Peptides derived from viral proteins for use as immunogens and dosage reactants

Abstract

The present invention relates to novel peptides and methods for treatment, diagnosis and prognosis of virus infections including infections with HCV, HIV, HPV, CMV and Influenza. The invention further relates to methods for identifying and providing peptides useful for the treatment and diagnosis.

Claims

1. An isolated monomeric peptide comprising the following structure
Z.sup.1-Z.sup.2-Z.sup.3-Z.sup.4-Z.sup.6-Z.sup.7-Z.sup.9 wherein Z.sup.1, Z.sup.4, and optional Z.sup.7 defines a linear sequence of one, two, or three arginine residues or derivatives thereof optionally followed by a glycine (G); Z.sup.2 defines an optional amino acid selected from 2,3-Diaminopropionic acid (Dpr), Cysteine, Lysine, Aspartic acid, and Glutamic acid, or a derivative thereof; Z.sup.3 selected from the group consisting of GGQLIGGIYLIPG (SEQ ID NO:313), VITYSIFLIVS (SEQ ID NO:314), TANWARVIS (SEQ ID NO:315), GYLPAVGAPI (SEQ ID NO:316), NIVP(Nle)VVTA (SEQ ID NO:317), VTPADLIGA (SEQ ID NO:318), PRPEGYTLFF (SEQ ID NO:319), LPYPRGYTLFV (SEQ ID NO:320), ETILTPRDV (SEQ ID NO:321), SSTSPVYDL (SEQ ID NO:322), TAYER(Nle)CNIL (SEQ ID NO:323), TVIGAS(Nle)IPLL (SEQ ID NO:324), AAFEE(Nle)(Har)ITS (SEQ ID NO:325), GLEPLVIAGILA (SEQ ID NO:326), TAFLVRNVA (SEQ ID NO:327), TPI(Har)QDWGNRAN (SEQ ID NO:328), TPT(Har)NGWDVKLS (SEQ ID NO:329), LECVYCKQQLL (SEQ ID NO:330), GVYDFAFRDLC (SEQ ID NO:331), GVFDYAFRDIN (SEQ ID NO:332), and VDIRTLEDLL (SEQ ID NO:333); wherein Z.sup.6 is selected from the group consisting of EVYDFAFRDLC (SEQ ID NO:334), GFAFRDLCIVY (SEQ ID NO:335), GFAYRDINLAY (SEQ ID NO:336), GTLGIVCPIG (SEQ ID NO:337), GLEPLVIAGILA (SEQ ID NO:338), TPI(Har)QDWENRAN (SEQ ID NO:339), VAFEDL(Har)(Nle)(Nle)SFI (SEQ ID NO:340), RFQTVVQBA (SEQ ID NO:341), GSLVGLLHIVL (SEQ ID NO:342), SIARSVTI(Nle)(Har)ASVVH (SEQ ID NO:343), TPTRQEWDCRIS (SEQ ID NO:344), TPTRQEWDARIS (SEQ ID NO:345), TPI(Har)QEW(Har)SL(Nle)NQEW (SEQ ID NO:346), IGDLIVAQV (SEQ ID NO:347), QYNPVAV(Nle)F (SEQ ID NO:348), GYTLFFTS (SEQ ID NO:349), GYTLFVSD (SEQ ID NO:350), NTL(Nle)TPRDV (SEQ ID NO:351), SSTSPVYNL (SEQ ID NO:352), VITFSIYLIVS (SEQ ID NO:353), GGNVIGGIY(Nle)IPR (SEQ ID NO:354), ANWAKVIL (SEQ ID NO:355), VIRVIAHGLRL (SEQ ID NO:356), and IGDLIVQAV (SEQ ID NO:478); and wherein optional Z.sup.9 is NWAKVI, wherein Nle is Norleucine and Har is homoarginine.

2. An isolated dimeric or multimeric peptide comprising two or more covalently joined monomeric peptides, each monomeric peptide independently comprising the following structure
Z.sup.1-Z.sup.2-Z.sup.3-Z.sup.4-Z.sup.6-Z.sup.7-Z.sup.9 wherein Z.sup.1, Z.sup.4, and optional Z.sup.7 defines a linear sequence of one, two, or three arginine residues or derivatives thereof optionally followed by a glycine (G); Z.sup.2 defines an optional amino acid selected from 2,3-Diaminopropionic acid (Dpr), Cysteine, Lysine, Aspartic acid and Glutamic acid, or a derivative thereof; wherein Z.sup.3 is selected from the group consisting of GGQLIGGIYLIPG (SEQ ID NO:313), VITYSIFLIVS (SEQ ID NO:314), TANWARVIS (SEQ ID NO:315), GYLPAVGAPI (SEQ ID NO:316), NIVP(Nle)VVTA (SEQ ID NO:317), VTPADLIGA (SEQ ID NO:318), PRPEGYTLFF (SEQ ID NO:319), LPYPRGYTLFV (SEQ ID NO:320), ETILTPRDV (SEQ ID NO:321), SSTSPVYDL (SEQ ID NO:322), TAYER(Nle)CNIL (SEQ ID NO:323), TVIGAS(Nle)IPLL (SEQ ID NO:324), AAFEE(Nle)(Har)ITS (SEQ ID NO:325), GLEPLVIAGILA (SEQ ID NO:326), TAFLVRNVA (SEQ ID NO:327), TPI(Har)QDWGNRAN (SEQ ID NO:328), TPT(Har)NGWDVKLS (SEQ ID NO:329), LECVYCKQQLL (SEQ ID NO:330), GVYDFAFRDLC (SEQ ID NO:331), GVFDYAFRDIN (SEQ ID NO:332), and VDIRTLEDLL (SEQ ID NO:333); wherein Z.sup.6 is selected from the group consisting of EVYDFAFRDLC (SEQ ID NO:334), GFAFRDLCIVY (SEQ ID NO:335), GFAYRDINLAY (SEQ ID NO:336), GTLGIVCPIG (SEQ ID NO:337), GLEPLVIAGILA (SEQ ID NO:338), TPI(Har)QDWENRAN (SEQ ID NO:339), VAFEDL(Har)(Nle)(NLE)SFI (SEQ ID NO:340), RFQTVVQBA (SEQ ID NO:341), GSLVGLLHIVL (SEQ ID NO:342), SIARSVTI(Nle)(Har)ASVVH (SEQ ID NO:343), TPTRQEWDCRIS (SEQ ID NO:344), TPTRQEWDARIS (SEQ ID NO:345), TPI(Har)QEW(Har)SL(Nle)NQEW (SEQ ID NO:346), IGDLIVAQV (SEQ ID NO:347), QYNPVAV(Nle)F (SEQ ID NO:348), GYTLFFTS (SEQ ID NO:349), GYTLFVSD (SEQ ID NO:350), NTL(Nle)TPRDV (SEQ ID NO:351), SSTSPVYNL (SEQ ID NO:352), VITFSIYLIVS (SEQ ID NO:353), GGNVIGGIY(Nle)IPR (SEQ ID NO:354), ANWAKVIL (SEQ ID NO:355), VIRVIAHGLRL (SEQ ID NO:356), and IGDLIVQAV (SEQ ID NO:478); and wherein optional Z.sup.9 is NWAKVI, wherein Nle is Norleucine and Har is homoarginine.

3. The isolated dimeric or multimeric peptide according to claim 2, wherein two or more monomeric peptides are different in sequence.

4. A composition comprising two or more compounds selected from a monomeric peptide is as defined in claim 1, and an isolated dimeric or multimeric peptide comprising two or more covalently joined monomeric peptides, each monomeric peptide independently comprising the following structure
Z.sup.1-Z.sup.2-Z.sup.3-Z.sup.4-Z.sup.6-Z.sup.7-Z.sup.9 wherein Z.sup.1, Z.sup.4, and optional Z.sup.7 defines a linear sequence of one, two, or three arginine residues or derivatives thereof optionally followed by a glycine (G); Z.sup.2 defines an optional amino acid selected from 2,3-Diaminopropionic acid (Dpr), Cysteine, Lysine, Aspartic acid and Glutamic acid, or a derivative thereof; wherein Z.sup.3 is selected from the group consisting of GGQLIGGIYLIPG (SEQ ID NO:313), VITYSIFLIVS (SEQ ID NO:314), TANWARVIS (SEQ ID NO:315), GYLPAVGAPI (SEQ ID NO:316), NIVP(Nle)VVTA (SEQ ID NO:317), VTPADLIGA (SEQ ID NO:318), PRPEGYTLFF (SEQ ID NO:319), LPYPRGYTLFV (SEQ ID NO:320), ETILTPRDV (SEQ ID NO:321), SSTSPVYDL (SEQ ID NO:322), TAYER(Nle)CNIL (SEQ ID NO:323), TVIGAS(Nle)IPLL (SEQ ID NO:324), AAFEE(Nle)(Har)ITS (SEQ ID NO:325), GLEPLVIAGILA (SEQ ID NO:326), TAFLVRNVA (SEQ ID NO:327), TPI(Har)QDWGNRAN (SEQ ID NO:328), TPT(Har)NGWDVKLS (SEQ ID NO:329), LECVYCKQQLL (SEQ ID NO:330), GVYDFAFRDLC (SEQ ID NO:331), GVFDYAFRDIN (SEQ ID NO:332), and VDIRTLEDLL (SEQ ID NO:333); wherein Z.sup.6 is selected from the group consisting of EVYDFAFRDLC (SEQ ID NO:334), GFAFRDLCIVY (SEQ ID NO:335), GFAYRDINLAY (SEQ ID NO:336), GTLGIVCPIG (SEQ ID NO:337), GLEPLVIAGILA (SEQ ID NO:338), TPI(Har)QDWENRAN (SEQ ID NO:339), VAFEDL(Har)(Nle)(Nle)SFI (SEQ ID NO:340), RFQTVVQBA (SEQ ID NO:341), GSLVGLLHIVL (SEQ ID NO:342), SIARSVTI(Nle)(Har)ASVVH (SEQ ID NO:343), TPTRQEWDCRIS (SEQ ID NO:344), TPTRQEWDARIS (SEQ ID NO:345), TPI(Har)QEW(Har)SL(Nle)NQEW (SEQ ID NO:346), IGDLIVAQV (SEQ ID NO:347), QYNPVAV(Nle)F (SEQ ID NO:348), GYTLFFTS (SEQ ID NO:349), GYTLFVSD (SEQ ID NO:350), NTL(Nle)TPRVD (SEQ ID NO:351), SSTSPVYNL (SEQ ID NO:352), VITFSIYLIVS (SEQ ID NO:353), GGNVIGGIY(Nle)IPR (SEQ ID NO:354), ANWAKVIL (SEQ ID NO:355), VIRVIAHGLRL (SEQ ID NO:356), and IGDLIVQAV (SEQ ID NO:478); and wherein optional Z.sup.9 is NWAKVI, wherein Nle is Norleucine and Har is homoarginine.

5. An immunogenic composition comprising at least one monomeric peptide according to claim 1, and an isolated dimeric or multimeric peptide comprising two or more covalently joined monomeric peptides, each monomeric peptide independently comprising the following structure
Z.sup.1-Z.sup.2-Z.sup.3-Z.sup.4-Z.sup.6-Z.sup.7-Z.sup.9 wherein Z.sup.1, Z.sup.4, and optional Z.sup.7 defines a linear sequence of one, two, or three arginine residues or derivatives thereof optionally followed by a glycine (G); Z.sup.2 defines an optional amino acid selected from 2,3-Diaminopropionic acid (Dpr), Cysteine, Lysine, Aspartic acid and Glutamic acid, or a derivative thereof; wherein Z.sup.3 is selected from the group consisting of GGQLIGGIYLIPG (SEQ ID NO:313), VITYSIFLIVS (SEQ ID NO:314), TANWARVIS (SEQ ID NO:315), GYLPAVGAPI (SEQ ID NO:316), NIVP(Nle)VVTA (SEQ ID NO:317), VTPADLIGA (SEQ ID NO:318), PRPEGYTLFF (SEQ ID NO:319), LPYPRGYTLFV (SEQ ID NO:320), ETILTPRDV (SEQ ID NO:321), SSTSPVYDL (SEQ ID NO:322), TAYER(Nle)CNIL (SEQ ID NO:323), TVIGAS(Nle)IPLL (SEQ ID NO:324), AAFEE(Nle)(Har)ITS (SEQ ID NO:325), GLEPLVIAGILA (SEQ ID NO:326), TAFLVRNVA (SEQ ID NO:327), TPI(Har)QDWGNRAN (SEQ ID NO:328), TPT(Har)NGWDVKLS (SEQ ID NO:329), LECVYCKQQLL (SEQ ID NO:330), GVYDFAFRDLC (SEQ ID NO:331), GVFDYAFRDIN (SEQ ID NO:332), and VDIRTLEDLL (SEQ ID NO:333); wherein Z.sup.6 is selected from the group consisting of EVYDFAFRDLC (SEQ ID NO:334), GFAFRDLCIVY (SEQ ID NO:335), GFAYRDINLAY (SEQ ID NO:336), GTLGIVCPIG (SEQ ID NO:337), GLEPLVIAGILA (SEQ ID NO:338), TPI(Har)QDWENRAN (SEQ ID NO:339), VAFEDL(Har)(Nle)(Nle)SFI (SEQ ID NO:340), RFQTVVQBA (SEQ ID NO:341), GSLVGLLHIVL (SEQ ID NO:342), SIARSVTI(Nle)(Har)ASVVH (SEQ ID NO:343), TPTRQEWDCRIS (SEQ ID NO:344), TPTRQEWDARIS (SEQ ID NO:345), TPI(Har)QEW(Har)SL(Nle)NQEW (SEQ ID NO:346), IGDLIVAQV (SEQ ID NO:347), QYNPVAV(Nle)F (SEQ ID NO:348), GYTLFFTS (SEQ ID NO:349), GYTLFVSD (SEQ ID NO:350), NTL(Nle)TPRDV (SEQ ID NO:351), SSTSPVYNL (SEQ ID NO:352), VITFSIYLIVS (SEQ ID NO:353), GGNVIGGIY(Nle)IPR (SEQ ID NO:354), ANWAKVIL (SEQ ID NO:355), VIRVIAHGLRL (SEQ ID NO:356), and IGDLIVQAV (SEQ ID NO:478); and wherein optional Z.sup.9 is NWAKVI, wherein Nle is Norleucine and Har is homoarginine.

6. The immunogenic composition according to claim 5 in the form of a vaccine composition.

7. A method for inducing an immune response in a subject against an antigen which comprises administration of at least one monomeric peptide according to claim 1, an isolated dimeric or multimeric peptide comprising two or more covalently joined monomeric peptides, each monomeric peptide independently comprising the following structure
Z.sup.1-Z.sup.2-Z.sup.3-Z.sup.4-Z.sup.6-Z.sup.7-Z.sup.9 wherein Z.sup.1, Z.sup.4, and optional Z.sup.7 defines a linear sequence of one, two, or three arginine residues or derivatives thereof optionally followed by a glycine (G); Z.sup.2 defines an optional amino acid selected from 2,3-Diaminopropionic acid (Dpr), Cysteine, Lysine, Aspartic acid, and Glutamic acid, or a derivative thereof; wherein Z.sup.3 is selected from the group consisting of GGQLIGGIYLIPG (SEQ ID NO:313), VITYSIFLIVS (SEQ ID NO:314), TANWARVIS (SEQ ID NO:315), GYLPAVGAPI (SEQ ID NO:316), NIVP(Nle)VVTA (SEQ ID NO:317), VTPADLIGA (SEQ ID NO:318), PRPEGYTLFF (SEQ ID NO:319), LPYPRGYTLFV (SEQ ID NO:320), ETILTPRDV (SEQ ID NO:321), SSTSPVYDL (SEQ ID NO:322), TAYER(Nle)CNIL (SEQ ID NO:323), TVIGAS(Nle)IPLL (SEQ ID NO:324), AAFEE(Nle)(Har)ITS (SEQ ID NO:325), GLEPLVIAGILA (SEQ ID NO:326), TAFLVRNVA (SEQ ID NO:327), TPI(Har)QDWGNRAN (SEQ ID NO:328), TPT(Har)NGWDVKLS (SEQ ID NO:329), LECVYCKQQLL (SEQ ID NO:330), GVYDFAFRDLC (SEQ ID NO:331), GVFDYAFRDIN (SEQ ID NO:332), and VDIRTLEDLL (SEQ ID NO:333); wherein Z.sup.6 is selected from the group consisting of EVYDFAFRDLC (SEQ ID NO:334), GFAFRDLCIVY (SEQ ID NO:335), GFAYRDINLAY (SEQ ID NO:336), GTLGIVCPIG (SEQ ID NO:337), GLEPLVIAGILA (SEQ ID NO:338), TPI(Har)QDWENRAN (SEQ ID NO:339), VAFEDL(Har)(Nle)(Nle)SFI (SEQ ID NO:340), RFQTVVQBA (SEQ ID NO:341), GSLVGLLHIVL (SEQ ID NO:342), SIARSVTI(Nle)(Har)ASVVH (SEQ ID NO:343), TPTRQEWDCRIS (SEQ ID NO:344), TPTRQEWDARIS (SEQ ID NO:345), TPI(Har)QEW(Har)SL(Nle)NQEW (SEQ ID NO:346), IGDLIVAQV (SEQ ID NO:347), QYNPVAV(Nle)F (SEQ ID NO:348), GYTLFFTS (SEQ ID NO:349), GYTLFVSD (SEQ ID NO:350), NTL(Nle)TPRDV (SEQ ID NO:351), SSTSPVYNL (SEQ ID NO:352), VITFSIYLIVS (SEQ ID NO:353), GGNVIGGIY(Nle)IPR (SEQ ID NO:354), ANWAKVIL (SEQ ID NO:355), VIRVIAHGLRL (SEQ ID NO:356), and IGDLIVQAV (SEQ ID NO:478); and wherein optional Z.sup.9 is NWAKVI, wherein Nle is Norleucine and Har is homoarginine, a peptide composition comprising two or more compounds selected from a monomeric peptide is as defined in claim 1, and an isolated dimeric or multimericpeptide comprising two or more monomeric peptides, each monomeric peptide independently comprising the following structure
Z.sup.1-Z.sup.2-Z.sup.3-Z.sup.4-Z.sup.6-Z.sup.7-Z.sup.9 wherein Z.sup.1, Z.sup.4, and optional Z.sup.7 defines a linear sequence of one, two, or three arginine residues or derivatives thereof optionally followed by a glycine (G); Z.sup.2 defines an optional amino acid selected from 2,3-Diaminopropionic acid (Dpr), Cysteine, Lysine, Aspartic acid, and Glutamic acid, or a derivative thereof; wherein Z.sup.3 is selected from the group consisting of GGQLIGGIYLIPG (SEQ ID NO:313), VITYSIFLIVS (SEQ ID NO:314), TANWARVIS (SEQ ID NO:315), GYLPAVGAPI (SEQ ID NO:316), NIVP(Nle)VVTA (SEQ ID NO:317), VTPADLIGA (SEQ ID NO:318), PRPEGYTLFF (SEQ ID NO:319), LPYPRGYTLFV (SEQ ID NO:320), ETILTPRDV (SEQ ID NO:321), SSTSPVYDL (SEQ ID NO:322), TAYER(Nle)CNIL (SEQ ID NO:323), TVIGAS(Nle)IPLL (SEQ ID NO:324), AAFEE(Nle)(Har)ITS (SEQ ID NO:325), GLEPLVIAGILA (SEQ ID NO:326), TAFLVRNVA (SEQ ID NO:327), TPI(Har)QDWGNRAN (SEQ ID NO:328), TPT(Har)NGWDVKLS (SEQ ID NO:329), LECVYCKQQLL (SEQ ID NO:330), GVYDFAFRDLC (SEQ ID NO:331), GVFDYAFRDIN (SEQ ID NO:332), and VDIRTLEDLL (SEQ ID NO:333); wherein Z.sup.6 is selected from the group consisting of EVYDFAFRDLC (SEQ ID NO:334), GFAFRDLCIVY (SEQ ID NO:335), GFAYRDINLAY (SEQ ID NO:336), GTLGIVCPIG (SEQ ID NO:337), GLEPLVIAGILA (SEQ ID NO:338), TPI(Har)QDWENRAN (SEQ ID NO:339), VAFEDL(Har)(Nle)(Nle)SFI (SEQ ID NO:340), RFQTVVQBA (SEQ ID NO:341), GSLVGLLHIVL (SEQ ID NO:342), SIARSVTI(Nle)(Har)ASVVH (SEQ ID NO:343), TPTRQEWDCRIS (SEQ ID NO:344), TPTRQEWDARIS (SEQ ID NO:345), TPI(Har)QEW(Har)SL(Nle)NQEW (SEQ ID NO:346), IGDLIVAQV (SEQ ID NO:347), QYNPVAV(Nle)F (SEQ ID NO:348), GYTLFFTS (SEQ ID NO:349), GYTLFVSD (SEQ ID NO:350), NTL(Nle)TPRDV (SEQ ID NO:351), SSTSPVYNL (SEQ ID NO:352), VITFSIYLIVS (SEQ ID NO:353), GGNVIGGIY(Nle)IPR (SEQ ID NO:354), ANWAKVIL (SEQ ID NO:355), VIRVIAHGLRL (SEQ ID NO:356), and IGDLIVQAV (SEQ ID NO:478); and wherein optional Z.sup.9 is NWAKVI, wherein Nle is Norleucine and Har is homoarginine.

