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Claims (32)
A PATENTNI ZAHTEVKI 1. DNA polinukleotid, ki kodira polipeptid, kjer polipeptid obsega zaporedje najmanj 10 sosednjih amino kislin, ki so kodirane z genomom virusa hepatitisa C (HCV) in vsebuje antigensko determinanto, kjer je HCV značilen po tem, da (i) ima pozitivne RNA genome; (ii) omenjeni genom vsebuje odprto mesto čitanja (ORF - open reading frame), ki kodira poliprotein; in (iii) omenjeni poliprotein vsebuje zaporedje amino kislin, ki najmanj 40 % ustreza zaporedju amino kislin 859 na sliki 14.A DNA polynucleotide encoding a polypeptide, wherein the polypeptide comprises a sequence of at least 10 contiguous amino acids encoded by the hepatitis C virus (HCV) genome and contains an antigenic determinant wherein HCV is characterized in that (i) it has positive RNA genomes; (ii) said genome contains an open reading frame (ORF) encoding a polyprotein; and (iii) said polyprotein contains an amino acid sequence that corresponds to at least 40% of amino acid sequence 859 in Figure 14.2. DNA nukleotid po zahtevku 1, značilen po tem, da omenjeni poliprotein obsega zaporedje sosednjih amino kislin, ki najmanj 60 % ustreza zaporedju amino kislin 859 na sliki 14.The DNA nucleotide of claim 1, wherein said polyprotein comprises a sequence of contiguous amino acids that corresponds to at least 60% of the sequence of amino acids 859 in Figure 14.3. DNA polinukleotid po zahtevku 1 ali 2, značilen po tem, da omenjeno sosednje zaporedje kodira vsaj 15 amino kislin.A DNA polynucleotide according to claim 1 or 2, characterized in that said adjacent sequence encodes at least 15 amino acids.4. DNA polinukleotid po kateremkoli zahtevku od 1 do 3, značilen po tem, da se omenjeno sosednje zaporedje nahaja na sliki 14.DNA polynucleotide according to any one of claims 1 to 3, characterized in that said adjacent sequence is located in Figure 14.5. DNA polinukleotid po kateremkoli zahtevku od 1 do 3, značilen po tem, da je omenjeno sosednje zaporedje kodirano znotraj Iambda-gt11 cDNA knjižnice, ki je deponirana pri American Type Culture Collection (ATTC -Ameriška zbirka tipskih kultur; Op. prev.) pod številko 40394.DNA polynucleotide according to any one of claims 1 to 3, characterized in that said adjacent sequence is encoded within the Iambda-gt11 cDNA library deposited with the American Type Culture Collection (ATTC). under number 40394.6. DNA polinukleotid po kateremkoli zahtevku od 1 do 3, značilen po tem, da se omenjeno sosednje zaporedje nahaja na sliki 47.A DNA polynucleotide according to any one of claims 1 to 3, characterized in that said adjacent sequence is located in Figure 47.7. Rekombinantni vektor značilen po tem, da obsega kodirno zaporedje, ki vsebuje DNA polinukleotid po kateremkoli izmed zahtevkov od 1 do 6.A recombinant vector comprising a coding sequence comprising a DNA polynucleotide according to any one of claims 1 to 6.8. Celica gostitelja, ki je transformirana z rekombinantnim vektorjem po zahtevku 7, značilna po tem, da je kodirno zaporedje funkcionalno povezano s kontrolnim zaporedjem, ki zagotavlja ekspresijo kodirnega zaporedja preko celice gostitelja.A host cell transformed with a recombinant vector according to claim 7, characterized in that the coding sequence is operably linked to a control sequence that ensures the expression of the coding sequence via the host cell.9. Postopek proizvodnje rekombinantnega HCV polipeptida, značilen po tem, da obsega inkubacijo celice gostitelja po zahtevku 8 pri pogojih, ki zagotavljajo ekspresijo kodirnega zaporedja.A method of producing a recombinant HCV polypeptide, characterized in that it comprises incubating the host cell according to claim 8 under conditions that provide expression of the coding sequence.10. Polipeptid v v bistvu izolirani obliki, ki obsega zaporedje najmanj 10 sosednjih amino kislin, ki so kodirane z genomom virusa hepatitisa C (HCV) in ki vsebuje antigensko determinanto, kjer je HCV značilen po tem, da L (i) ima pozitivne RNA genome; (ii) omenjeni genom vsebuje odprto mesto čitanja (ORF - open reading frame), ki kodira poliprotein; in (iii) omenjeni poliprotein vsebuje zaporedje amino kislin, ki najmanj 40 % ustreza zaporedju amino kislin 859 na sliki 14.A polypeptide in substantially isolated form comprising a sequence of at least 10 contiguous amino acids encoded by the hepatitis C virus (HCV) genome and comprising an antigenic determinant wherein HCV is characterized by L (i) having positive RNA genomes ; (ii) said genome contains an open reading frame (ORF) encoding a polyprotein; and (iii) said polyprotein contains an amino acid sequence that corresponds to at least 40% of amino acid sequence 859 in Figure 14.11. Polipeptid po zahtevku 10, značilen po tem, da omenjeni poliprotein obsega zaporedje amino kislin, ki najmanj 60 % ustreza zaporedju amino kislin 859 na sliki 14.The polypeptide of claim 10, wherein said polyprotein comprises an amino acid sequence that at least 60% corresponds to the amino acid sequence 859 of Figure 14.12. Polipeptid po zahtevkih 10 ali 11, značilen po tem, da obsega najmanj 15 amino kislin.Polypeptide according to Claims 10 or 11, characterized in that it comprises at least 15 amino acids.13. Polipeptid po kateremkoli izmed zahtevkov od 10 do 12, značilen po tem, da je pripravljen s pomočjo ekspresije rekombinantne DNA.A polypeptide according to any one of claims 10 to 12, characterized in that it is prepared by the expression of recombinant DNA.14. Polipeptid po kateremkoli izmed zahtevkov od 10 do 12, značilen po tem, da je pripravljen s pomočjo kemijske sinteze.A polypeptide according to any one of claims 10 to 12, characterized in that it is prepared by chemical synthesis.15. Polipeptid po kateremkoli izmed zahtevkov od 10 do 14, značilen po tem, da se omenjeno zaporedje sosednjih amino kislin nahaja na sliki 14.A polypeptide according to any one of claims 10 to 14, characterized in that said sequence of adjacent amino acids is found in Figure 14.16. Polipeptid po kateremkoli izmed zahtevkov od 10 do 14, značilen po tem, da se omenjeno zaporedje sosednjih amino kislin nahaja na sliki 47.A polypeptide according to any one of claims 10 to 14, characterized in that said sequence of adjacent amino acids is found in Figure 47.17. Polipeptid po kateremkoli izmed zahtevkov od 10 do 14, značilen po tem, da je omenjeno zaporedje sosednjih amino kislin kodirano znotraj Iambda-gt11 cDNA knjižnice, ki je deponirana pri American Type Culture Collection (ATTC - Ameriška zbirka tipskih kultur; Op. prev.) pod številko 40394.A polypeptide according to any one of claims 10 to 14, characterized in that said sequence of adjacent amino acids is encoded within the Iambda-gt11 cDNA library deposited with the American Type Culture Collection (ATTC); .) under number 40394.18. Prečiščen polipeptid po kateremkoli zahtevku od 10 do 17, značilen po tem, da je polipeptid fiksiran na trdno fazo.Purified polypeptide according to any one of claims 10 to 17, characterized in that the polypeptide is fixed to the solid phase.19. Oprema za analizo bioloških vzorcev na prisotnost antiteles usmerjenih proti HCV antigenu, značilna po tem, da obsega polipeptid po kateremkoli zahtevku od 10 do 18 v primerni posodi.Equipment for analyzing biological samples for the presence of antibodies directed against HCV antigen, characterized in that it comprises a polypeptide according to any one of claims 10 to 18 in a suitable container.20. Imuno preiskava za detekcijo antiteles usmerjenih proti virusu hepatitisa C (anti-HCV antitelo), značilna po tem, da obsega: a) zagotovitev polipeptida, ki vsebuje antigensko determinanto, ki je sposobna vezave s pomočjo omenjenega anti-HCV antitelesa, kjer omenjena antigenska determinanta vsebuje zaporedje sosednjih amino kislin, ki je kodirano s HCV genomom in kjer je HCV značilen po tem, da: (i) ima pozitivne RNA genome; (ii) omenjeni genom vsebuje odprto mesto čitanja (ORF - open reading frame), ki kodira poliprotein; in 3 (iii) omenjeni poliprotein vsebuje zaporedje amino kislin, ki najmanj 40 % ustreza zaporedju amino kislin 859 na sliki 14; b) inkubacijo biološkega vzorca z omenjenim polipeptidom po kateremkoli zahtevku od 10 do 18 pri pogojih, ki dovoljujejo nastanek kompleksa antitelo-antigen; in c) ugotavljanje, ali je nastal kompleks antitelo-antigen, ki vsebuje omenjeni polipeptid.20. An immunoassay for the detection of antibodies directed against hepatitis C virus (anti-HCV antibody), characterized in that it comprises: a) providing a polypeptide containing an antigenic determinant capable of binding by said anti-HCV antibody, wherein said the antigenic determinant comprises a sequence of adjacent amino acids encoded by the HCV genome and wherein HCV is characterized in that: (i) it has positive RNA genomes; (ii) said genome contains an open reading frame (ORF) encoding a polyprotein; and 3 (iii) said polyprotein contains an amino acid sequence that corresponds to at least 40% of amino acid sequence 859 in Figure 14; b) incubating a biological sample with said polypeptide according to any one of claims 10 to 18 under conditions that allow the formation of an antibody-antigen