with 10% acetic acid and then the peptide is freeze-dried.
  • HF dimethyl sulfide
  • the peptides are purified by preparative high- performance liquid chromatography on a . type C18 VYDAC column (250 x 21 mm) (The Separation Group, Hesperia, CA, USA) .
  • the elution is performed with an acetonitrile gradient at a flow rate of 22 ml/min.
  • the fractions collected are controlled by elution under isocratic conditions on an analytical C18 VYDAC column
  • the purified peptides are analyzed with the objective of assessing their amino acid composition using an automatic Applied Biosystems 420 H amino acid analyzer. Measurement of the chemical molecular mass (mean) of the peptides is obtained using the L.S.I.M.S. mass spectrometer in a positive ion mode, on a dual focusing instrument VG. ZAB.ZSEQ linked to a DEC-VAX 2000 acquisition system (VG analytical Ltd, Manchester,
  • HCV hepatitis C virus
  • Detection of anti-HCV antibodies by ELISA The wells of a microtiter plate of "NUNC maxisorb" trademark are saturated with 100 ⁇ l of a solution containing the peptide or a mixture of peptides (10 ⁇ g/ml) for 2 hours at 37°C. The plate is then emptied, then washed with a wash buffer containing 0.05% Tween 20. The wells are saturated with 100 ⁇ l of wash buffer supplemented with 10% goat serum (v/v) , then incubated for 30 minutes at 37°C, then washed again as above. The sera to be analyzed are diluted to the appropriate dilution with saturation buffer. The incubation of the sera is 1 hour at 37 °C.
  • R40R is measured by ELISA on HCV-positive sera (P 1 to P 20 and B 1 to B 16) and on normal sera (SN 10, 11, 16, 17, 18, 19) .
  • the different peptides are adsorbed on the microplates at a concentration of 10 ⁇ g/ml and the sera are used at 1/100 dilution.
  • each serum was, in a first instance, tested at the dilution 1/100.
  • the response proved saturating (value 2500) for all the peptides (example: serum P 5) a 1/1000 dilution, and if necessary a 1/10,000 dilution, was carried out.
  • the sera P6, P9 , P10, P13 , P19, P23, P24 are sera which do not possess antibodies against the CORE protein of HCV.
  • the plates used are polystyrene plates irradiated with gamma rays, which bind the peptides in a noncovalent manner via electrostatic type bonds but also hydrophobic bonds. It is possible that peptides, depending on their sequence, are selectively adsorbed, thus favoring a well defined part and thus preventing immunogenic reactivity towards another part which may have become less acceptable.
  • the inhibition experiments were carried out by reaction, in liquid phase, of the HCV sera with the peptides followed by reaction of the remaining antibodies with the peptide adsorbed onto the microplates.
  • the inhibitory peptides are incubated at a concentration of 0.1 mg/ml with sera of appropriate dilution. The rest of the manipulation is identical to that described for the ELISA test.
  • Peptide S18D, or V22G, or a mixture of both is preincubated overnight in the presence of the serum to be tested.
  • the antibodies can bind onto the corresponding sites.
  • the mixture (peptide + serum) is then incubated with the peptide S42G adsorbed onto the microtiter plates. If all the antibodies reacted during the incubation with the peptides S18D, V22G, or with the mixture, no reactivity will be observed, which will result in a 100% inhibition. In contrast, if antibodies specific for peptide S42G remain, they will then be able to react.
  • a control is carried out by preincubating each serum with peptide S42G, which makes it possible to calculate the percentage inhibition.
  • Table 3 collates the results of the inhibition of the binding of anti-HCV antibodies (dilution 1/10,000) onto peptide S42G, by preincubation of HCV sera with peptide S42G, peptide S18D, peptide S22G and the mixture of peptides S18D + S22G.
  • this experiment proves that antibodies specific for peptide S42G exist which do not react with either peptide S18D or with peptide V22G, the sum of both representing the total sequence of peptide S42G.
  • a final hypothesis to be evaluated consists in verifying that the antibodies specific for peptide S42G were not directed against the central part of peptide S42G, that is to say at the junction of peptides S18D and V22G.
  • a peptide was therefore prepared (cf Figure 1) whose sequence comprises the C-terminal part (6 amino acids) of peptide S18D and the N-terminal part (6 amino acids) of peptide V22G. 19 A
  • the first 45 amino acids are the most reactive towards HCV-positive sera.
  • the first 21 amino acids (peptide S18D) react, which shows the presence of one or more antigenic determinants on this peptide.
  • amino acids 22 to 45 also carry one or more epitopes.
  • the junction of these two sequences is also reactive .
  • sequence 1-45 of the CORE protein is pluriepitopic .
  • one or more antigenic determinants exist which are reactive only insofar as the entire sequence 2-45 is available and not in a discontinuous manner (peptides S18D + V22G) .
  • These epitopes, which are specific to peptide S42G, are without any doubt conformational type epitopes which can exist only insofar as this sequence of 44 amino acids (peptide S42G) has a suitable structure, a structure which is not obtained with smaller- sized peptides.
  • amino acid sequence of peptide S42G should not be discontinuous in order to preserve all the epi- topes, it can be asked if the N- and/or C-terminal parts of peptide S42G are involved in the epitopic conformations of S42G or carry the epitopes themselves.
  • the reactivity of the five peptides towards sera of individuals infected with HCV was evaluated in ELISA tests in conformity with the procedure described above for measuring the activity of the peptides S42G, P42Y and R40R.