8. A peptide according to claim 1, a peptide composition comprising two or more compounds selected from a monomeric peptide is as defined in claim 1, and an isolated dimeric or multimeric dimeric peptide comprising two or more covalently joined monomeric peptides, each monomeric peptide independently comprising the following structure
Z.sup.1-Z.sup.2-Z.sup.3-Z.sup.4-Z.sup.6-Z.sup.7-Z.sup.9 wherein Z.sup.1, Z.sup.4, and optional Z.sup.7 defines a linear sequence of one, two, or three arginine residues or derivatives thereof optionally followed by a glycine (G); Z.sup.2 defines an optional amino acid selected from 2,3-Diaminopropionic acid (Dpr), Cysteine, Lysine, Aspartic acid and Glutamic acid, or a derivative thereof; wherein Z.sup.3 is selected from the group consisting of GGQLIGGIYLIPG (SEQ ID NO:313), VITYSIFLIVS (SEQ ID NO:314), TANWARVIS (SEQ ID NO:315), GYLPAVGAPI (SEQ ID NO:316), NIVP(Nle)VVTA (SEQ ID NO:317), VTPADLIGA (SEQ ID NO:318), PRPEGYTLFF (SEQ ID NO:319), LPYPRGYTLFV (SEQ ID NO:320), ETILTPRDV (SEQ ID NO:321), SSTSPVYDL (SEQ ID NO:322), TAYER(Nle)CNIL (SEQ ID NO:323), TVIGAS(Nle)IPLL (SEQ ID NO:324), AAFEE(Nle)(Har)ITS (SEQ ID NO:325), GLEPLVIAGILA (SEQ ID NO:326), TAFLVRNVA (SEQ ID NO:327), TPI(Har)QDWGNRAN (SEQ ID NO:328), TPT(Har)NGWDVKLS (SEQ ID NO:329), LECVYCKQQLL (SEQ ID NO:330), GVYDFAFRDLC (SEQ ID NO:331), GVFDYAFRDIN (SEQ ID NO:332), and VDIRTLEDLL (SEQ ID NO:333); wherein Z.sup.6 is selected from the group consisting of EVYDFAFRDLC (SEQ ID NO:334), GFAFRDLCIVY (SEQ ID NO:335), GFAYRDINLAY (SEQ ID NO:336), GTLGIVCPIG (SEQ ID NO:337), GLEPLVIAGILA (SEQ ID NO:338), TPI(Har)QDWENRAN (SEQ ID NO:339), VAFEDL(Har)(Nle)SFI (SEQ ID NO:340), RFQTVVQBA (SEQ ID NO:341), GSLVGLLHIVL (SEQ ID NO:342), SIARSVTI(Nle)(Har)ASVVH (SEQ ID NO:343), TPTRQEWDCRIS (SEQ ID NO:344), TPTRQEWDARIS (SEQ ID NO:345), TPI(Har)QEW(Har)SL(Nle)NQEW (SEQ ID NO:346), IGDLIVAQV (SEQ ID NO:347), QYNPVAV(Nle)F (SEQ ID NO:348), GYTLFFTS (SEQ ID NO:349), GYTLFVSD (SEQ ID NO:350), NTL(Nle)TPRDV (SEQ ID NO:351), SSTSPVYNL (SEQ ID NO:352), VITFSIYLIVS (SEQ ID NO:353), GGNVIGGIY(Nle)IPR (SEQ ID NO:354), ANWAKVIL (SEQ ID NO:355), VIRVIAHGLRL (SEQ ID NO:356), and IGDLIVQAV (SEQ ID NO:478); and wherein optional Z.sup.9 is NWAKVI, for use as a medicament, wherein Nle is Norleucine and Har is homoarginine.

9. A peptide according to claim 1, a peptide composition comprising two or more compounds selected from a monomeric peptide is as defined in claim 1, and an isolated dimeric or multimeric peptide comprising two or more covalently joined monomeric peptides, each monomeric peptide independently comprising the following structure
Z.sup.1-Z.sup.2-Z.sup.3-Z.sup.4-Z.sup.6-Z.sup.7-Z.sup.9 wherein Z.sup.1, Z.sup.4, and optional Z.sup.7 defines a linear sequence of one, two, or three arginine residues or derivatives thereof optionally followed by a glycine (G); Z.sup.2 defines an optional amino acid selected from 2,3-Diaminopropionic acid (Dpr), Cysteine, Lysine, Aspartic acid and Glutamic acid, or a derivative thereof; wherein Z.sup.3 is selected from the group consisting of GGQLIGGIYLIPG (SEQ ID NO:313), VITYSIFLIVS (SEQ ID NO:314), TANWARVIS (SEQ ID NO:315), GYLPAVGAPI (SEQ ID NO:316), NIVP(Nle)VVTA (SEQ ID NO:317), VTPADLIGA (SEQ ID NO:318), PRPEGYTLFF (SEQ ID NO:319), LPYPRGYTLFV (SEQ ID NO:320), ETILTPRDV (SEQ ID NO:321), SSTSPVYDL (SEQ ID NO:322), TAYER(Nle)CNIL (SEQ ID NO:323), TVIGA(Nle)IPLL (SEQ ID NO:324), AAFEE(Nle)(Har)ITS (SEQ ID NO:325), GLEPLVIAGILA (SEQ ID NO:326), TAFLVRNVA (SEQ ID NO:327), TPI(Har)QDWGNRAN (SEQ ID NO:328), TPT(Har)NGWDVKLS (SEQ ID NO:329), LECVYCKQQLL (SEQ ID NO:330), GVYDFAFRDLC (SEQ ID NO:331), GVFDYAFRDIN (SEQ ID NO:332), and VDIRTLEDLL (SEQ ID NO:333); wherein Z.sup.6 is selected from the group consisting of EVYDFAFRDLC (SEQ ID NO:334), GFAFRDLCIVY (SEQ ID NO:335), GFAYRDINLAY (SEQ ID NO:336), GTLGIVCPIG (SEQ ID NO:337), GLEPLVIAGILA (SEQ ID NO:338), TPI(Har)QDWENRAN (SEQ ID NO:339), VAFEDL(Har)(Nle)(Nle)SFI (SEQ ID NO:340), RFQTVVQBA (SEQ ID NO:341), GSLVGLLHIVL (SEQ ID NO:342), SIARSVTI(Nle)(Har)ASVVH (SEQ ID NO:343), TPTRQEWDCRIS (SEQ ID NO:344), TPTRQEWDARIS (SEQ ID NO:345), TPI(Har)QEW(Har)SL(Nle)NQEW (SEQ ID NO:346), IGDLIVAQV (SEQ ID NO:347), QYNPVAV(Nle)F (SEQ ID NO:348), GYTLFFTS (SEQ ID NO:349), GYTLFVSD (SEQ ID NO:350), NTL(Nle)TPRDV (SEQ ID NO:351), SSTSPVYNL (SEQ ID NO:352), VITFSIYLIVS (SEQ ID NO:353), GGNVIGGIY(Nle)IPR (SEQ ID NO:354), ANWAKVIL (SEQ ID NO:355), VIRVIAHGLRL (SEQ ID NO:356), and IGDLIVQAV (SEQ ID NO:478); and wherein optional Z.sup.9 is NWAKVI, for treating the pathological effects of a disease antigen wherein Nle is Norleucine, and Har is homoarginine.

Description

LEGENDS TO THE FIGURES

(1) FIG. 1. Intracellular uptake of influenza scaffold peptides. Median and intequartile range of readouts from buffy coats from ten donors and three concentrations of peptide each, normalized by value for N-biotin for each donor.

(2) FIG. 2. Extracellular uptake of influenza scaffold peptides. Median and intequartile range of readouts from buffy coats from ten donors and three concentrations of peptide each, normalized by value for N-biotin for each donor.

(3) FIG. 3. Intracellular uptake of HCV scaffold peptides. Median and intequartile range of readouts from buffy coats from five donors at four different concentrations of peptide each, normalized by value for N-biotin for each donor.

(4) FIG. 4. Extracellular uptake of HCV scaffold peptides. Median and intequartile range of readouts from buffy coats from five donors at four different concentrations of peptide each, normalized by value for N-biotin for each donor.

(5) FIG. 5. Median loss of weight by treatment group after challenge. The median weight by treatment groups; ISA5: peptides and ISA51, Provax: peptides and Provax, PR8: inactivated influenza A/PR8 (H1N1) virus, Nave: no treatment before challenge. For animals lost to humane endpoints a last observation carried forward method was employed for the weights

DETAILED DISCLOSURE OF THE INVENTION

Definitions

(6) When terms such as one, a or an are used in this disclosure they mean at least one, or one or more unless otherwise indicated. Further, the term comprising is intended to mean including and thus allows for the presence of other constituents, features, conditions, or steps than those explicitly recited.

(7) As used herein a multimeric peptide or oligomeric peptide refers to an assembly of two or more different or identical linear peptide sequences or subunits, preferably interconnected or assembled by one or more chemical bond of a linker. Preferably the peptide sequences are interconnected by one or more, such as one covalent bond, such as an intermolecular disulfide (SS) bond between two Cys residues, a methylated peptide bond between a N--methylated Lys side-chain and the side-chain of an Asp or Glu residue, an oxime bond, a thioether bond, or a non-covalent bond, such as in a -stacking of rings wherein a W residue in Z.sup.2 of the first Z.sup.1-Z.sup.2 peptide repeat is linked to an Y residue in Z.sup.2 of the second Z.sup.1-Z.sup.2 peptide repeat. The term includes a dimeric (or dimer) peptide suitably formed by a chemical linking of two linear peptide sequences. The term multimeric peptide further includes an assembly of 2, 3, 4, 5, 6, 7, 8, 9 or 10 different or identical peptide sequences. In some embodiments, the multimeric peptide is a dimeric peptide.

(8) As used herein a linker refers to any compound suitable for assembly of the two or more different or identical linear peptide sequences or subunits into a multimeric peptide, or to any other therapeutically active compound, such as an immunomodulating compound. The term includes any linker found useful in peptide chemistry. Since the multimeric peptide may be assembled or connected by standard peptide bonds in a linear way, the term linker also includes a peptide spacer, also referred to as a spacer.

(9) In some embodiments, the linker is not a peptide sequence. In some embodiments, the linker is not a branched peptide sequence.

(10) In some embodiments, the linker does not itself contain a peptide sequence derived from or identical to a natural antigen.

(11) In some embodiments, the linker has a molecular weight of less than 10 kDa, such as less than 9 kDa, such as less than 8 kDa, such as less than 7 kDa, such as less than 6 kDa, such as less than 5 kDa, such as less than 4 kDa, such as less than 3 kDa, such as less than 2 kDa, such as less than 1.5 kDa, such as less than 1 kDa, such as less than 0.5 kDa, such as less than 0.2 kDa. In some embodiments, wherein the multimeric peptide is a dimeric peptide, the linker is not linking the two peptide sequences from one terminal cysteine in the first peptide to a second terminal cysteine in the second peptide.

(12) In some embodiments, the linker is not linking the two or more peptide sequences through a terminal cysteine in any one of the peptides.

(13) In some embodiments, the linker is not linking from a cysteine residue.

(14) HIV generally denotes human immunodeficiency virus I.

(15) HIV disease is composed of several stages including the acute HIV infection which often manifests itself as an influenza-like infection and the early and medium stage symptomatic disease, which has several non-characteristic symptoms such as skin rashes, fatigue, night sweats, slight weight loss, mouth ulcers, and fungal skin and nail infections. Most HIV infected will experience mild symptoms such as these before developing more serious illnesses. It is generally believed that it takes five to seven years for the first mild symptoms to appear. As HIV disease progresses, some individuals may become quite ill even if they have not yet been diagnosed with AIDS (see below), the late stage of HIV disease. Typical problems include chronic oral or vaginal thrush (a fungal rash or spots), recurrent herpes blisters on the mouth (cold sores) or genitals, ongoing fevers, persistent diarrhea, and significant weight loss. AIDS is the late stage HIV disease and is a condition which progressively reduces the effectiveness of the immune system and leaves individuals susceptible to opportunistic infections and tumors.

(16) The term cell-penetrating peptide as used herein refers to any peptide with the capability to translocate across the plasma membrane into either cytoplasmic and/or nuclear compartments of eukaryotic and/or prokaryotic cells, such as into cytoplasm, nucleus, lysosome, endoplasmatic reticulum, golgi apparatus, mitocondria and/or chloroplast, seemingly energy-independently. This capability to translocate across the plasma membrane of a cell-penetrating peptide according to the invention may be non-invasive, energy-independent, non-saturable, and/or receptor independent. In one embodiment the term cell-penetrating peptide refers to a peptide, which is demonstrated to translocate across a plasma membrane as determined by the assay in example 5. It is to be understood that a cell-penetrating peptide according to the present invention may be translocated across the membrane with the sequence complete and intact, or alternatively partly degraded, but in a form where the antigens contained within this peptide is able to be presented within the cell to stimulate an immune response. Accordingly, a cell-penetrating peptide according to the present invention is a peptide that may be demonstrated to translocate across a plasma membrane as determined by the assay in example 5 and be demonstrated to stimulate an effective immune response.

(17) The monomeric peptide according to the present invention may be provided in any pharmaceutically acceptable salt, such as in a salt of acetat or HCl.

(18) The term derived from an antigen when in reference to a peptide derived from a source (such as a virus etc.) as used herein is intended to refer to a peptide which has been obtained (e.g., isolated, purified, etc.) from the source. Preferably, the peptide may be genetically engineered and/or chemically synthesized to be essentially identical to the native peptide of the source. The term includes the use of variants of known native peptide sequences, such as peptide sequences, where 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acids of the native peptide sequence have been substituted with any other amino acid, such as conservative substitutions. Alternatively, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acids have been removed or added to the native peptide sequence. Accordingly, in some embodiments, the peptides according to the present invention comprises an amino acid sequence Z.sup.3, and optional Z.sup.6, Z.sup.9 and Z.sup.12, that is defined as a sequence of 8-30 amino acids, such as 8-20 amino acids derived from an antigen, wherein the peptide sequence of the antigen comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 substitutions, additions or deletions relative to the antigen, such as the addition of an arginine in the N- or C-terminal of the amino acid sequence of Z.sup.3, and optional Z.sup.6, Z.sup.9 and Z.sup.12. In some embodiments, the peptides according to the present invention comprises an amino acid sequence Z.sup.3, and optional Z.sup.6, Z.sup.9 and Z.sup.12, that is defined as a sequence of 8-30 amino acids, such as 8-20 amino acids identical in sequence to a native antigen. In some embodiments, the peptides according to the present invention comprises an amino acid sequence Z.sup.3, and optional Z.sup.6, Z.sup.9 and Z.sup.12, that is defined as a sequence of 8-30 amino acids, such as 8-20 amino acids that is not identical in sequence to a native antigen.

(19) It is to be understood that derived from an antigen does not exclude that an amino acid sequence defined by Z.sup.3, and optional Z.sup.6, Z.sup.9 and Z.sup.12 may be derived from more than one antigenic peptide sequence, such as from two or three different proteins or peptide sources or different sequences within the same proteins or peptide of the same virus, any different virus, or any disease antigen. However, in one embodiment Z.sup.3, and optional Z.sup.6, Z.sup.9 and Z.sup.12 are derived from one specific continuous peptide sequence. In one embodiment Z.sup.3, and optional Z.sup.6, Z.sup.9 and Z.sup.12 are derived from two different specific continuous peptide sequences of the same or different protein derived from the same virus, any different virus, or any disease antigen.