complex; and c) determining whether an antibody-antigen complex containing said polypeptide has formed.21. Sestavek za cepljenje, značilen po tem, da vsebuje prečiščeni polipeptid po kateremkoli zahtevku od 10 do 17, pomešan s farmacevtsko sprejemljivim ekscipientom.A vaccine composition comprising a purified polypeptide according to any one of claims 10 to 17 in admixture with a pharmaceutically acceptable excipient.22. Sestavek za cepljenje po zahtevku 21, značilen po tem, da je omenjena sosednja sekvenca iz strukture HCV proteina.The vaccine composition according to claim 21, characterized in that said adjacent sequence is from the structure of the HCV protein.23. Polinukleotid v v bistvu izolirani obliki, ki obsega zaporedje sosednjih polinukleotidov, ki se je sposobno selektivno hibridizirati v genom virusa hepatitisa C (HCV) ali v njegov komplement, kjer je HCV značilen po tem, da (i) ima pozitivne RNA genome; (ii) omenjeni genom vsebuje odprto mesto čitanja (ORF - open reading frame), ki kodira poliprotein; in (iii) omenjeni poliprotein vsebuje zaporedje amino kislin, ki najmanj 40 % ustreza zaporedju amino kislin 859 na sliki 14.23. A polynucleotide in a substantially isolated form comprising a sequence of contiguous polynucleotides capable of selectively hybridizing to the hepatitis C virus (HCV) genome or to its complement, wherein HCV is characterized in that (i) it has positive RNA genomes; (ii) said genome contains an open reading frame (ORF) encoding a polyprotein; and (iii) said polyprotein contains an amino acid sequence that corresponds to at least 40% of amino acid sequence 859 in Figure 14.24. Polinukleotid po zahtevku 23, značilen po tem, da omenjeni poliprotein vsebuje zaporedje aminokislin, ki najmanj 60 % ustreza zaporedju amino kislin 859 na sliki 14.The polynucleotide of claim 23, wherein said polyprotein comprises an amino acid sequence that corresponds to at least 60% of the amino acid sequence 859 of Figure 14.25. Polinukleotid po zahtevku 23 ali 24, značilen po tem, da omenjeno sosednje zaporedje obsega vsaj 10 nukleotidov.Polynucleotide according to claim 23 or 24, characterized in that said adjacent sequence comprises at least 10 nucleotides.26. Polinukleotid po zahtevku 25, značilen po tem, da omenjeno sosednje zaporedje obsega vsaj 15 nukleotidov.The polynucleotide of claim 25, wherein said adjacent sequence comprises at least 15 nucleotides.27. Polinukleotid po zahtevku 26, značilen po tem, da omenjeno sosednje zaporedje obsega vsaj 20 nukleotidov.The polynucleotide of claim 26, wherein said adjacent sequence comprises at least 20 nucleotides.28. Polinukleotid po kateremkoli izmed zahtevkov od 23 do 27, značilen po tem, da je DNA polinukleotid.Polynucleotide according to any one of claims 23 to 27, characterized in that the DNA is a polynucleotide.29. Polinukleotid po kateremkoli izmed zahtevkov od 23 do 28, značilen po tem, da je fiksiran na trdno fazo.Polynucleotide according to any one of claims 23 to 28, characterized in that it is fixed to the solid phase.30. Sonda, značilna po tem, da vsebuje polinukleotid po kateremkoli zahtevku od 23 do 29, ki nadalje vsebuje zaznavno oznako.A probe comprising a polynucleotide according to any one of claims 23 to 29, further comprising a detectable label.31. Oprema za analizo vzorcev na prisotnost polinukleotidov izvedenih iz HCV, značilna po tem, da obsega polinukleotidno sondo po kateremkoli zahtevku od 23 do 30 v primerni posodi.Equipment for analyzing samples for the presence of polynucleotides derived from HCV, characterized in that it comprises a polynucleotide probe according to any one of claims 23 to 30 in a suitable container.32. Postopek preiskave vzorca na prisotnost ali odsotnost HCV polinukleotidov, značilen po tem, da obsega: a) kontaktiranje vzorca s sondo, ki vsebuje polinukleotid po kateremkoli izmed zahtevkov od 23 do 30 pri pogojih, ki dovoljujejo selektivno hibridizacijo omenjene sonde v HCV polinukleotid ali v njegov komplement v vzorcu; b) ugotavljanje, ali so nastali polinukleotidni dupleksi, ki vsebujejo omenjeno sondo.A method of examining a sample for the presence or absence of HCV polynucleotides, characterized in that it comprises: a) contacting the sample with a probe containing a polynucleotide according to any one of claims 23 to 30 under conditions allowing selective hybridization of said probe to an HCV polynucleotide; in its complement in the sample; b) determining whether polynucleotide duplexes containing said probe have formed.
SI8812138A1987-11-181988-11-18Nanbv diagnostics and vaccines
SI8812138B
(en)