  • Table 4 collates the results obtained for the peptides S42G, S32Y, P27Y, K22Y in order to examine the influence of an amputation of the N-terminal part and the C-terminal part of the peptide S42G. These results are expressed in optical density values read at 492 nm multiplied by a factor of 10 3 . 22
  • the sera P23 and P10 are sera which do not possess antibodies against the CORE protein of HCV. From this Table, it can be deduced that when the 23
  • Table 5 collates the results of the tests of immunoreactivity of the peptides S42G, P37G, K32G, during ELISA tests, to examine the influence of amputation of the N-terminal part of peptide S42G.
  • CORE protein C22-3 obtained in the form of a fusion protein with human superoxide dismutase and expressed by a yeast.
  • the sensitivity of peptide S42G is 100% relative to the CORE protein (C22-3) of the 2nd generation CHIRON RIBA HCV test.
  • the CORE protein can be replaced by the peptide S42G for ' the serological detection of HCV.
  • CAP-B a protein comprising the sequence 69-120 of the CORE protein, called CAP-B, exhibits no reactivity towards these same sera. This last result is in relative contradiction with those of the present invention since the peptide R40R which comprises the sequence 75-116 of this same protein reacts nevertheless with some sera (about 10%, cf Table 1).
  • the peptides obtained by chemical synthesis comprise only the sequence mentioned for each of them, and as explained above, this is one of the advantages linked to these synthetic peptides.
  • the recombinant proteins obtained by NASOFF et al. they are fusion proteins in which 308 amino acids, which are completely foreign to the CORE protein, are present in the N-terminal position.
  • the recombinant proteins which comprise the sequences 1-20 and 21-40 of the CORE protein are therefore composed of a foreign sequence by more than 90%.
  • the fact that the N-terminal part of the sequences 1-20 or 21-40 of the CORE protein is linked to the C-terminal part of the fusion protein contributes towards restricting the accessibility of this N-terminal region.
  • the C-terminal part is for its part detected. It is highly probable that it is these structural stresses, imposed by the production of a recombinant fusion protein in which the immunogenic part (the CORE part) represents the minor part of the fusion protein (less than 10% in this case), which lead to results contrary to those presented.
  • the complete experi- mental procedure described above clearly demonstrates, by the types of antigen-antibody reaction carried out either in solid phase (direct ELISA, cf Table 2), or by inhibition (cf Table 3), that the sequence 2-45 of the CORE protein of the HCV virus obtained by solid phase chemical synthesis not only proves substantially greater than smaller sequences (S18D or V22G) , but also that it exhibits a sensitivity equivalent to the CORE protein itself (protein C22-3 of the 2nd generation CHIRON RIBA HCV test) , and that consequently the synthetic peptide S42G can be used in serological diagnostic tests in place of the CORE protein.
  • peptide S42G appears to be the minimum but sufficient structure which, from the point of view of its antigenic properties, is equivalent to the CORE protein in its entirety and can therefore replace it in a reagent for the detection of 2 9 HCV .
  • Antigenic polypeptide different from but equivalent to peptide (a) Structural analysis and molecular modeling of the peptide S42G allowed the characterization of a tridimensional motif that is composed of 2 a-helix separated by a loop. This structure could define a conformation-dependent antigenic domain.
  • a dodecapeptide library displayed on phage (Ph.D.-12TM Phage Display Peptide Library Kit, New England BioLabs Inc ) with the mouse monoclonal antibody (Mab) 19D9D6. This Mab was directed against a conformational epitope present within this region and also recognized by human sera.
  • the immunoreactivity of phage clones was tested according to the manufacturer manual. The results were expressed as the mean OD obtained with the MAb 19D9D6 against 2.5 x 10 phages - the OD obtained with the anti borrelia burgdorferi OSPA MAb against the same number of phages .
  • the immunoreactivity of phage clones was tested according to the manufacturer manual. The results were expressed as the mean OD obtained with the pool of anti core positive human sera against 2.5 x 10 phages - the OD obtained with the pool of anti core negative human sera against the same number of phages. As shown in table 8, compared to the response obtained with negative sera, 5 out of the 6 tested clones gave a positive signal with the pool of positive sera. This results are in agreement with previous results, indicating that the epitope recognized by MAb 19D9D6 was also recognized by human sera.

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Abstract

Cette invention concerne un polypeptide qui se lie spécifiquement à des anticorps, lesquels se lient spécifiquement au virus de l'hépatite C humain. Il consiste en une séquence antigénique présentant une immunoréactivité spécifique aux anticorps de SEQ ID NO:1, sous la réserve que ledit polypeptide soit différent de SEQ ID NO:1, SEQ ID NO:2 et des séquences résultant de l'amputation de SEQ ID NO:1 de 1 à 10 acides aminés de la partie N-terminal et/ou de la partie C-terminal de SEQ ID NO:1. Des polypeptides préférés sont sélectionnés dans le groupe composé de SEQ ID NO:8 à SEQ ID NO:18.
EP99956280A 1998-11-20 1999-11-19 Polypeptides synthetiques correspondant au virus de l'hepatite c (hcv) et applications Withdrawn EP1131345A1 (fr)

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