(20) The amino acids used in the amino acid sequences according to the invention may be in both L- and/or D-form. It is to be understood that both L- and D-forms may be used for different amino acids within the same peptide sequence. In some embodiments the amino acids within the peptide sequence are in L-form, such as natural amino acids. It is to be understood that any known antigen may be used in the constructs according to the present invention.

(21) In some specific embodiments, the first 1, 2, or 3 amino acids in the N-terminal of the amino acid sequences according to the invention are in the D-form. It is assumed that the N-terminal trimming and thereby degradation of the peptides are somewhat delayed by having amino acids of the D-form in the N-terminal of these peptides according to the present invention. Alternatively and in some embodiments, the first 1, 2, or 3 amino acids in the N-terminal of the amino acid sequences according to the invention are amino acids in beta or gamma forms. Beta amino acids have their amino group bonded to the beta carbon rather than the alpha carbon as in the 20 standard natural amino acids.

(22) Alternatively the first 1, 2, or 3 amino acids in the N-terminal of the amino acid sequences according to the invention may be modified by incorporation of fluorine, or alternatively cyclic amino acids or other suitable non-natural amino acids are used.

(23) A variant or analogue of a peptide refers to a peptide having an amino acid sequence that is substantially identical to a reference peptide, typically a native or parent polypeptide. The peptide variant may possess one or more amino acid substitutions, deletions, and/or insertions at certain positions within the native amino acid sequence.

(24) Conservative amino acid substitutions are those in which an amino acid residue is replaced with an amino acid residue having a side chain with similar physicochemical properties. Families of amino acid residues having similar side chains are known in the art, and include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). A particular form of conservative amino acid substitutions include those with amino acids, which are not among the normal 20 amino acids encoded by the genetic code. Since preferred embodiments of the present invention entail use of synthetic peptides, it is unproblematic to provide such non-naturally occurring amino acid residues in the peptides disclosed herein, and thereby it is possible to exchange the natural saturated carbon chains in the side chains of amino acid residues with shorter or longer saturated carbon chainsfor instance, lysine may be substituted with an amino acid having an the side chain (CH.sub.2).sub.nNH.sub.3, where n is different from 4, and arginine may be substituted with an amino acid having the side chain (CH.sub.2).sub.nNHC(NH.sub.2)NH.sub.2, where n is different from 3, etc. Similarly, the acidic amino acids aspartic acid and glutamic acid may be substituted with amino acid residues having the side chains (CH.sub.2).sub.nCOOH, where n>2.

(25) The term substantially identical in the context of two amino acid sequences means that the sequences, when optimally aligned, such as by the programs GAP or BESTFIT using default gap weights, share at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 95, at least about 98, or at least about 99 percent sequence identity. In one embodiment, residue positions that are not identical differ by conservative amino acid substitutions. Sequence identity is typically measured using sequence analysis software. Protein analysis software matches similar sequences using measures of similarity assigned to various substitutions, deletions and other modifications, including conservative amino acid substitutions. For instance, the publicly available GCG software contains programs such as Gap and BestFit which can be used with default parameters to determine sequence homology or sequence identity between closely related polypeptides, such as homologous polypeptides from different species of organisms or between a wild-type protein and a mutein thereof. See, e.g., GCG Version 6.1. Polypeptide sequences can also be compared using FASTA or ClustalW, applying default or recommended parameters. A program in GCG Version 6.1., FASTA (e.g., FASTA2 and FASTA3) provides alignments and percent sequence identity of the regions of the best overlap between the query and search sequences (Pearson, Methods Enzymol. 1990; 183:63-98; Pearson, Methods Mol. Biol. 2000; 132:185-219). Another preferred algorithm when comparing a sequence to a database containing a large number of sequences from various organisms, or when deducing the is the computer program BLAST, especially blastp, using default parameters. See, e.g., Altschul et al., J. Mol. Biol. 1990; 215:403-410; Altschul et al., Nucleic Acids Res. 1997; 25:3389-402 (1997); each herein incorporated by reference. Corresponding amino acid positions in two substantially identical amino acid sequences are those aligned by any of the protein analysis software mentioned herein, typically using default parameters.

(26) An isolated molecule is a molecule that is the predominant species in the composition wherein it is found with respect to the class of molecules to which it belongs (i.e., it makes up at least about 50% of the type of molecule in the composition and typically will make up at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or more of the species of molecule, e.g., peptide, in the composition). Commonly, a composition of a peptide molecule will exhibit 98%-99% homogeneity for peptide molecules in the context of all present peptide species in the composition or at least with respect to substantially active peptide species in the context of proposed use.

(27) The term linear sequence as used herein refers to the specific sequence of amino acids connected by standard peptide bonds in standard N- to C-terminal direction. The peptide may contain only peptide bonds. However the term does not exclude that an amino acid within a sequence, such as within Z.sup.3, may be connected, such as through the side chains, with another amino acid at a distant location within the peptide sequence, such as a distant location within Z.sup.3.

(28) In the context of the present invention, treatment or treating refers to preventing, alleviating, managing, curing or reducing one or more symptoms or clinically relevant manifestations of a disease or disorder, unless contradicted by context. For example, treatment of a patient in whom no symptoms or clinically relevant manifestations of a disease or disorder have been identified is preventive or prophylactic therapy, whereas treatment of a patient in whom symptoms or clinically relevant manifestations of a disease or disorder have been identified generally does not constitute preventive or prophylactic therapy.

(29) The term antigen denotes a substance of matter which is recognized by the immune system's specifically recognizing components (antibodies, T-cells).

(30) The term immunogen is in the present context intended to denote a substance of matter, which is capable of inducing an adaptive immune response in an individual, where said adaptive immune response targets the immunogen. In relation to the present invention, an immunogen will induce a humoral and/or cell-mediated immune response. In other words, an immunogen is an antigen, which is capable of inducing immunity.

(31) The terms epitope, antigenic determinant and antigenic site are used interchangeably herein and denotes the region in an antigen or immunogen which is recognized by antibodies (in the case of antibody binding epitopes, also known as B-cell epitopes) or by T-cell receptors when the epitope is complexed to a Major histocompatibility complex (MHC) molecule (in the case of T-cell receptor binding epitopes, i.e. T-cell epitopes).

(32) B cell antigen means any antigen that naturally is or could be engineered to be recognized by a B cell, and that triggers an immune response in a B cell (e.g., an antigen that is specifically recognized by a B cell receptor on a B cell).

(33) The term immunogenically effective amount has its usual meaning in the art, i.e. an amount of an immunogen, which is capable of inducing an immune response, which significantly engages pathogenic agents, which share immunological features with the immunogen.

(34) The term vaccine is used for a composition comprising an immunogen and which is capable of inducing an immune response which is either capable of reducing the risk of developing a pathological condition or capable of inducing a therapeutically effective immune response which may aid in the cure of (or at least alleviate the symptoms of) a pathological condition.

(35) The term pharmaceutically acceptable has its usual meaning in the art, i.e. it is used for a substance that can be accepted as part of a medicament for human use when treating the disease in question and thus the term effectively excludes the use of highly toxic substances that would worsen rather than improve the treated subject's condition.

(36) A T helper lymphocyte epitope (a T.sub.H epitope) is peptide, which binds an MHC Class II molecule and can be presented on the surface of an antigen presenting cell (APC) bound to the MHC Class II molecule. An immunological carrier is generally a substance of matter which includes one or many T.sub.H epitopes, and which increase the immune response against an antigen to which it is coupled by ensuring that T-helper lymphocytes are activated and proliferate. Examples of known immunological carriers are the tetanus and diphtheria toxoids and keyhole limpet hemocyanin (KLH).

(37) In the scaffold design according to the present invention, Z.sup.3, and optional Z.sup.6, Z.sup.9 and Z.sup.12 may define a sequence of amino acids, such as 8-30 amino acids, such as 8-20 amino acids derived from the antigen. This sequence of amino acids derived from an antigen may herein be referred to as an epitope.

(38) The peptides according to the present invention may be a helper T lymphocyte (HTL) inducing peptide comprising HTL epitopes. A HTL inducing peptide is a HLA Class II binding peptide that is capable of inducing a HTL response. Also the peptides according to the present invention may in other embodiments be CTL inducing peptides comprising CTL epitopes in addition to or as an alternative to being a HTL inducing peptide. A CTL inducing peptide is a HLA Class I binding peptide that is capable of inducing a CTL response.

(39) In some embodiments the epitopes used in the scaffold according to the present invention are CTL epitopes. A CTL inducing peptide is a HLA Class I binding peptide that is capable of inducing a CTL response. In other embodiments the epitopes used in the scaffold design according to the present invention are HTL inducing peptides. A HTL inducing peptide is a HLA Class II binding peptide that is capable of inducing a HTL response.

(40) In other alternative embodiments, tryptophan or tryptophan derivatives are used in the sequence defined by Z.sup.2, Z.sup.5, Z.sup.8 and Z.sup.11. Any suitable tryptophan derivatives may be used. As used herein tryptophan derivatives means an unnatural modified tryptophan amino acid residue including those disclosed in U.S. Pat. No. 7,232,803, such as tri tert.-butyltryptophan, di-tert-butyl tryptophan, 7-benzyloxytryptophan, homotryptophan, 5-aminoethyltryptophan (available as side chain Boc and N-alpha FMOC derivative from RSP Amino Acids Analogues Inc, Boston, Mass., USA), N-Acetylhomotryptophan (Toronto Research), 7-Benzyloxytryptophan (Toronto Research), Homotryptophan (Toronto Research), and tryptophan residues which have been substituted at the 1-, 2-, 5- and/or 7-position of the indole ring, positions 1- or 2- being preferred e.g. 5 hydroxy tryptophan.

(41) The term amino acid derivative, sometimes used in the context of a derivative thereof referring to a specific amino acid, means an amino acid compound, wherein one or more chemical groups has been modified, added or removed as compared to the amino acid to which the amino acid compound is a derivative of, while still having an amine group and a carboxylic acid group, as well as a side chain of an amino acid and still being able to form peptide bonds. In some embodiments an amino acid derivative is a standard amino acid that has only been modified in the side chain of the amino acid. In some embodiments an amino acid derivative is a non-natural amino acid such as Dpr. In some embodiments an amino acid is a modified moiety which is incorporated into the chemically synthesized peptide or polypeptide and that comprises an activatable group that is linkable, after activation, to another peptide, such as Dpr(Ser), Lys(Ser), or Ornithine(Ser).

(42) The term basic amino acid as used herein refers to any amino acid including both natural and non-natural amino acids that has an isoelectric point above 6.3 (such as above 7.4) as measured according to Kice & Marvell Modern Principles of organic Chemistry (Macmillan, 1974) or Matthews and van Holde Biochemistry Cummings Publishing Company, 1996. Included within this definition are Arginine, Lysine, Homoarginine (Har), and Histidine as well as derivatives thereof. Suitable non-natural basic amino acids are e.g. as described in U.S. Pat. No. 6,858,396. Suitable positively charged amino acids includes non-natural alpha amino acids available from Bachem AG and includes alpha-amino-glycine, alpha,gamma-diaminobutyric acid, ornithine, alpha, beta-diaminoproprionic acid, alpha-difluoromethyl-ornithine, 4-amino-piperidine-4-carboxylic acid, 2,6-diamino-4-hexynoic acid, beta-(1-piperazinyl)-alanine, 4,5-dehydro-lysine, delta-hydroxy-lysine, omega-hydroxy-norarginine, homoarginine, omega-amino-arginine, omega-methyl-arginine, alpha-methyl-histidine, 2,5-diiodo-histidine, 1-methyl-histidine, 3-methyl-histidine, beta-(2-pyridyl)-alanine, beta-(3-pyridyl)-alanine, beta-(2-quinolyl)-alanine, 3-amino-tyrosine, 4-amino-phenylalanine, and spinacine. Furthermore, any mono or dicarboxylic amino acid is a suitable positively charged amino acid.

(43) The term neutral amino acid as used herein refers to an amino acid that has an isoelectric point above between 4.8 and 6.3 as measured according to Kice & Marvell Modern Principles of organic Chemistry (Macmillan, 1974). The term acidic amino acid as used herein refers to an amino acid that has an isoelectric point below 4.8 as measured according to Kice & Marvell Modern Principles of organic Chemistry (Macmillan, 1974).

(44) Unless otherwise indicated amino acids are abbreviated and mentioned by their standard nomenclature known to the person skilled in the art, such as with reference to nomenclature and symbolism for amino acids and peptides by the international union of pure and applied chemistry (IUPAC) (www.iupac.org).

(45) The term antibody response refers to the production of antibodies (e.g., IgM, IgA, IgG) which bind to an antigen of interest, this response is measured for instance by assaying sera by antigen ELISA.

(46) The term adjuvant as used herein refers to any compound which, when delivered together or simultaneously with an antigen, non-specifically enhances the immune response to that antigen. Exemplary adjuvants include but are not limited to oil in water and water in oil adjuvants, aluminum-based adjuvants (e.g., AIOH, AIPO4, etc), and Montanide ISA 720.

(47) The terms patient and subject refer to a mammal that may be treated using the methods of the present invention.

(48) As used herein, the term immune response refers to the reactivity of an organism's immune system in response to an antigen. In vertebrates, this may involve antibody production, induction of cell-mediated immunity, and/or complement activation (e.g., phenomena associated with the vertebrate immune system's prevention and resolution of infection by microorganisms). In preferred embodiments, the term immune response encompasses but is not limited to one or more of a lymphocyte proliferative response, a cytokine response, and an antibody response.

(49) The term net charge as used herein with reference to a peptide sequence refers to the total electric charge of the peptide sequence represented by the sum of charges of each individual amino acid in the peptide sequence, wherein each basic amino acid are given a charge of +1, each acidic amino acid a charge of 1, and each neutral amino acid a charge of 0. Accordingly, the net charge will depend on the number and identities of charged amino acids.

(50) Table 1Specific peptides not part of the present invention

(51) Table 1 and 2 represent peptides not part of the present invention comprising the structure
X.sup.1-X.sup.2-X.sup.3-X.sup.4-X.sup.5(formula II),
wherein X.sup.1 and X.sup.3 independently defines a linear sequence of any 1, 2, 3 or 4 amino acid independently selected from any basic amino acid, citrulline, tryptophan, or a derivative thereof; X.sup.2 defines a linear sequence of 8-30 amino acids derived from an antigen; X.sup.4 defines a linear sequence of 8-30 amino acids derived from said antigen, said sequence X.sup.4 being different from X.sup.2; and wherein X.sup.5 is any one optional amino acid selected from a basic amino acid, citrulline, tryptophan, or a derivative thereof. Citrulline is in this document referred to with the one-letter symbol B.

(52) TABLE-US-00001 TABLE1 Placement with referenceto positionsin SEQIDNO:3; SEQIDNO:6; SEQIDNO:7, SEQID NO:11,SEQ IDNO:12, SEQID NO:46,and SEQID NO:126. Reference Modified X2- x4- Antigen ID x1 x2 x3 x4 x5 (m) seq seq P- R QIKIWFQN RR MKWKK biotin SEQIDNO:483 SEQIDNO:484 N- PVVHLTL R QAGDDFSR Biotin SEQIDNO:485 SEQIDNO:486 HCV SP_2 RR GYIPLVGAPLG BGR VARALAHGVRV 135- 147- 145 157 HCV SP_3 R GYIPLVGAPLG RR VARALAHGVRV 135- 147- 145 157 HCV SP_4 R GYIPLVGAPLG RRR VARALAHGVRV R 135- 147- 145 157 HCV SP_5 RR GYIPLVGAPLG RR VARALAHGVRV 135- 147- 145 157 HCV SP_6 RR GYIPLVGAPLG RRR VARALAHGVRV 135- 147- 145 157 HCV SP_7 BR GYIPLVGAPLG RR VARALAHGVRV 135- 147- 145 157 HCV SP_8 RRR GYIPLVGAPLG BR VARALAHGVRV 135- 147- 145 157 HCV SP_9 R GYIPLVGAPLG KKK VARALAHGVRV 135- 147- 145 157 HCV SP_10 R GYIPLVGAPLG RRR VARALAHGVRV 135- 147- 145 157 HCV SP_11 KK GYIPLVGAPLG KK VARALAHGVRV 135- 147- 145 157 HCV SP_12 W GYIPLVGAPLG RR VARALAHGVRV 135- 147- 145 157 HCV SP_13 WW GYIPLVGAPLG RR VARALAHGVRV 135- 147- 145 157 HCV SP_14 EE GYIPLVGAPLG EE VARALAHGVRV 135- 147- 145 157 HCV SP_15 GG GYIPLVGAPLG GG VARALAHGVRV 135- 147- 145 157 HCV SP_16 EE GYIPLVGAPLG RR VARALAHGVRV 135- 147- 145 157 HCV SP_17 RR GYIPLVGAPLG LRR VARALAHGVRV 135- 147- 145 157 HCV SP21: WW GYIPLVGAPLG RR VARALAHGVRV 135- 147- 145 157 HCV SP22: WW GYIPLVGAPLG RRR VARALAHGVRV 135- 147- 145 157 HCV SP23: WW GYIPLVGAPLG R VARALAHGVRV 135- 147- 145 157 HCV SP24: R GYIPLVGAPLG RR VARALAHGVRV 135- 147- 145 157 HCV 51_ RR GYLPAVGAPIG BR VIRVIAHGLRL m 135- 147- BIotin 144 157 HCV 51b_ RR GYIPLVGAPLG BR VARALAHGVRV 135- 147- BIotin 145 157 HCV 51_n GYIPLVGAPLG G VARALAHGVRV 135- 147- 145 157 HCV SP51_1: WW GYLPAVGAPI RR VIRVIAHGLRL m 135- 147- 144 157 HCV SP1_C* GYIPLVGAPLG G VARALAHGVRV 135- 147- 145 157 HCV SP2_c RR GYIPLVGAPLG BGR VARALAHGVRV 135- 147- 145 157 HCV SP3_c R GYIPLVGAPLG RR VARALAHGVRV 135- 147- 145 157 HCV SP4_c R GYIPLVGAPLG RRR VARALAHGVRV 135- 147- 145 157 HCV SP5_c RR GYIPLVGAPLG RR VARALAHGVRV 135- 147- 145 157 HCV SP6_c RR GYIPLVGAPLG RRR VARALAHGVRV 135- 147- 145 157 HCV SP7_c BR GYIPLVGAPLG RR VARALAHGVRV 135- 147- 145 157 HCV SP8_c RRR GYIPLVGAPLG BR VARALAHGVRV 135- 147- 145 157 HCV SP9_c R GYIPLVGAPLG KKK VARALAHGVRV 135- 147- 145 157 HCV SP10_c R GYIPLVGAPLG RRR VARALAHGVRV 135- 147- 145 157 HCV SP11_c KK GYIPLVGAPLG KK VARALAHGVRV 135- 147- 145 157 HCV SP12_c W GYIPLVGAPLG RR VARALAHGVRV 135- 147- 145 157 HCV SP13_c WW GYIPLVGAPLG RR VARALAHGVRV 135- 147- 145 157 HCV SP17_c RR GYIPLVGAPLG LRR VARALAHGVRV 135- 147- 145 157 HCV SP61_2_ RR NYVTGNIPG BR GITFSIFLIVS 163- 171- 171 181 HCV SP61b_ WW NYATGNLPG RR CSFSIFLLAL m 163- 171- 2_ 171 181 HCV SP61_3_ WW NYVTGNIPG BR GITFSIFLIVS 163- 171- 171 181 HCV SP61_4_ WW NYVTGNIPG RR GITFSIFLIVS 163- 171- 171 181 HCV 61b_ RR NYATGNLPG RR GCSFSIFLLAL 163- 171- BIotin 171 181 HCV SP25 RR VTGNIPGSTYS GBR GITFSIYLIVS m 165- 171- 175 181 HCV 42_ RR IRNLGRVIETLTG BR LNleGYIPLIGA m 116- 133- BIotin 128 142 HCV 42b_ RR SRNLGKVIDTLT BR LMGYIPLVGA 116- 133- BIotin C 128 142 HCV 42n- SRNLGKVIDTLT GFAD LMGYIPLVGA 116- 133- BIOTIN C 129 142 HCV SP42_1_ WW IRNLGRVIETLT RR LNleGYIPLIGA m 116- 133- 128 142 HCV SP42b_ WW SRNLGKVIDTLT RR LMGYIPLVGA 116- 133- 1_ C 129 142 HCV BI310- RR GGGQIIGGNYLI PBIGVRATB 26- 42- 11_ P 38 50 Biotin HCV BI310- GGGQIVGGVYLL GPRLGVRATR 26- 42- 11n_ P 38 50 Biotin HCV BI310- RR GGGQIVGGVYLL GPRLGVRATR 26- 42- 11n_sc P 38 50 _Biotin HCV SP11b- WW GGGQIVGGVYLL RR GPRLGVRAT 26- 42- 1- P 38 50 FLU BI100- BR LIFLARSALIV RGSVAHKS 256- 267- 12 266 274 FLU BI100- ED LIFLARSALIL RGSVAHKS 255- 267- 22b 266 274 FLU 120b_ BR LIFLARSALIL BGR SALILRGSVAHK 255- 267- BIotin 266 274 FLU BI100- SAYERMCNIL KGK FQTAAQRAMM 217- 230- 18b 226 239 FLU BI100- SAYERNleVNIL KGK FQTAAQRAVNle 217- 230- 19 226 239 FLU 190_ BR TAYERNleCNIL BRGR FQTVVQBA 217- 230- BIotin 226 237 FLU 190b_ BR IAYERMCNIL LBRGK FQTAAQRA 217- 230- BIotin 226 237 FLU 190n- IAYERMCNIL KGK FQTAAQRA 217- 230- BIOTIN 226 237 FLU BI100- LFFKCIYRLFKHG KR GPSTEGVPESM 46- 62- 24b L 59 72 FLU BI100- BRR LFFKTITRLFBHG RR LLSTEGVPNSNle 46- 62- 26 L 59 72 FLU 260_ BR GLEPLVIAGILA RR GSLVGLLHIVL 23- 30- Biotin 33 40 FLU 260b_ BR GSDPLVVAASIV RR ASIVGILHLIL 23- 30- Biotin 33 40 CMV BI050- R NLVPMVATV RR NLVPMVATV B 485- 485- sc1 493 493 CMV BI050- R NLVPMVATV BRR NLVPMVATV B 485- 485- sc2 493 493 CMV BI050- R NIVPNleVVTA RR NIVPNleVVTA B m 485- 485- sc5 493 493 HIV N10 PEVIPMFSALS EGA TPQDLNTMLN HIV V10 R FIIPXFTALSG GRR ALLYGATPYAIG HIV N13 K ALGPAATL EE MMTACQGVG Neg SP_18 RR GPVVHLTL RR GQAGDDFS c mod Neg SP_19 RR GPVVHLTL RR GQAGDDFS c mod Neg SP_20 RR GPVVHLTL RGR GQAGDDFS c mod HPV RR LECVYCKQQLL RR EVYDFAFRDLC 35- 48- 45 58 HPV RR GVYDFAFRDLC RR GFAFRDLCIVY R 49- 52- 58 61 HPV RR GVFDYAFRDIN RR GFAYRDINLAY R 49- 52- 58 61 CMV RR GATPVDLLGA RR GALNLCLPM R 498- 505- 506 514 CMV RR GVTPAGLIGV RR GALQIBLPL R 498- 505- 506 514 HPV RR VDIRTLEDLL RR GTLGIVCPIG R 74- 84- 83 93

(53) As used herein the one-letter-code Nle refers to the non-natural amino acid norleucine.

(54) TABLE-US-00002 TABLE2 Specificpeptidesnotpartofthepresentinvention Antigen X.sup.1 X.sup.2 X.sup.3 X.sup.4 X.sup.5 HCV R GYIPLVGAPLG RRR VARALAHGVRV R HCV R GYLPAVGAPIG RRR VIRVIAHGLRL R HCV RR GYIPLVGAPLG RR VARALAHGVRV HCV RR GYIPLVGAPLG RRR VARALAHGVRV HCV RR SRNLGKVIDTLTC RR LMGYIPLVGA HCV RR GGGQIVGGVYLLP RR GPRLGVRATR HCV W GYIPLVGAPLG RR VARALAHGVRV HCV RR IRNLGRVIETLTLNleGYIPLIGA RR IRNLGRVIETLTLNleGYIPLIGA R Flu BR TAYERNleCNIL BRGR FQTVVQBA cmv R NLVPMVATV BRR NLVPMVATV B

(55) As used herein the one-letter-code Z or Nle refers to the non-natural amino acid norleucine.

(56) Antigens

(57) The specific natural antigen used in the peptide constructs according to the present invention may be a protein or peptide sequence derived from any B cell antigen, such as from any disease antigen, such as an infectious agent. Suitable antigens to be used according to the present invention include antigens derived from a bacteria, a mycobacterium, a virus, a parasite such as protozoa, a fungus, a cancer antigen, such as an oncogene, such as a thelomerase, a prion, an atopic disease antigen, an addictive or abused substance or a toxin or an antigen of an autoimmune disease, such as rheumatoid arthritis, insulin dependent diabetes, multiple sclerosis and the like.

(58) As used herein a disease antigen refers to any antigen confirmed or suspected to be involved in a specific disease.

(59) In some embodiments, the antigen is an abused or addictive substance or a portion thereof, including, but are not limited to, nicotine, a narcotic, a cough suppressant, a tranquilizer, and a sedative. In some embodiments, the antigen is a toxin, such as a toxin from a chemical weapon or natural sources, or a pollutant.

(60) Examples of bacteria for which antigens may be provided include, but are not limited to, M. tuberculosis, Mycobacterium, mycoplasma, neisseria and legionella. Examples of parasites include, but are not limited to, rickettsia and chlamydia.

(61) Examples of an infectious disease antigen is TbH9 (also known as Mtb 39A), a tuberculosis antigen. Other tuberculosis antigens include, but are not limited to DPV (also known as Mtb8.4), 381, Mtb41, Mtb40, Mtb32A, M9.9A, Mtb9.8, Mtbl, Mtb72f, Mtb59f, Mtb88f, Mtb71f, Mtb46f and Mtb31f (f indicates that it is a fusion or two or more proteins).

(62) Examples of cancer antigens may be a tumor associated antigen such as HER2, HER3 or HER4 receptor or one or more tumor-associated antigens or cell-surface receptors disclosed in US Publication No. 20080171040 or US Publication No. 20080305044 and are incorporated in their entirety by reference.

(63) Other suitable cancer antigens that may be used by the present invention include CD proteins such as CD2, CD3, CD4, CD5, CD6, CD8, CD11, CD14, CD18, CD19, CD22, CD21, CD22, CD25, CD26, CD27, CD28, CD30, CD33, CD36, CD37, CD38, CD40, CD44, CD52, CD55, CD56, CD70, CD79, CD80, CD81, CD103, CD105, CD134, CD137, CD138, and CD152; members of the ErbB receptor family such as the EGF receptor, HER2, HER3 or HER4 receptor; cell adhesion molecules such as LFA-I, Mac1, pi 50.95, VLA-4, ICAM-1, VCAM, EpCAM, alpha4/beta7 integrin, and alpha v/beta3 integrin including either alpha or beta subunits thereof (e.g. anti-CD11a, anti-CD18 or anti-CD11b antibodies); growth factors such as VEGF; tissue factor (TF); TGF-.; alpha interferon (alpha-IFN); an interleukin, such as IL-8; IgE; blood group antigens Apo2, death receptor; flk2/flt3 receptor; obesity (OB) receptor; mpl receptor; CTLA-4; protein C etc. In some embodiment the antigen is selected from IGF-IR, CanAg, EphA2, MUC1, MUC16, VEGF, TF, CD19, CD20, CD22, CD27, CD33, CD37, CD38, CD40, CD44, CD56, CD138, CA6, Her2/neu, EpCAM, CRIPTO (a protein produced at elevated levels in a majority of human breast cancer cells), darpins, alpha.sub.v/beta.sub.3 integrin, alpha.sub.v/beta.sub.5 integrin, alpha y/beta integrin, TGF-, CD11a, CD18, Apo2 and C242. In some embodiment the antigen is selected from a CD proteins such as CD3, CD4, CD8, CD19, CD20, CD27, CD34, CD37, CD38, CD46, CD56, CD70 and CD138; members of the ErbB receptor family such as the EGF receptor, HER2, HER3 or HER4 receptor; cell adhesion molecules such as LFA-I, Mac1, p150.95, VLA-4, ICAM-1, VCAM, EpCAM, alpha4/beta7 integrin, and alpha v/beta3 integrin including either alpha or beta subunits thereof (e.g. anti-CD11a, anti-CD18 or anti-CD11b antibodies); growth factors such as VEGF; tissue factor (TF); TGF-.; alpha interferon (alpha-IFN); an interleukin, such as IL-8; IgE; blood group antigens Apo2, death receptor; flk2/flt3 receptor; obesity (OB) receptor; mpl receptor; CTLA-4; protein C, etc. The most preferred targets herein are IGF-IR, CanAg, EGF-R, EGF-RvIII, EphA2, MUC1, MUC16, VEGF, TF, CD19, CD20, CD22, CD27, CD33, CD37, CD38, CD40, CD44, CD56, CD70, CD138, CA6, Her2/neu, CRIPTO (a protein produced at elevated levels in a majority of human breast cancer cells), alpha.sub.v/beta.sub.3 integrin, alpha.sub.v/beta.sub.5 integrin, TGF-, CD11a, CD18, Apo2, EpCAM and C242. In some embodiment the antigen is selected from a cellular oncogene, such as ras or myc.

(64) Examples of viral antigens for use with the present invention include, but are not limited to, e.g., HIV, HCV, CMV, HPV, Influenza, adenoviruses, retroviruses, picornaviruses, etc. Non-limiting example of retroviral antigens such as retroviral antigens from the human immunodeficiency virus (HIV) antigens such as gene products of the gag, pol, and env genes, the Nef protein, reverse transcriptase, and other HIV components; hepatitis viral antigens such as the S, M, and L proteins of hepatitis B virus, the pre-S antigen of hepatitis B virus, and other hepatitis, e.g., hepatitis A, B, and C, viral components such as hepatitis C viral RNA; influenza viral antigens such as hemagglutinin and neuraminidase and other influenza viral components; measles viral antigens such as the measles virus fusion protein and other measles virus components; rubella viral antigens such as proteins E1 and E2 and other rubella virus components; rotaviral antigens such as VP7sc and other rotaviral components; cytomegaloviral antigens such as envelope glycoprotein B and other cytomegaloviral antigen components; respiratory syncytial viral antigens such as the RSV fusion protein, the M2 protein and other respiratory syncytial viral antigen components; herpes simplex viral antigens such as immediate early proteins, glycoprotein D, and other herpes simplex viral antigen components; varicella zoster viral antigens such as gpl, gpll, and other varicella zoster viral antigen components; Japanese encephalitis viral antigens such as proteins E, M-E, M-E-NSI, NSI, NS1-NS2A, 80% E, and other Japanese encephalitis viral antigen components; rabies viral antigens such as rabies glycoprotein, rabies nucleoprotein and other rabies viral antigen components. See Fundamental Virology, Second Edition, eds. Fields, B. N. and Knipe, D. M. (Raven Press, New York, 1991) for additional examples of viral antigens.

(65) The epitopes to be incorporated into the scaffold design according to the present invention may be derived from an adenovirus, retrovirus, picornavirus, herpesvirus, rotavirus, hantavirus, coronavirus, togavirus, flavirvirus, rhabdovirus, paramyxovirus, orthomyxovirus, bunyavirus, arenavirus, reovirus, papilomavirus, parvovirus, poxvirus, hepadnavirus, degngue virus, or spongiform virus. In certain specific, non-limiting examples, the viral antigen are peptides obtained from at least one of HIV, CMV, hepatitis A, B, and C, influenza, measles, polio, smallpox, rubella; respiratory syncytial, herpes simplex, varicella zoster, Epstein-Barr, Japanese encephalitis, rabies, Influenza, and/or cold viruses.

(66) HCV:

(67) Peptides according to the present invention may comprise a known antigen. For antigens derived from HCV these antigens may be derived from the Core, E1, E2, P7, NS2, NS3, NS4 (NS4A and NS4B) and NS5 (NS5A and NS5B) protein of the Hepatitis C Virus (HCV). The epitopes are those which elicit a HLA class I and/or class II restricted T lymphocyte response in an immunized host. More specific, the HLA class I restricted peptides of the present invention may bind to at least one HLA molecule of the following HLA class I groups: HLA-A*01, HLA-A*02, HLA-A*03, HLA-A*11, HLA-A*24, HLA-B*07, HLA-B*08, HLA-B*35, HLA-B*40, HLA-B*44, HLA-Cw3, HLA-Cw4, HLA-Cw6 or HLA-Cw7. The HLA class II restricted peptides of the present invention may bind to at least one HLA molecule of the following HLA class II groups: HLA-DRB1, -DRB2, -DRB3, -DRB4, -DRB5, -DRB6, -DRB7, -DRB8 or -DRB9.

(68) MHC binding HCV peptides that may be used according to the present invention as epitopes are disclosed in e.g. WO02/34770 (Imperial College Innovations Ltd), WO01/21189 and WO02/20035 (Epimmune), WO04/024182 (Intercell), WO95/25122 (The Scripps Research Institute), WO95/27733 (Government of the USA, Department of Health and Human Services), EP 0935662 (Chiron), WO02/26785 (Immusystems GmbH), WO95/12677 (Innogenetics N.V), WO97/34621 (Cytel Corp), and EP 1652858 (Innogenetics N.V.).

(69) In other embodiments, the scaffold design according to the present invention comprises a PADRE peptide, such as the universal T cell epitope called PADRE as disclosed in WO95/07707 (Epimmune) the content of which are enclosed herein by reference. A PanDR binding peptide or PADRE peptide is a member of a family of molecules that binds more that one HLA class II DR molecule. PADRE binds to most HLA-DR molecules and stimulates in vitro and in vivo human helper T lymphocyte (HTL) responses. Alternatively T-help epitopes can be used from universally used vaccines such as tetanos toxoid.

(70) In a further embodiment, the peptides in the composition or polyepitopic peptide are characterized in that they are derived from a HCV protein, or more specifically from at least one of the following HCV regions selected from the group consisting of Core, E1, E2/NS1, NS2, NS3, NS4A, NS4B, NS5A and NS5B. Even more preferred is that peptides are characterized in that they are present in the HCV consensus sequence of genotype 1a, 1b and/or 3 a.

(71) Other HLA class I and II binding peptides that may be used according to the invention may be identified by the method as described in WO03/105058-Algonomics, by the method as described by Epimmune in WO01/21189 and/or by three public database prediction servers, respectively Syfpeithi, BIMAS and nHLAPred. It is also an aspect of this present invention that each peptide may be used within the scaffold design of the invention in combination with the same peptide as multiple repeats, or with any other peptide(s) or epitope(s).

(72) TABLE-US-00003 TABLE3 SpecificHCVpeptidesintheircompletelengthaccording totheinvention: Series Ep.nr Ver. Scaf. Z1 Z2 Z3 Z4 Z5 Z6 Z7 Z8 Z9 BI330 72 RR GGQLIGGIYLIPG RR VITFSIYLIVS BI330 72 b RRR GGQLIGGIYLIPG RR VITFSIYLIVS BI330 72 c RR GGQLIGGIYLIPG RRR VITFSIYLIVS BI330 72 d RR GGQLIGGIYLIPG RR VITFSIYLIVS R BI330 72 e RR GGQLIGGIYLIPG RR VITFSIYLIVS RR BI330 72 2 RR VITYSIFLIVS RR GGNVIGGIYZIPR BI330 72 2 b RRR VITYSIFLIVS RR GGNVIGGIYZIPR BI330 72 2 c RR VITYSIFLIVS RRR GGNVIGGIYZIPR BI330 72 2 d RRR VITYSIFLIVS RRR GGNVIGGIYZIPR Z= Nle BI330 83 RRG TANWARVIS R ANWAKVIL R NWAKVI BI330 83 b RG TANWARVIS RR ANWAKVIL R NWAKVI BI330 83 c RG TANWARVIS R ANWAKVIL R NWAKVI BI330 83 d RG TANWARVIS RG ANWAKVIL R NWAKVI BI330 83 2 RRG TANWARVIS R ANWARVIL R NWAKVI BI330 83 2 b RG TANWARVIS RR ANWARVIL R NWAKVI BI330 83 2 c RG TANWARVIS R ANWARVIL R NWAKVI BI330 83 2 d RG TANWARVIS RG ANWARVIL R NWAKVI BI310 511 R GYLPAVGAPI RRR VIRVIAHGLRL R BI310 511 b RR GYLPAVGAPI RR VIRVIAHGLRL R BI310 511 c RR GYLPAVGAPI RRR VIRVIAHGLRL BI310 511 d RR GYLPAVGAPI RR VIRVIAHGLRL BI310 511 e R GYLPAVGAPI RR VIRVIAHGLRL R BI310 511 f R GYLPAVGAPI R VIRVIAHGLRL R BI310 511 g R GYLPAVGAPI RR VIRVIAHGLRL = no amino acid; B = Cit; Z = Nle; X = Har
CMV:

(73) The epitopes to be incorporated into the scaffold design according to the present invention may be derived from cytomegalovirus (CMV) including CMV glycoproteins gB and gH.

(74) TABLE-US-00004 TABLE4 SpecificCMVpeptidesintheircompletelengthaccording totheinvention: Series Nr Ver. Scaf. Z1 Z2 Z3 Z4 Z5 Z6 Z7 Z8 BI050 4 RG NIVPZVVTA RR IGDLIVAQV BI050 4 b RR NIVPZVVTA RR IGDLIVAQV BI050 4 c RRR NIVPZVVTA RR IGDLIVAQV BI050 4 d RR NIVPZVVTA RRR IGDLIVAQV BI050 4 2 RG NIVPZVVTA RR IGDLIVQAV BI050 4 2 b RR NIVPZVVTA RR IGDLIVQAV BI050 4 2 c RRR NIVPZVVTA RR IGDLIVQAV BI050 4 2 d RR NIVPZVVTA RRR IGDLIVQAV BI050 5 RG VTPADLIGA RR QYNPVAVZF BI050 5 b RR VTPADLIGA RR QYNPVAVZF BI050 5 c RRR VTPADLIGA RR QYNPVAVZF BI050 5 d RR VTPADLIGA RRR QYNPVAVZF BI050 6 RRG PRPEGYTLFF R GYTLFFTS R BI050 6 b RG PRPEGYTLFF RR GYTLFFTS R BI050 6 c RRG PRPEGYTLFF RR GYTLFFTS R BI050 6 d RRG PRPEGYTLFF RRR GYTLFFTS R BI050 6 e RRRG PRPEGYTLFF RR GYTLFFTS R BI050 7 RG LPYPRGYTLFV RR GYTLFVSD R BI050 7 b RRG LPYPRGYTLFV RR GYTLFVSD R BI050 7 c RRG LPYPRGYTLFV RRR GYTLFVSD R BI050 7 d RRRG LPYPRGYTLFV RR GYTLFVSD R BI050 7 e RRG LPYPRGYTLFV RR GYTLFVSD R BI050 8 RRG ETILTPRDV R NTLZTPRDV R BI050 8 b RG ETILTPRDV RR NTLZTPRDV R BI050 8 c RG ETILTPRDV R NTLZTPRDV R BI050 8 d RG ETILTPRDV RG NTLZTPRDV R BI050 9 RR SSTSPVYDL RR SSTSPVYNL R BI050 9 b RR SSTSPVYDL RRR SSTSPVYNL R BI050 9 c RRR SSTSPVYDL RR SSTSPVYNL R BI050 9 d RRR SSTSPVYDL RRR SSTSPVYNL R = no amino acid; B = Cit; Z = Nle; X = Har
Influenza:

(75) The epitopes to be incorporated into the scaffold design according to the present invention may be derived from fragments or portions of Influenza hemagglutinin (HA) or Influenza neuraminidase (NA), nucleoprotein (NP), M1, M2, NS1, NEP, PA, PB1, PB1-F2, PB2 for each of the subgroups, such as H1N1, H2N2 og H3N2.

(76) Suitable epitopes may be derived from an HA protein of one, or more than one subtype, including H1, H2, H3, H4, H5, H6, H7, H8, H9, H10, H11, H12, H13, H14, H15 or H16 or fragment or portion thereof. Examples of subtypes comprising such HA proteins include A/New Calcdonia/20/99 (H1N1) A/Indonesia/5/2006 (H5N1), A/chicken/New York/1995, A/herring gull/DE/677/88 (H2N8), A/Texas/32/2003, A/mallard/MN/33/00, A/duck/Shanghai/1/2000, A/northern pintail/TX/828189/02, A/Turkey/Ontario/6118/68 (H8N4), A/shoveler/Iran/G54/03, A/chicken/Germany/N/1949 (H10N7), A/duck/England/56 (H11N6), A/duck/Alberta/60/76 (H12N5), A/Gull/Maryland/704/77 (H13N6), A/Mallard/Gurjev/263/82, A/duck/Australia/341/83 (H15N8), A/black-headed gull/Sweden/5/99 (H16N3), B/Lee/40, C/Johannesburg/66, A/PuertoRico/8/34 (H1N1), A/Brisbane/59/2007 (H1N1), A/Solomon Islands 3/2006 (H1N1), A/Brisbane 10/2007 (H3N2), A/Wisconsin/67/2005 (H3N2), B/Malaysia/2506/2004, B/Florida/4/2006, A/Singapore/1/57 (H2N2), A/Anhui/1/2005 (H5N1), A/Vietnam/1194/2004 (H5N1), A/Teal/HongKong/W312/97 (H6N1), A/Equine/Prague/56 (H7N7), A/HongKong/1073/99 (H9N2)).

(77) In some embodiments of the invention, the HA protein may be an H1, H2, H3, H5, H6, H7 or H9 subtype. In other embodiments, the H1 protein may be from the A/New Calcdonia/20/99 (H1N1), A/PuertoRico/8/34 (H1N1), A/Brisbane/59/2007 (H1N1), or A/Solomon Islands 3/2006 (H1N1) strain. The H3 protein may also be from the A/Brisbane 10/2007 (H3N2) or A/Wisconsin/67/2005 (H3N2) strain. In other embodiments, the H2 protein may be from the A/Singapore/1/57 (H2N2) strain. The H5 protein may be from the A/Anhui/1/2005 (H5N1), A/Vietnam/1194/2004 (H5N1), or A/Indonesia/5/2005 strain. In other embodiments, the H6 protein may be from the A/Teal/HongKong/W312/97 (H6N1) strain. The H7 protein may be from the A/Equine/Prague/56 (H7N7) strain. In other embodiments, the H9 protein is from the A/HongKong/1073/99 (H9N2) strain. In other embodiments, the HA protein may be from an influenza virus may be a type B virus, including B/Malaysia/2506/2004 or B/Florida/4/2006. The influenza virus HA protein may be H5 Indonesia.

(78) TABLE-US-00005 TABLE5 SpecificInfluenzapeptidesaccordingtotheinventionintheir completelength(ZorNledenotesNorleucine,XorHardenotes homoarginine): Series Ep.nr version scaffold Z1 Z2 Z3 Z4 Z5 Z6 Z7 BI100 330 RR TAYERZCNIL RR GLEPLVIAGILA BI100 330 b RRR TAYERZCNIL RR GLEPLVIAGILA BI100 330 c RR TAYERZCNIL RRR GLEPLVIAGILA BI100 330 d RR TAYERZCNIL RR GLEPLVIAGILA R BI100 330 e RR TAYERZCNIL RR GLEPLVIAGILA RR Z= Nle BI100 270 RR TVIGASZIPLL RG TPIXQDWENRAN BI100 270 b RRR TVIGASZIPLL RG TPIXQDWENRAN BI100 270 c RR TVIGASZIPLL RRG TPIXQDWENRAN BI100 270 d RRR TVIGASZIPLL RRG TPIXQDWENRAN BI100 270 e RRR TVIGASZIPLL RRG TPIXQDWENRAN R Z= Nle X= Har BI100 130 RR AAFEEZXITS RR VAFEDLXZZSFI BI100 130 b RRR AAFEEZXITS RR VAFEDLXZZSFI BI100 130 c RRR AAFEEZXITS RRG VAFEDLXZZSFI BI100 130 d RRR AAFEEZXITS RRR VAFEDLXZZSFI BI100 130 e RRR AAFEEZXITS RRR VAFEDLXZZSFI GR BI100 190 e RR TAYERZCNIL RRG RFQTVVQBA BI100 190 f RR TAYERZCNIL RRG RFQTVVQBA R BI100 190 g R TAYERZCNIL RG RFQTVVQBA R BI100 190 h RR TAYERZCNIL RG RFQTVVQBA BI100 260 b BR GLEPLVIAGILA RR GSLVGLLHIVL BI100 260 c RR GLEPLVIAGILA RR GSLVGLLHIVL BI100 260 d RR GLEPLVIAGILA RR GSLVGLLHIVL R BI100 260 e RR GLEPLVIAGILA RRR GSLVGLLHIVL BI100 260 f RR GLEPLVIAGILA RRR GSLVGLLHIVL R BI100 120 3 a R TAFLVRNVA R SIARSVTIZXASVVH BI100 120 3 b R TAFLVRNVA RR SIARSVTIZXASVVH BI100 120 3 c RR TAFLVRNVA R SIARSVTIZXASVVH BI100 120 3 d RR TAFLVRNVA RR SIARSVTIZXASVVH BI100 120 3 e RR TAFLVRNVA RR SIARSVTIZXASVVH R BI100 120 3 f RR TAFLVRNVA RR SIARSVTIZXASVVH RR BI100 220 RG Dpr TPI(Har)QDWGN RG TPTRQEWDCRIS (Aoa) RAN BI100 220 2 RG Dpr TPI(Har)QDWGN RG TPTRQEWDARIS (Aoa) RAN BI100 220 3 RG TPI(Har)QDWGN RG TPTRQEWDCRIS RAN BI100 220 4 RG TPI(Har)QDWGN RG TPTRQEWDARIS RAN BI100 220 5 RG C TPI(Har)QDWGN RG TPTRQEWDCRIS RAN BI100 220 6 RG C TPI(Har)QDWGN RG TPTRQEWDARIS RAN BI100 220 7 RG K TPI(Har)QDWGN RG TPTRQEWDCRIS RAN BI100 220 8 RG K TPI(Har)QDWGN RG TPTRQEWDARIS RAN BI100 220 9 RG Lys TPI(Har)QDWGN RG TPTRQEWDCRIS (Me) RAN BI100 220 10 RG Lys TPI(Har)QDWGN RG TPTRQEWDARIS (Me) RAN BI100 220 11 RG D TPI(Har)QDWGN RG TPTRQEWDCRIS RAN BI100 220 12 RG D TPI(Har)QDWGN RG TPTRQEWDARIS RAN BI100 220 13 RG E TPI(Har)QDWGN RG TPTRQEWDCRIS RAN BI100 220 14 RG E TPI(Har)QDWGN RG TPTRQEWDARIS RAN BI100 240 RG Dpr TPT(Har)NGWDV RG TPI(Har)QEW(Har)SL (Ser) KLS (Nle)NQEW BI100 240 3 RG TPT(Har)NGWDV RG TPI(Har)QEW(Har)SL KLS (Nle)NQEW BI100 240 4 RG K TPT(Har)NGWDV RG TPI(Har)QEW(Har)SL KLS (Nle)NQEW BI100 240 5 RG C TPT(Har)NGWDV RG TPI(Har)QEW(Har)SL KLS (Nle)NQEW BI100 240 6 RG Lys TPT(Har)NGWDV RG TPI(Har)QEW(Har)SL (Me) KLS (Nle)NQEW BI100 240 7 RG D TPT(Har)NGWDV RG TPI(Har)QEW(Har)SL KLS (Nle)NQEW BI100 240 8 RG E TPT(Har)NGWDV RG TPI(Har)QEW(Har)SL KLS (Nle)NQEW = no amino acid; B = Cit; Z = Nle; X = Har

(79) TABLE-US-00006 TABLE6 SpecificdimericInfluenzapeptidesaccordingtotheinvention intheircompletelength(ZorNledenotesNorleucine,Xor Hardenoteshomoarginine,residueslinkingAandBmonomer peptidestodimersareunderlined): Dimeric Dimericpeptides,composedofpeptides Constituent Peptide AandB.Linkedresiduesunderlined monomers BI-155 A RG(Dpr(Aoa))-TPI(Har)QDWGNRAN-RG- BI-100-220 TPTRQEWDCRIS-NH2 B RG(Dpr(Ser))-TPT(Har)NGWDVKLS-RG- BI-100-240 TPI(Har)QEW(Har)SL(Nle)NQEW-NH2 BI-155-2 A RG(Dpr(Aoa))-TPI(Har)QDWGNRAN-RG- BI-100-220-2 TPTRQEWDARIS-NH2 B RG(Dpr(Ser))-TPT(Har)NGWDVKLS-RG- BI-100-240 TPI(Har)QEW(Har)SL(Nle)NQEW-NH2 BI-155-3 A RGC-TPI(Har)QDWGNRAN-RG-TPTRQEWDCRIS- BI-100-220-5 NH2 B RGK-TPT(Har)NGWDVKLS-RG- BI-100-240-4 TPI(Har)QEW(Har)SL(Nle)NQEW-NH2 BI-155-4 A RGC-TPI(Har)QDWGNRAN-RG-TPTRQEWDARIS- BI-100-220-6 NH2 B RGK-TPT(Har)NGWDVKLS-RG- BI-100-240-4 TPI(Har)QEW(Har)SL(Nle)NQEW-NH2 BI-155-5 A RGK-TPI(Har)QDWGNRAN-RG-TPTRQEWDCRIS- BI-100-220-7 NH2 B RGC-TPT(Har)NGWDVKLS-RG- BI-100-240-5 TPI(Har)QEW(Har)SL(Nle)NQEW-NH2 BI-155-6 A RGK-TPI(Har)QDWGNRAN-RG-TPTRQEWDARIS- BI-100-220-8 NH2 B RGC-TPT(Har)NGWDVKLS-RG- BI-100-240-5 TPI(Har)QEW(Har)SL(Nle)NQEW-NH2 BI-155-7 A RG(Lys(Me))-TPI(Har)QDWGNRAN-RG- BI-100-220-9 TPTRQEWDCRIS-NH2 B RGD-TPT(Har)NGWDVKLS-RG- BI-100-240-7 TPI(Har)QEW(Har)SL(Nle)NQEW-NH2 BI-155-8 A RG(Lys(Me))-TPI(Har)QDWGNRAN-RG- BI-100-220-9 TPTRQEWDCRIS-NH2 B RGE-TPT(Har)NGWDVKLS-RG- BI-100-240-8 TPI(Har)QEW(Har)SL(Nle)NQEW-NH2 BI-155-9 A RG(Lys(Me))-TPI(Har)QDWGNRAN-RG- BI-100-220-10 TPTRQEWDARIS-NH2 B RGD-TPT(Har)NGWDVKLS-RG- BI-100-240-7 TPI(Har)QEW(Har)SL(Nle)NQEW-NH2 BI-155-10 A RG(Lys(Me))-TPI(Har)QDWGNRAN-RG- BI-100-220-10 TPTRQEWDARIS-NH2 B RGE-TPT(Har)NGWDVKLS-RG- BI-100-240-8 TPI(Har)QEW(Har)SL(Nle)NQEW-NH2 BI-155-11 A RGD-TPI(Har)QDWGNRAN-RG-TPTRQEWDCRIS- BI-100-220-11 NH2 B RG(Lys(Me))-TPT(Har)NGWDVKLS-RG- BI-100-240-6 TPI(Har)QEW(Har)SL(Nle)NQEW-NH2 BI-155-12 A RGD-TPI(Har)QDWGNRAN-RG-TPTRQEWDARIS- BI-100-220-12 NH2 B RG(Lys(Me))-TPT(Har)NGWDVKLS-RG- BI-100-240-6 TPI(Har)QEW(Har)SL(Nle)NQEW-NH2 BI-155-13 A RGE-TPI(Har)QDWGNRAN-RG-TPTRQEWDCRIS- BI-100-220-13 NH2 B RG(Lys(Me))-TPT(Har)NGWDVKLS-RG- BI-100-240-6 TPI(Har)QEW(Har)SL(Nle)NQEW-NH2 BI-155-14 A RGE-TPI(Har)QDWGNRAN-RG-TPTRQEWDARIS- BI-100-220-14 NH2 B RG(Lys(Me))-TPT(Har)NGWDVKLS-RG- BI-100-240-6 TPI(Har)QEW(Har)SL(Nle)NQEW-NH2 BI-155-15 A RGC-TPI(Har)QDWGNRAN-RG-TPTRQEWDCRIS- BI-100-220-5 NH2 B RGC-TPT(Har)NGWDVKLS-RG- BI-100-240-5 TPI(Har)QEW(Har)SL(Nle)NQEW-NH2 BI-155-16 A RGC-TPI(Har)QDWGNRAN-RG-TPTRQEWDARIS- BI-100-220-6 NH2 B RGC-TPT(Har)NGWDVKLS-RG- BI-100-240-5 TPI(Har)QEW(Har)SL(Nle)NQEW-NH2 A-monomerpeptidevariants: RG(Dpr(Aoa))-TPI(Har)QDWGNRAN-RG- BI-100-220 TPTRQEWDCRIS-NH2 RG(Dpr(Aoa))-TPI(Har)QDWGNRAN-RG- BI-100-220-2 TPTRQEWDARIS-NH2 RG-TPI(Har)QDWGNRAN-RG-TPTRQEWDCRIS-NH2 BI-100-220-3 RG-TPI(Har)QDWGNRAN-RG-TPTRQEWDARIS-NH2 BI-100-220-4 RGC-TPI(Har)QDWGNRAN-RG-TPTRQEWDCRIS- BI-100-220-5 NH2 RGC-TPI(Har)QDWGNRAN-RG-TPTRQEWDARIS- BI-100-220-6 NH2 RGK-TPI(Har)QDWGNRAN-RG-TPTRQEWDCRIS- BI-100-220-7 NH2 RGK-TPI(Har)QDWGNRAN-RG-TPTRQEWDARIS- BI-100-220-8 NH2 RG(Lys(Me))-TPI(Har)QDWGNRAN-RG- BI-100-220-9 TPTRQEWDCRIS-NH2 RG(Lys(Me))-TPI(Har)QDWGNRAN-RG- BI-100-220-10 TPTRQEWDARIS-NH2 RGD-TPI(Har)QDWGNRAN-RG-TPTRQEWDCRIS- BI-100-220-11 NH2 RGD-TPI(Har)QDWGNRAN-RG-TPTRQEWDARIS- BI-100-220-12 NH2 RGE-TPI(Har)QDWGNRAN-RG-TPTRQEWDCRIS- BI-100-220-13 NH2 RGE-TPI(Har)QDWGNRAN-RG-TPTRQEWDARIS- BI-100-220-14 NH2 B-monomerpeptidevariants: RG(Dpr(Ser))-TPT(Har)NGWDVKLS-RG- BI-100-240 TPI(Har)QEW(Har)SL(Nle)NQEW-NH2 RG-TPT(Har)NGWDVKLS-RG- BI-100-240-3 TPI(Har)QEW(Har)SL(Nle)NQEW-NH2 RGK-TPT(Har)NGWDVKLS-RG- BI-100-240-4 TPI(Har)QEW(Har)SL(Nle)NQEW-NH2 RGC-TPT(Har)NGWDVKLS-RG- BI-100-240-5 TPI(Har)QEW(Har)SL(Nle)NQEW-NH2 RG(Lys(Me))-TPT(Har)NGWDVKLS-RG- BI-100-240-6 TPI(Har)QEW(Har)SL(Nle)NQEW-NH2 RGD-TPT(Har)NGWDVKLS-RG- BI-100-240-7 TPI(Har)QEW(Har)SL(Nle)NQEW-NH2 RGE-TPT(Har)NGWDVKLS-RG- BI-100-240-8 TPI(Har)QEW(Har)SL(Nle)NQEW-NH2
Human Immunodeficiency Virus (HIV):

(80) For HIV, the epitopes to be incorporated into the scaffold design according to the present invention may be derived from the group consisting of gp120, gp160, gp41, p24gag or p55gag derived from HIV, including members of the various genetic subtypes.

(81) Human Papillomavirus (HPV):

(82) For HPV, the epitopes to be incorporated into the scaffold design according to the present invention may be derived from the group consisting E1, E2, E3, E4, E6 and E7, L1 and L2 proteins. The epitopes may be derived from any type including types 8, 11, 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, and 59.

(83) TABLE-US-00007 TABLE7 SpecificHPVpeptidesintheircompletelengthaccording totheinvention: Series Nr Version Scaffold Z1 Z2 Z3 Z4 Z5 Z6 Z7 Z8 Native: 35-45 48-58 BI500 1 RR LECVYCKQQLL RR EVYDFAFRDL C BI500 1 b RR LECVYCKQQLL RRR EVYDFAFRDL G C BI500 1 c RRR LECVYCKQQLL RRG EVYDFAFRDL C BI500 1 d RRR LECVYCKQQLL RRR EVYDFAFRDL G C BI500 1 e RRRG LECVYCKQQLL RRR EVYDFAFRDL G C Native: 49-58 52-61 BI500 2 RR GVYDFAFRDLC RR GFAFRDLCIV R Y BI500 2 b RR GVYDFAFRDLC RRR GFAFRDLCIV G Y BI500 2 c RRR GVYDFAFRDLC RRG GFAFRDLCIV R Y BI500 2 d RRR GVYDFAFRDLC RRR GFAFRDLCIV G Y BI500 2 e RRRG GVYDFAFRDLC RRR GFAFRDLCIV R G Y Native: 49-58 52-61 BI500 3 RR GVFDYAFRDIN RR GFAYRDINLA R Y BI500 3 b RR GVYDFAFRDLC RRR GFAFRDLCIV G Y BI500 3 c RRR GVYDFAFRDLC RRG GFAFRDLCIV R Y BI500 3 d RRR GVYDFAFRDLC RRR GFAFRDLCIV G Y BI500 3 e RRRG GVYDFAFRDLC RRR GFAFRDLCIV R G Y Native: 74-83 84-93 BI500 4 RR VDIRTLEDLL RR GTLGIVCPIG R BI500 4 b RR VDIRTLEDLL RRR GTLGIVCPIG G BI500 4 c RRR VDIRTLEDLL RRG GTLGIVCPIG R BI500 4 d RRR VDIRTLEDLL RRR GTLGIVCPIG G BI500 4 e RRRG VDIRTLEDLL RRR GTLGIVCPIG R G

(84) The present invention further relates to compositions comprising two or three peptides of the invention.

(85) TABLE-US-00008 TABLE8 Thetablerepresent10differentsuitablecombinationsofthree monomericpeptideseachpeptidecomprisingaspecificnatural antigenofaproteinorpeptidesequencederivedfromHCV. 1 BI3 BI3 BI3 RRGGQLIGGI RRGTANWARV RRGYLPAVG (SEQID (SEQID (SEQID 30- 30- 10- YLIPGRRVITF ISRANWAKVIL APIRRVIRVI NO:357) NO:366) NO:377) 72 83 511 SIYLIVS RNWAKVI AHGLRL d 2 BI3 BI3 BI3 RRRGGQLIGG RGTANWARVI RGYLPAVGA (SEQID (SEQID (SEQID 30- 30- 10- IYLIPGRRVITF SRRANWAKVI PIRVIRVIAH NO:358) NO:367) NO:379) 72b 83b 511 SIYLIVS LRNWAKVI GLRLR f 3 BI3 BI3 BI3 RRGGQLIGGI RGTANWARVI RGYLPAVGA (SEQID (SEQID (SEQID 30- 30- 10- YLIPGRRRVIT SRANWAKVIL PIRRVIRVIA NO:359) NO:368) NO:380) 72c 83c 511 FSIYLIVS RNWAKVI HGLRL g 4 BI3 BI3 BI3 RRGGQLIGGI RGTANWARVI RGYLPAVGA (SEQID (SEQID (SEQID 30- 30- 10- YLIPGRRVITF SRGANWAKVI PIRRRVIRVI NO:360) NO:369) NO:374) 72d 83d 511 SIYLIVSR LRNWAKVI AHGLRLR 5 BI3 BI3 BI3 RRGGQLIGGI RRGTANWARV RRGYLPAVG (SEQID (SEQID (SEQID 30- 30- 10- YLIPGRRVITF ISRANWARVIL APIRRVIRVI NO:361) NO:370) NO:375) 72e 83- 511 SIYLIVSRR RNWAKVI AHGLRLR 2 b 6 BI3 BI3 BI3 RRVITYSIFLIV RGTANWARVI RRGYLPAVG (SEQID (SEQID (SEQID 30- 30- 10- SRRGGNVIGG SRRANWARVI APIRRRVIRV NO:362) NO:371) NO:376) 72- 83- 511 IYZIPR LRNWAKVI IAHGLRL 2 2b c 7 BI3 BI3 BI3 RRVITYSIFLIV RRGTANWARV RRGYLPAVG (SEQID (SEQID (SEQID 30- 30- 10- SRRGGNVIGG ISRANWAKVIL APIRRVIRVI NO:362) NO:366) NO:377) 72- 83 511 IYZIPR RNWAKVI AHGLRL 2 d 8 BI3 BI3 BI3 RRRVITYSIFLI RGTANWARVI RGYLPAVGA (SEQID (SEQID (SEQID 30- 30- 10- VSRRGGNVIG SRRANWAKVI PIRRVIRVIA NO:363) NO:367) NO:378) 72- 83b 511 GIYZIPR LRNWAKVI HGLRLR 2b e 9 BI3 BI3 BI3 RRVITYSIFLIV RGTANWARVI RGYLPAVGA (SEQID (SEQID (SEQID 30- 30- 10- SRRRGGNVIG SRANWARVIL PIRVIRVIAH NO:364) NO:372) NO:379) 72- 83- 511 GIYZIPR RNWAKVI GLRLR 2c 2c f 10 BI3 BI3 BI3 RRRVITYSIFLI RGTANWARVI RGYLPAVGA (SEQID (SEQID (SEQID 30- 30- 10- VSRRRGGNVI SRGANWARVI PIRRVIRVIA NO:365) NO:373) NO:380) 72- 83- 511 GGIYZIPR LRNWAKVI HGLRL 2d 2d g

(86) TABLE-US-00009 TABLE9 Thetablerepresent10differentsuitablecombinationsof threemonomericpeptidesandonedimericpeptideeach peptidecomprisingspecificnaturalantigenofaproteinor peptidesequencederivedfrominfluenza. 1 BI- BI100-330; BI100-270 BI100-130 155- RRTAYERZCNILRRGLEP RRTVIGASZIPLLRGTPIXQD RRAAFEEZXITSRRVAFEDL 5 LVIAGILA WENRAN XZZSFI (SEQIDNO:407) (SEQIDNO:412) (SEQIDNO:417) 2 BI- BI100-330b BI100-270b BI100-130b 155- RRRTAYERZCNILRRGLE RRRTVIGASZIPLLRGTPIXQ RRRAAFEEZXITSRRVAFED 4 PLVIAGILA DWENRAN LXZZSFI (SEQIDNO:408) (SEQIDNO:413) (SEQIDNO:418) 3 BI- BI100-330c BI100-270c BI100-130c 155- RRTAYERZCNILRRRGLE RRTVIGASZIPLLRRGTPIXQ RRRAAFEEZXITSRRGVAFE 3 PLVIAGILA DWENRAN DLXZZSFI (SEQIDNO:409) (SEQIDNO:414) (SEQIDNO:419) 4 BI- BI100-330d BI100-270d BI100-130d 155- RRTAYERZCNILRRGLEP RRRTVIGASZIPLLRRGTPIX RRRAAFEEZXITSRRRVAFE 2 LVIAGILAR QDWENRAN DLXZZSFI (SEQIDNO:410) (SEQIDNO:415) (SEQIDNO:420) 5 BI- BI100-330e BI100-270e BI100-130e 155 RRTAYERZCNILRRGLEP RRRTVIGASZIPLLRRGTPIX RRRAAFEEZXITSRRRVAFE LVIAGILARR QDWENRANR DLXZZSFIGR (SEQIDNO:411) (SEQIDNO:416) (SEQIDNO:421) 6 BI- BI100-330e BI100-270e BI100-130e 155- RRTAYERZCNILRRGLEP RRRTVIGASZIPLLRRGTPIX RRRAAFEEZXITSRRRVAFE 2 LVIAGILARR QDWENRANR DLXZZSFIGR (SEQIDNO:411) (SEQIDNO:416) (SEQIDNO:421) 7 BI- BI100-330d BI100-270c BI100-130c 155- RRTAYERZCNILRRGLEP RRTVIGASZIPLLRRGTPIXQ RRRAAFEEZXITSRRGVAFE 3 LVIAGILAR DWENRAN DLXZZSFI (SEQIDNO:410) (SEQIDNO:414) (SEQIDNO:419) 10 BI- BI100-330 BI100-270d BI100-130b 155 RRTAYERZCNILRRGLEP RRRTVIGASZIPLLRRGTPIX RRRAAFEEZXITSRRVAFED LVIAGILA QDWENRAN LXZZSFI (SEQIDNO:407) (SEQIDNO:415) (SEQIDNO:418)
Carriers, Adjuvants and VehiclesDelivery

(87) The isolated peptides according to the invention may be delivered by various means and within various compositions, herein referred to as compositions, vaccine compositions or pharmaceutical compositions. The peptides of the present invention and pharmaceutical and vaccine compositions of the invention are useful for administration to mammals, particularly humans, to treat and/or prevent virus infection. Vaccine compositions containing the peptides of the invention are administered to a patient infected with the virus in question or to an individual susceptible to, or otherwise at risk for, virus infection to elicit an immune response against the specific antigens and thus enhance the patient's own immune response capabilities.

(88) Various art-recognized delivery systems may be used to deliver the peptides, into appropriate cells. The peptides can be delivered in a pharmaceutically acceptable carrier or as colloidal suspensions, or as powders, with or without diluents. They can be naked or associated with delivery vehicles and delivered using delivery systems known in the art.

(89) A pharmaceutically acceptable carrier or pharmaceutically acceptable adjuvant is any suitable excipient, diluent, carrier and/or adjuvant which, by themselves, do not induce the production of antibodies harmful to the individual receiving the composition nor do they elicit protection. Preferably, a pharmaceutically acceptable carrier or adjuvant enhances the immune response elicited by an antigen. Suitable carriers or adjuvant typically comprise one or more of the compounds included in the following non-exhaustive list: large slowly metabolized macromolecules such as proteins, polysaccharides, polylactic acids, polyglycolic acids, polymeric amino acids, amino acid copolymers and inactive virus particles; aluminium hydroxide, aluminium phosphate (see International Patent Application Publication No. WO93/24148), alum (KAI(SO4)2.12H2O), or one of these in combination with 3-0-deacylated monophosphoryl lipid A (see International Patent Application Publication No. WO93/19780); N-acetyl-muramyl-L-threonyl-D-isoglutamine (see U.S. Pat. No. 4,606,918), N-acetyl-normuramyl-L-alanyl-D-isoglutamine, N-acetylmuramyl-L-alanyl-D-isoglutamyl-L-alanine-2-(1,2-dipalmitoyl-sn-glycero-3-hydroxyphosphoryloxy)ethylamine; RIBI (ImmunoChem Research Inc., Hamilton, Mont., USA) which contains monophosphoryl lipid A (i.e., a detoxified endotoxin), trehalose-6,6-dimycolate, and cell wall skeleton (MPL+TDM+CWS) in a 2% squalene/Tween 80 emulsion. Any of the three components MPL, TDM or CWS may also be used alone or combined 2 by 2; adjuvants such as Stimulon (Cambridge Bioscience, Worcester, Mass., USA), SAF-1 (Syntex); adjuvants such as combinations between QS21 and 3-de-O-acetylated monophosphoryl lipid A (see International Application No. WO94/00153) which may be further supplemented with an oil-in-water emulsion (see, e.g., International Application Nos. WO95/17210, WO97/01640 and WO9856414) in which the oil-in-water emulsion comprises a metabolisable oil and a saponin, or a metabolisable oil, a saponin, and a sterol, or which may be further supplemented with a cytokine (see International Application No. WO98/57659); adjuvants such as MF-59 (Chiron), or poly[di(carboxylatophenoxy) phosphazene] based adjuvants (Virus Research Institute); blockcopolymer based adjuvants such as Optivax (Vaxcel, Cytrx) or inulin-based adjuvants, such as Algammulin and Gammalnulin (Anutech); Complete or Incomplete Freund's Adjuvant (CFA or IFA, respectively) or Gerbu preparations (Gerbu Biotechnik); a saponin such as QuilA, a purified saponin such as QS21, QS7 or QS17, -escin or digitonin; immunostimulatory oligonucleotides comprising unmethylated CpG dinucleotides such as [purine-purine-CG-pyrimidine-pyrimidine] oligonucleotides. These immunostimulatory oligonucleotides include CpG class A, B, and C molecules (Coley Pharmaceuticals), ISS (Dynavax), Immunomers (Hybridon). Immunostimulatory oligonucleotides may also be combined with cationic peptides as described, e.g., by Riedl et al. (2002); Immune Stimulating Complexes comprising saponins, for example Quil A (ISCOMS); excipients and diluents, which are inherently non-toxic and non-therapeutic, such as water, saline, glycerol, ethanol, isopropyl alcohol, DMSO, wetting or emulsifying agents, pH buffering substances, preservatives, and the like; a biodegradable and/or biocompatible oil such as squalane, squalene, eicosane, tetratetracontane, glycerol, peanut oil, vegetable oil, in a concentration of, e.g., 1 to 10% or 2.5 to 5%; vitamins such as vitamin C (ascorbic acid or its salts or esters), vitamin E (tocopherol), or vitamin A; carotenoids, or natural or synthetic flavanoids; trace elements, such as selenium; any Toll-like receptor ligand as reviewed in Barton and Medzhitov (2002).

(90) For a further enhancement of the vaccine antigenic properties, could be to combine a well known adjuvant with an oral immune modulant, such as IMID or adjuvant such as a Cox-2 inhibitor or a immunomodulating compound.

(91) A further aspect of the invention is the use of the vaccine combined with adjuvant, with an (oral) immunemodulating agent and a reservoir purging agent.

(92) Other suitable adjuvants includes response-selective C5a agonists, such as EP54 and EP67 described in Hung C Y et al. An agonist of human complement fragment C5a enhances vaccine immunity against Coccidioides infection. Vaccine (2012) and Kollessery G et al. Tumor-specific peptide based vaccines containing the conformationally biased, response-selective C5a agonists EP54 and EP67 protect against aggressive large B cell lymphoma in a syngeneic murine model. Vaccine (2011) 29: 5904-10.

(93) Other suitable adjuvants include an oil-in-water emulsion containing a stabilizing detergent, a micelle-forming agent and a biodegradable oil, such as Provax described in e.g. U.S. Pat. No. 5,585,103.

(94) Any of the afore-mentioned adjuvants comprising 3-de-O-acetylated monophosphoryl lipid A, said 3-de-O-acetylated monophosphoryl lipid A may be forming a small particle (see International Application No. WO94/21292).

(95) In any of the aforementioned adjuvants MPL or 3-de-O-acetylated monophosphoryl lipid A can be replaced by a synthetic analogue referred to as RC-529 or by any other amino-alkyl glucosaminide 4-phosphate (Johnson et al. 1999, Persing et al. 2002). Alternatively it can be replaced by other lipid A analogues such as OM-197 (Byl et al. 2003).

(96) A pharmaceutically acceptable vehicle includes vehicles such as water, saline, physiological salt solutions, glycerol, ethanol, etc. Auxiliary substances such as wetting or emulsifying agents, pH buffering substances, preservatives may be included in such vehicles. Delivery systems known in the art are e.g. lipopeptides, peptide compositions encapsulated in poly-DL-lactide-co-glycolide (PLG), microspheres, peptide compositions contained in immune stimulating complexes (ISCOMS), multiple antigen peptide systems (MAPs), viral delivery vectors, particles of viral or synthetic origin, adjuvants, liposomes, lipids, microparticles or microcapsules, gold particles, nanoparticles, polymers, condensing agents, polysaccharides, polyamino acids, dendrimers, saponins, QS21, adsorption enhancing materials, fatty acids or, naked or particle absorbed cDNA.

(97) The peptides may be delivered in oils such as Endocine and Montanide (Eurocine)-Montanide ISA 51 VG or Montanide ISA 720 VG (Seppic).

(98) The adjuvant may be stimulators of the innate immune system that can be given separately from the peptide such as Leukotriene B4 (LTB4) and granulocyte macrophage colony stimulating factor (GM-CSF), such as Sargramostim/Leukine (glycosylated GM-CSF) and Molgramostim (nonglycosylated GM-CSF).

(99) Typically, a vaccine or vaccine composition is prepared as an injectable, either as a liquid solution or suspension. Injection may be subcutaneous, intramuscular, intravenous, intraperitoneal, intrathecal, intradermal, or intraepidermal. Other types of administration comprise electroporation, implantation, suppositories, oral ingestion, enteric application, inhalation, aerosolization or nasal spray or drops. Solid forms, suitable for dissolving in, or suspension in, liquid vehicles prior to injection may also be prepared. The preparation may also be emulsified or encapsulated in liposomes for enhancing adjuvant effect.

(100) A liquid formulation may include oils, polymers, vitamins, carbohydrates, amino acids, salts, buffers, albumin, surfactants, or bulking agents. Preferably carbohydrates include sugar or sugar alcohols such as mono-, di-, tri-, oligo- or polysaccharides, or water-soluble glucans. The saccharides or glucans can include fructose, dextrose, lactose, glucose, mannose, sorbose, xylose, maltose, sucrose, dextran, pullulan, dextrin, alpha and beta cyclodextrin, soluble starch, hydroxethyl starch and carboxymethylcellulose, or mixtures thereof. Sucrose is most preferred. Sugar alcohol is defined as a C4 to C8 hydrocarbon having an OH group and includes galactitol, inositol, mannitol, xylitol, sorbitol, glycerol, and arabitol. Mannitol is most preferred. These sugars or sugar alcohols mentioned above may be used individually or in combination. There is no fixed limit to the amount used as long as the sugar or sugar alcohol is soluble in the aqueous preparation. Preferably, the sugar or sugar alcohol concentration is between 1.0% (w/v) and 7.0% (w/v), more preferable between 2.0 and 6.0% (w/v). Preferably amino acids include levorotary (L) forms of carnitine, arginine, and betaine; however, other amino acids may be added. Preferred polymers include polyvinylpyrrolidone (PVP) with an average molecular weight between 2,000 and 3,000, or polyethylene glycol (PEG) with an average molecular weight between 3,000 and 5,000. It is also preferred to use a buffer in the composition to minimize pH changes in the solution before lyophilization or after reconstitution. Any physiological buffer may be used, but citrate, phosphate, succinate, and glutamate buffers or mixtures thereof are preferred. Most preferred is a citrate buffer. Preferably, the concentration is from 0.01 to 0.3 molar. Surfactants that can be added to the formulation are shown in EP patent applications No. EP 0 270 799 and EP 0 268 110.

(101) Additionally, the peptides according to the present invention may be chemically modified by covalent conjugation to a polymer to increase their circulating half-life, for example. Preferred polymers, and methods to attach them to peptides, are shown in U.S. Pat. Nos. 4,766,106; 4,179,337; 4,495,285; and 4,609,546. Preferred polymers are polyoxyethylated polyols and polyethylene glycol (PEG). PEG is soluble in water at room temperature and has the general formula:

(102) R(OCH2-CH2)nOR where R can be hydrogen, or a protective group such as an alkyl or alkanol group. Preferably, the protective group has between 1 and 8 carbons, more preferably it is methyl. The symbol n is a positive integer, preferably between 1 and 1.000, more preferably between 2 and 500. The PEG has a preferred average molecular weight between 1000 and 40.000, more preferably between 2000 and 20.000, most preferably between 3.000 and 12.000. Preferably, PEG has at least one hydroxy group, more preferably it is a terminal hydroxy group. It is this hydroxy group which is preferably activated. However, it will be understood that the type and amount of the reactive groups may be varied to achieve a covalently conjugated PEG/polypeptide of the present invention.

(103) Water soluble polyoxyethylated polyols are also useful in the present invention. They include polyoxyethylated sorbitol, polyoxyethylated glucose, polyoxyethylated glycerol (POG), etc. POG is preferred. One reason is because the glycerol backbone of polyoxyethylated glycerol is the same backbone occurring naturally in, for example, animals and humans in mono-, di-, triglycerides. Therefore, this branching would not necessarily be seen as a foreign agent in the body. The POG has a preferred molecular weight in the same range as PEG. The structure for POG is shown in Knauf et al., 1988, and a discussion of POG/IL-2 conjugates is found in U.S. Pat. No. 4,766,106.

(104) Another drug delivery system for increasing circulatory half-life is the liposome. The peptides and nucleic acids of the invention may also be administered via liposomes, which serve to target a particular tissue, such as lymphoid tissue, or to target selectively infected cells, as well as to increase the half-life of the peptide and nucleic acids composition. Liposomes include emulsions, foams, micelles, insoluble monolayers, liquid crystals, phospholipid dispersions, lamellar layers and the like. In these preparations, the peptide or nucleic acids to be delivered is incorporated as part of a liposome or embedded, alone or in conjunction with a molecule which binds to a receptor prevalent among lymphoid cells, such as monoclonal antibodies which bind to the CD45 antigen, or with other therapeutic or immunogenic compositions. Thus, liposomes either filled or decorated with a desired peptide or nucleic acids of the invention can be directed to the site of lymphoid cells, where the liposomes then deliver the peptide and nucleic acids compositions. Liposomes for use in accordance with the invention are formed from standard vesicle-forming lipids, which generally include neutral and negatively charged phospholipids and a sterol, such as cholesterol. The selection of lipids is generally guided by consideration of, e.g., liposome size, acid lability and stability of the liposomes in the blood stream. A variety of methods are available for preparing liposomes, as described in, e.g., Szoka et al, 1980, and U.S. Pat. Nos. 4,235,871, 4,501,728, 4,837,028, and 5,019,369.

(105) For targeting cells of the immune system, a ligand to be incorporated into the liposome can include, e.g., antibodies or fragments thereof specific for cell surface determinants of the desired immune system cells. A liposome suspension containing a peptide may be administered intravenously, locally, topically, etc. in a dose which varies according to, inter alia, the manner of administration, the peptide being delivered, and the stage of the disease being treated. For example, liposomes carrying either immunogenic polypeptides are known to elicit CTL responses in vivo (Reddy et al., 1992; Collins et al., 1992; Fries et al., 1992; Nebel et al., 1992).

(106) After the liquid pharmaceutical composition is prepared, it is preferably lyophilized to prevent degradation and to preserve sterility. Methods for lyophilizing liquid compositions are known to those of ordinary skill in the art. Just prior to use, the composition may be reconstituted with a sterile diluent (Ringer's solution, distilled water, or sterile saline, for example) which may include additional ingredients. Upon reconstitution, the composition is preferably administered to subjects using those methods that are known to those skilled in the art.

(107) Another aspect of the present invention relates to conjugates of the isolated peptides or isolated multimeric peptides according to the present invention. Accordingly, the isolated peptides or isolated multimeric peptides according to the present invention may be an amino acid sequence conjugated at any amino acid sidechain or within the amino acid sequence with any chemical moiety, such as any therapeutic agent, such as any immunomodulating compound.

(108) The terms therapeutic agent, such as immunomodulating agent or virus reservoir purging agent as used herein, includes but is not limited to cytokines, such as interferons, monoclonal antibodies, such as ant-PD1 antibodies, cyclophosphamide, Thalidomide, Levamisole, and Lenalidomide.

(109) A virus reservoir purging agent, includes but is not limited to auranofin, IL-7, prostratin, bryostatin, HDAC inhibitors, such as vorinostat, and Disulfuram.

(110) Use of the peptides for evaluating immune responses:

(111) The peptides according to the present invention may be used as diagnostic reagents. For example, a peptide of the invention may be used to determine the susceptibility of a particular individual to a treatment regimen which employs the peptide or related peptides, and thus may be helpful in modifying an existing treatment protocol or in determining a prognosis for an affected individual. In addition, the peptides may also be used to predict which individuals will be at substantial risk for developing a chronic virus infection.

(112) Accordingly, the present invention relates to a method of determining the outcome for a subject exposed to a virus, comprising the steps of determining whether the subject has an immune response to one or more peptides according to the present invention.

(113) In a preferred embodiment of the invention, the peptides as described herein can be used as reagents to evaluate an immune response. The immune response to be evaluated can be induced by using as an immunogen any agent that may result in the production of antigen-specific CTLs or HTLs that recognize and bind to the peptide(s) to be employed as the reagent. The peptide reagent need not be used as the immunogen. Assay systems that can be used for such an analysis include relatively recent technical developments such as tetramers, staining for intracellular lymphokines and interferon release assays, or ELISPOT assays.

(114) For example, a peptide of the invention may be used in a tetramer staining assay to assess peripheral blood mononuclear cells for the presence of antigen-specific CTLs following exposure to an antigen or an immunogen. The HLA-tetrameric complex is used to directly visualize antigen-specific CTLS (see, e.g., Ogg et al., 1998; and Altman et al., 1996) and determine the frequency of the antigen-specific CTL population in a sample of peripheral blood mononuclear cells. A tetramer reagent using a peptide of the invention may be generated as follows: a peptide that binds to an HLA molecule is refolded in the presence of the corresponding HLA heavy chain and beta2-microglobulin to generate a trimolecular complex. The complex is biotinylated at the carboxyl terminal end of the heavy chain at a site that was previously engineered into the protein. Tetramer formation is then induced by the addition of streptavidin. By means of fluorescently labeled streptavidin, the tetramer can be used to stain antigen-specific cells. The cells may then be identified, for example, by flow cytometry. Such an analysis may be used for diagnostic or prognostic purposes. Cells identified by the procedure can also be used for therapeutic purposes. As an alternative to tetramers also pentamers or dimers can be used (Current Protocols in Immunology (2000) unit 17.2 supplement 35)

(115) Peptides of the invention may also be used as reagents to evaluate immune recall responses. (see, e.g., Bertoni et al., 1997 and Perma et al., 1991.). For example, patient PBMC samples from individuals with HCV infection may be analyzed for the presence of antigen-specific CTLs or HTLs using specific peptides. A blood sample containing mononuclear cells may be evaluated by cultivating the PBMCs and stimulating the cells with a peptide of the invention. After an appropriate cultivation period, the expanded cell population may be analyzed, for example, for cytotoxic activity (CTL) or for HTL activity.

(116) The peptides may also be used as reagents to evaluate the efficacy of a vaccine.

(117) PBMCs obtained from a patient vaccinated with an immunogen may be analyzed using, for example, either of the methods described above. The patient is HLA typed, and peptide epitope reagents that recognize the allele-specific molecules present in that patient are selected for the analysis. The immunogenicity of the vaccine is indicated by the presence of epitope-specific CTLs and/or HTLs in the PBMC sample.

(118) The peptides of the invention may also be used to make antibodies, using techniques well known in the art (see, e.g. CURRENT PROTOCOLS IN IMMUNOLOGY, Wiley/Greene, N.Y.; and Antibodies A Laboratory Manual, Harlow and Lane, Cold Spring Harbor Laboratory Press, 1989). Such antibodies include those that recognize a peptide in the context of an HLA molecule, i.e., antibodies that bind to a peptide-MHC complex.

(119) In certain embodiments a first monomeric peptide and the at least one second monomeric peptide are associated via a linker; the linker may comprise any peptide linker, or peptide spacer, such as a glycine, a lysine or an arginine linker/spacer, a polyhistidinyl tag, Protein G, and Protein A but it is also possible to use a bis-maleimide linker/spacer, a disulfide linker, or a polyethylene glycol (PEG) linker. In practice, any linker found useful in peptide chemistry is also useful as a linker according to the present invention. Thus, the invention contemplates the use of simple linear peptides which are conjugated or fused to each other, but also peptide combinations where the individual peptides derived from a natural antigen are linked via non-peptide linkers. Use of multiple linker types are also within the scope of the present invention, and it is e.g. also a part of the invention to utilise linear peptides which include intrachain disulphide linkers.

(120) Particularly interesting peptide combinations of the invention are set forth in the preamble to the examples.

(121) In certain embodiments, at least one of the first and at least one second peptides in the peptide combination comprises an N- or C-terminal modification, such as an amidation, acylation, or acetylation.

(122) Since the peptide combinations are contemplated as vaccine agents or diagnostic agents, they are in certain embodiments coupled to a carrier molecule, such as an immunogenic carrier. The peptides of the peptide combinations may thus be linked to other molecules either as recombinant fusions (e.g. via CLIP technology) or through chemical linkages in an oriented (e.g. using heterobifunctional cross-linkers) or nonoriented fashion. Linking to carrier molecules such as for example diphtheria toxin, latex beads (convenient in diagnostic and prognostic embodiments), and magnetic beads (also convenient in diagnostic and prognostic embodiments), polylysine constructs etc, are all possible according to the invention.

(123) The immunogenic carrier is conveniently selected from carrier proteins such as those conventionally used in the art (e.g. diphtheria or tetanus toxoid, KLH etc.), but it is also possible to use shorter peptides (T-helper epitopes) which can induce T-cell immunity in larger proportions of a population. Details about such T-helper epitopes can e.g. be found in WO 00/20027, which is hereby incorporated by reference hereinall immunologic carriers and promiscuous (i.e. universal) T-helper epitopes discussed therein are useful as immunogenic carriers in the present invention.

(124) In certain embodiments, the carrier is a virus like particle, i.e. a particle sharing properties with virions without being infectious. Such virus-like particles may be provided chemically (e.g. Jennings and Bachmann Ann Rev Pharmacol. Toxicol. 2009. 49:303-26 Immunodrugs: Therapeutic VLP-based vaccines for chronic diseases) or using cloning techniques to generate fusion proteins (e.g. Peabody et al. J. Mol. Biol. 2008; 380: 252-63. Immunogenic display of diverse peptides on virus-like particles of RNA phage MS2). Another example is Remune, an HIV vaccine originally made by Immune Response Corporation, which consists of formalin inactivated HIV that has been irradiated to destroy the viral genome.

(125) In an embodiment, a nucleic acid is encoding one or more monomeric peptide of the multimeric, such as dimeric peptide according to the invention, where the encoded first peptide and the encoded at least one second peptide of a multimeric peptide are associated via a peptide linker, including a peptide spacer, and/or a disulphide bridge. The peptide linker/spacer is typically selected from the group consisting of a glycine, an arginine, a lysine linker/spacer, or a glycine-lysine linker/spacer, but any peptide linker known in the art may be useful. The term peptide linker thus also is intended to denote coupling between the first and second peptide via a peptide bond. A peptide linker that links a first and second peptide by standard peptide bonds may also be referred to as a peptide spacer. Also, the first and second peptides may be linked via a peptide linker and a disulphide bond, as is the case when an intrachain disulphide bond is established.

(126) In one embodiment, the nucleic acid according to the invention encodes the peptide combination, which is coupled (by fusion) to a carrier molecule, such as an immunogenic carrier; useful carriers are discussed above.

(127) In some embodiments the linker is selected from the group consisting of a bis-maleimide linker, a disulfide linker, a polyethylene glycol (PEG) linker, a glycine linker/spacer, a lysine linker/spacer, and an arginine linker/spacer.

(128) In some embodiments the multimeric peptide, such as a dimeric peptide contain a linker in the free amino group of the N-terminal of a monomeric peptide linking said monomeric peptide to another monomeric peptide.

(129) In some embodiments the multimeric peptide, such as a dimeric peptide contain a linker in the free carboxyl group of the C-terminal of a monomeric peptide linking said monomeric peptide to another monomeric peptide.

(130) At least two options for such linkers are described in A. R Jacobson et al, J. Med. Chem. 1989, 32, 1708-1717 and in D Giannotti et al, Journal of Medicinal Chemistry, 2000, Vol. 43, No. 22, the disclosures of which is hereby incorporated by reference.

(131) Alternatively a link between the N-termini of peptides may be established by reacting with Br(CH.sub.2).sub.nBr.

(132) The length of the linker may be varied by the addition of glycine residues, for example Fmoc-NHCH.sub.2CH.sub.2NH-Gly-NH.sub.2 may be used.

(133) An example of such a synthesis, wherein a dimeric peptide is prepared by conjugation through succinic acid, may be as follows:

(134) TABLE-US-00010 (H-Arg-Gly-Thr-Pro-Ile-Har-Gln-Asp-Trp-Gly-Asn- Arg-Ala-Asn-Arg-Gly-Thr-Pro-Thr-Arg-Gln-Glu- Trp-Asp-Cys-Arg-Ile-Ser-NH2Arg-Gly-Thr-Pro-Ile- Har-Gln-Asp-Trp-Gly-Asn-Arg-Ala-Asn-Arg-Gly-Thr- Pro-Thr-Arg-Gln-Glu-Trp-Asp-Cys-Arg-Ile-Ser-NH.sub.2) E(H-Arg-Gly-Thr-Pro-Thr-Har-Asn-Gly-Trp-Asp-Val- Lys-Leu-Ser-Arg-Gly-Thr-Pro-Ile-Har-Gln-Glu-Trp- Har-Ser-Leu-Nle-Asn-Gln-Glu-Trp-NH.sub.2)F (SuccinicacidlinkerbetweenArg.sup.1EandArg.sup.1F)

(135) This dimer was produced from the reaction of the following 2 monomers:

(136) TABLE-US-00011 MonomerE H-Arg-Gly-Thr-Pro-Ile-Har-Gln-Asp-Trp-Gly-Asn-Arg- Ala-Asn-Arg-Gly-Thr-Pro-Thr-Arg-Gln-Glu-Trp-Asp- Cys-Arg-Ile-Ser-NH2Arg-Gly-Thr-Pro-Ile-Har-Gln- Asp-Trp-Gly-Asn-Arg-Ala-Asn-Arg-Gly-Thr-Pro-Thr- Arg-Gln-Glu-Trp-Asp-Cys-Arg-Ile-Ser-NH.sub.2 MonomerF H-Arg-Gly-Thr-Pro-Thr-Har-Asn-Gly-Trp-Asp-Val-Lys- Leu-Ser-Arg-Gly-Thr-Pro-Ile-Har-Gln-Glu-Trp-Har- Ser-Leu-Nle-Asn-Gln-Glu-Trp-NH.sub.2

(137) The two monomers are reacted to give a heterodimer according to the reaction scheme outlined below; where the link is between N-terminal on Arg.sup.1 of on chain E and the N-terminal on Arg.sup.1 in chain F.

(138) Monomers E and F are synthesized separately on a Sieber Amid resin. The Fmoc-groups on N-terminal Gly are removed while the peptides are still on resin. The peptides are cleaved from resin. The resulting protected peptide E is reacted with succinic acid anhydride and thereafter reacted with the protected peptide F. Protective groups are subsequently removed with 95% TFA. The formed heterodimer may be purified from un-reacted monomers by conventional purification methods known to the person skilled in the art.

(139) An example of a synthesis, wherein a dimeric peptide is prepared by conjugation through di-amino propane, may be as follows:

(140) TABLE-US-00012 (H-Gly-Gly-Ala-Lys-Arg-Arg-Val-Val-Gln-Arg-Glu- Lys-Arg-Ala-Gly-Glu-Arg-Glu-Lys-Arg-Ala-Gly-Gly) G(H-Gly-Gly-Ile-Glu-Glu-Glu-Gly-Gly-Arg-Asp-Arg- Asp-Arg-Gly-Gly-Glu-Gln-Asp-Arg-Asp-Arg-Gly-Gly)H trifluoroacetatesalt(Diaminopropanelinker betweenGly.sup.23andGly.sup.23)

(141) This dimer was produced from the reaction of the following 2 protected monomers

(142) TABLE-US-00013 MonomerG H-Arg-Gly-Thr-Pro-Ile-Har-Gln-Asp-Trp-Gly-Asn-Arg- Ala-Asn-Arg-Gly-Thr-Pro-Thr-Arg-Gln-Glu-Trp-Asp- Cys-Arg-Ile-Ser-COOH MonomerH H-Arg-Gly-Thr-Pro-Thr-Har-Asn-Gly-Trp-Asp-Val-Lys- Leu-Ser-Arg-Gly-Thr-Pro-Ile-Har-Gln-Glu-Trp-Har- Ser-Leu-Nle-Asn-Gln-Glu-Trp-COOH

(143) The two monomers G and H are reacted to give a heterodimer according to the reaction scheme outlined below; where the link is between C-terminal on Ser.sup.28 of on chain G and the C-terminal on Trp.sup.31 in chain H.

(144) Monomers G and H are synthesized separately on a 2-chlorotrityl resin. Boc-Gly-OH is coupled to the peptides on the resin before cleaving them of the resin. The resulting peptides are then Boc-protected, alternatively they may me acetylated before being cleaved of the resin. The resulting protected peptide G is reacted with Fmoc-diaminopropane, Fmoc is deprotected and G is coupled to the C-terminal of the protected peptide H via a peptide bond. Protective groups are subsequently removed with 95% TFA. The formed heterodimer may be purified from un-reacted monomers by conventional purification methods known to the person skilled in the art.

(145) Method for Synthesis of Cys-Lys Bridge:

(146) Exemplified with the preparation of BI-155-3 trifluoroacetate salt

(147) TABLE-US-00014 (H-Arg-Gly-Cys(2-oxo-ethyl)-Thr-Pro-Ile-Har-Gln- Asp-Trp-Gly-Asn-Arg-Ala-Asn-Arg-Gly-Thr-Pro-Thr- Arg-Gln-Glu-Trp-Asp-Cys-Arg-Ile-Ser-NH.sub.2)A(H-Arg- Gly-Lys-Thr-Pro-Thr-Har-Asn-Gly-Trp-Asp-Val-Lys- Leu-Ser-Arg-Gly-Thr-Pro-Ile-Har-Gln-Glu-Trp-Har- Ser-Leu-Nle-Asn-Gln-Glu-Trp-NH.sub.2)B trifluoroacetatesalt(Thioetherbond betweenCys(2-oxo-ethyl).sup.3AandLys.sup.3B)

(148) This dimer was produced from the reaction of the following 2 protected monomers

(149) TABLE-US-00015 MonomerA H-Arg-Gly-Cys-Thr-Pro-Ile-Har-Gln-Asp-Trp-Gly- Asn-Arg-Ala-Asn-Arg-Gly-Thr-Pro-Thr-Arg-Gln- Glu-Trp-Asp-Cys-Arg-Ile-Ser-NH.sub.2 MonomerB H-Arg-Gly-Lys(bromoacetyl)-Thr-Pro-Thr-Har-Asn- Gly-Trp-Asp-Val-Lys-Leu-Ser-Arg-Gly-Thr-Pro-Ile- Har-Gln-Glu-Trp-Har-Ser-Leu-Nle-Asn-Gln-Glu-Trp- NH.sub.2

(150) Or with the preparation of BI-155-4 trifluoroacetate salt

(151) TABLE-US-00016 (H-Gly-Ala-Lys-Arg-Arg-Val-Val-Gly-Gly-Cys(2-oxo- ethyl)-Gly-Gly-Ala-Lys-Arg-Arg-Val-Val-Gln-Arg- Glu-Lys-Arg-Ala-Gly-Glu-Arg-Glu-Lys-Arg-Ala-NH.sub.2) A(H-Gly-Lys-Gly-Gly-Ile-Glu-Glu-Glu-Gly-Gly-Arg- Asp-Arg-Asp-Arg-Gly-Gly-Gln-Asp-Arg-Asp-Arg-NH.sub.2)B trifluoroacetatesalt(Thioetherbond betweenCys(2-oxo-ethyl).sup.9AandLys.sup.2B)

(152) This dimer was produced from the reaction of the following 2 protected monomers:

(153) TABLE-US-00017 MonomerA H-Arg-Gly-Cys-Thr-Pro-Ile-Har-Gln-Asp-Trp-Gly- Asn-Arg-Ala-Asn-Arg-Gly-Thr-Pro-Thr-Arg-Gln- Glu-Trp-Asp-Ala-Arg-Ile-Ser-NH.sub.2 MonomerB H-Arg-Gly-Lys(bromoacetyl)-Thr-Pro-Thr-Har-Asn- Gly-Trp-Asp-Val-Lys-Leu-Ser-Arg-Gly-Thr-Pro-Ile- Har-Gln-Glu-Trp-Har-Ser-Leu-Nle-Asn-Gln-Glu-Trp- NH.sub.2

(154) The 2 monomers are reacted to give a heterodimer according to the reaction scheme outlined below; where the link is created between Lys.sup.3 (bromoacetyl) side chain on chain B and Cys in chain A.

(155) At neutral pH and room temperature, bromoacetyl moieties in buffered aqueous solutions are very reactive towards SH-containing moieties, such as the thiol group in cysteine. Thus, if a cysteine is present on the other peptide sequence, the SH will attack the bromoacetyl to form a intermolecular thioether bridge. When the reaction is buffered with a sodium-containing buffer, such as NaHCO.sub.3, the only byproduct of the reaction is NaBr, an innocuous salt.

(156) The formed heterodimer may be purified from un-reacted monomers by conventional purification methods known to the person skilled in the art.

(157) Method for synthesis of oxime bond between two peptide sequences, an intermolecular bond:

(158) Exemplified with the preparation of BI-155 trifluoroacetate salt

(159) TABLE-US-00018 (H-Arg-Gly-Dpr(Ser)-Thr-Pro-Thr-Har-Asn-Gly-Trp- Asp-Val-Lys-Leu-Ser-Arg-Gly-Thr-Pro-Ile-Har-Gln- Glu-Trp-Har-Ser-Leu-Nle-Asn-Gln-Glu-Trp-NH.sub.2)D(H- Arg-Gly-Dpr(Aoa)-Thr-Pro-Ile-Har-Gln-Asp-Trp-Gly- Asn-Arg-Ala-Asn-Arg-Gly-Thr-Pro-Thr-Arg-Gln-Glu- Trp-Asp-Cys-Arg-Ile-Ser-NH.sub.2)C trifluoroacetatesalt(oximeiscreated betweenDpr(Ser).sup.3DandDpr(Aoa).sup.3C)

(160) This dimer is produced from the reaction of the following two monomers:

(161) TABLE-US-00019 MonomerC H-Arg-Gly-Dpr(Aoa)-Thr-Pro-Ile-Har-Gln-Asp-Trp- Gly-Asn-Arg-Ala-Asn-Arg-Gly-Thr-Pro-Thr-Arg- Gln-Glu-Trp-Asp-Cys-Arg-Ile-Ser-NH.sub.2 MonomerD H-Arg-Gly-Dpr(Ser)-Thr-Pro-Thr-Har-Asn-Gly-Trp- Asp-Val-Lys-Leu-Ser-Arg-Gly-Thr-Pro-Ile-Har-Gln- Glu-Trp-Har-Ser-Leu-Nle-Asn-Gln-Glu-Trp-NH.sub.2

(162) The two monomers are reacted to give a heterodimer according to the reaction scheme outlined below; where the link is created between Dpr(Aoa).sup.3 side chain on chain C and oxidized Dpr(Ser) in chain D.

(163) After removal of the Mtt group from Lys and while the peptide was still attached to the resin aminooxyacetylated (AoA) monomer C was synthesized by coupling aminooxyacetic acid to Lys. The peptide was then cleaved from the solid phase support and purified by conventional purification methods. The monomer D was, after cleavage from resin and purification, created by oxidation of the serinyl diaminopropionic acid residue (Dpr(Ser)) with periodate to the aldehyde function. Equimolar amounts of monomer A and B were dissolved in acetonitrile and acetate buffer (pH 4). After reaction for 16 h at room temperature, the product C-oxime-D was isolated by conventional purification methods known to the person skilled in the art.

(164) Dpr=diaminopropionic acid

(165) Fmoc-Dpr (Boc-Ser(tBu))-OH Merck 04-12-1186

(166) Method for Synthesis of Dimers with PEG-Linker:

(167) A multimeric, such as dimeric peptide, such as a heterodimeric peptide may be synthesized by, but are not restricted to the following protocol:

(168) To the peptidyl resin containing deblocked Asp or Glu residue (monomer 1) is added HBTU, DIPEA and Trt-amino PEG amine in DMF. The mixture is allowed to couple over night. The resin is filtered from the solution and washed by standard protocol. The Trt group is removed from the Trt-PEGylated peptide. The monomer 2 containing deblocked Asp or Glu residue is then coupled to the exposed amino group using HBTU and DIPEA. After cleavage the desired product is purified using any suitable technique to give the desired multimeric peptide.

(169) In some embodiments the isolated monomeric peptide contain intramolecular bonds, such as in the form of intramolecular Cys-Cys bonds. It is to be understood that the intramolecular bond, used interchangeably with intrachain bond, is a bond between two different amino acids within the same peptide chain, which however is not necessarily adjacent to each other in the peptide sequence. Accordingly, in some embodiments, the isolated multimeric peptide according to the invention may contain both intramolecular bonds within one or more of the monomers, as well as an intermolecular bond between two chains of the multimeric peptide, such as a dimer. This intramolecular bond may be in the form of Cys-Cys bonds formed with cysteine residues within the same peptide sequence. In some embodiments the monomer contains an intramolecular bond derived from a Lys residue or other amino acid residue, such as a Ser, Cys, Asp or Glu that make the bond, such as a thioether bond or an oxime bond or through a PEG linker, to an amino acid residue on the other monomer peptide sequence.

(170) Method for Synthesis of Multimeric Peptides with PolyLys or MAPS:

(171) PolyLys or MAPS (multiple antigen peptides)has been extensively used over the last 20 years as a carrier protein to produce strong immunogenic response. The MAP system utilizes a peptidyl core of three or more radially branched lysine core to form a backbone for which the epitope sequences of interest can be built parallel using standard solid-phase chemistry.

(172) The MAP system is a commercial product available from several companies such as AnaSpec, Bio-synthesis Inc. and others. The product, as offered in the catalogue only allows attachment of two (identical) peptide sequence to the polyLys core. It is however possible also to link two different peptide sequences by using different protecting groups for alfa- and epsylon-amino functional groups of lysine on the two different peptide sequences.

(173) Use of the MAP system has been described in references including: Wang, C. Y et al. Long-term high-titer neutralizing activity induced by octameric synthetic HIV antigen Science 254, 285-288 (1991). Posnett, D. et al. A novel method for producing anti-peptide antibodies J. Biol. Chem. 263, 1719-1725 (1988), and in Tam, J. P. Synthetic peptide vaccine design: synthesis and properties of a high-density multiple antigenic peptide system PNAS USA 85, 5409-5413 (1988).

(174) The MAP system could also be prepared by chemical (thioether, oxime, hydrazone) ligation of appropriately functionalized tetra- or octavalent polylysine constructs with the peptide antigen. By the use of this chemical ligation, the two peptide sequences being linked together would not have to be identical as they are synthesized separately.

(175) Additionally a novel application of the MAP-based system is to synthesize on solid support a probe containing a poly(ethylene glycol) (PEG) chain in the dendritic arms of MAP.

(176) Use of the MAP system will increase the size of a multimeric complex and may increase the immunogenic response.

(177) Methods for the Synthesis of Multimeric Peptides Using PEG:

(178) Suitable Multi-Arm Activated PEG to be used for a PEG linker are commercially available, e.g. a compound with the following structure:

(179) ##STR00001##

(180) Wherein X may be ethanethiol CH2CH2SH (could be used to form SS bridge with the epitope or a thioether link) or propylamine CH2CH2CH2NH2, among others. These handles preferably allows for the linking of two identical peptide sequences and may be seen as a poly-monomeric epitope presenting construct. One could, however, anchor a dimer (two epitopes linked together) to the PEG above.

(181) Method for Synthesis of Peptide-Poly-L-Lys (PLL)-Polyethylene Glycol (PEG) Construct:

(182) PeptidePLL-PEG constructs, may be synthesized by, but are not restricted to the following protocol:

(183) Fmoc-Poly-L-Lys-resin (a commercial product) is de-protected with 20% piperifine-DMF. Fmoc-NH-PEG.sub.4-COOH, in a mixed solvent of CH.sub.2Cl.sub.2-NMP is added followed by HBTU and DIPEA and the reaction is allowed to proceed for 24 h. The resultant pegylated poly-L-Lys-resin is washed and the pegylation step is repeated. The reaction is monitored by Kaiser's ninhydrin test until a negative reading is obtained. After de-protection of Fmoc group, four identical peptide chains are synthesized directly on the branched poly-L-Lys-polyethylene glycol core by a stepwise solid-phase procedure. All residues activated with HBTU and DIPEA are allowed to couple for 2 h. The coupling is monitored by Kaiser's ninhydrin test and is repeated if needed. After cleavage the desired product is purified using any suitable technique to give the desired peptide-construct.

(184) Table 8 Specific peptides not part of the present invention. (Amino acids underlined refers to place of linker in dimeric molecules; Letter C in a large font refers to a cysteine residue optionally involved in an intramolecular bond with another cysteine residue in the same peptide sequence. Homoarginine is abbreviated Har, Norleucine is abbreviated as Nle or alternatively with the single letter Z, N--methylated Lys is abbreviated Lys(Me), Citrulline is abbreviated with the single letter B, diaminopropionic acid is abbreviated with Dpr and serinyl diaminopropionic acid is abbreviated Dpr(Ser). Flu; abbreviation for Influenza).

(185) Table 8 represent peptides not part of the present invention. These peptides relates to monomeric peptides as well as multimeric peptides comprising two or more of these monomeric peptides, each monomeric peptide independently consisting of not more than 60 amino acids with the following structure
X.sup.1-X.sup.2-X.sup.3-X.sup.4-X.sup.5-X.sup.6(formula III),
wherein X.sup.1, X.sup.3 and optional moiety X.sup.5 independently defines a linear sequence of any 1, 2, 3, 4, or 5 amino acid independently selected from glycine, arginine, norleucine, glutamine, serine, lysine, tryptophan, cysteine, or a derivative thereof; X.sup.2, X.sup.4, and optional moiety X.sup.6 each independently defines a linear sequence of 5-17 amino acids, each having more than 50% sequence identity to a specific natural antigen, said monomeric peptides being covalently joined by one or more intermolecular bond.

(186) TABLE-US-00020 Position withrefer- enceto positions inSEQID NO:200, SEQIDNO: 202,andSEQ Refer- IDNO:203. Anti- ence X2- X4- X6- Pro- Chain gen ID X1 X2 X3 X4 X5 X6 SEQ SEQ SEQ tein Flu BI100_ RR SLLTEVETP GCG VETPIR G TPIRNEWG 2- 7- 9- M2 CGn 10 12 16 at Flu BI100_ RR SLZTDIETP GCG IDTPIR G TPIBQDWG 2- 7- 9- M2 CG 10 12 16 Flu BI100- WWGC TDIET CG IDTPIR G TPIBQDWG 5- 7- 9- M2 CGcyc 9 12 16 Flu BI100- RRG CSLLT C SLLTEVQTPIRN GRR SEWGSRSN 2- 2- 13- M2 Cyc_2 5 13 20 A Flu BI150- RRZC SLLTEVQTPIRN GRR VETPIRN 2- 7- M2 Dimer 13 13 B Flu BI150- WWQC TPIRSEWGCRSN GRR SNDSS G 9- 19- M2 Dimer 20 23 A Flu BI150- WW SLZTDIETP GCG IDTPIR G TPIBQDWG 2- 7- 9- M2 new 10 12 16 B Flu BI150- RR IDTPIR G TPIBQDWG KG SLZTDIETPG 7- 9- 2- M2 new (Har) 12 16 11 A Flu BI150- R SLZTDIETP Dpr IDTPIR G TPIBQDWG 2- 7- 9- M2 2mod 10 12 16 B Flu BI150- RR IDTPIR GG TPI(Har) Dpr SLZTDIETPG 7- 9- 2- M2 2mod QEW (Ser) 12 15 11 A Flu BI150- RR SLZTDIETP GCG IDTPIR G TPIBQDWG 2- 7- 9- M2 dim_2 10 12 16 B Flu BI150- Har IDTPIR G TPIBQDWG KG SLZTDIETPG 7- 9- 2- M2 dim_2 12 16 11 HIV BI450- W.sub.DWGC AKRRV CGG AKRRVVQREKRA 501- 501- gp120 AdjBT1 505 512 HIV BI450- W.sub.DWGC IEEEG CGG IEEEGGERDR 222- 222- gp41 AdjBT2 226 HIV CGG AKRRVV GG AKRRVV G QREKRAV 501- 501- 507- 506 506 513 HIV CGGG DQQLL GG AEEEIV GG IEEEGGERDRDR 257- 266- 221- 261 271 232 HIV CGG AKRRVV GG AKRRVV GG QREKR 501- 501- 507- 506 506 511 HIV CGGG DQQLL GG AEEEIV GG IEEEGG 257- 266- 222- 261 271 227 HIV CGG AEEEVV GG DQQLL 266- 257- 271 261 HIV GCGG AKRRVV GG AKRRVV 501- 501- 506 506 A HIV BI400- G AKRRVV GGCGG AKRRVVQREKRA G EREKRA 501- 501- 507- gp120 B(a- 506 512 512 chain) B HIV BI400- GKG GIEEE GG RDRDR GG EQDRDR 221- 229- 228- gp41 B(b- 225 233 233 chain) E HIV GG AKRRVVQREKRA G EREKRA 501- 507- gp120 512 512 F HIV G GIEEE GG RDRDR GG EQDRDR 221- 229- 228- gp41 225 233 233 G HIV GG AKRRVVQREKRA G EREKRA GG 501- 507- gp120 512 512 H HIV G GIEEE GG RDRDR GG EQDRDRGG 221- 229- 228- gp41 225 233 235 A HIV 400- G AKRRVV GGCGG AKRRVVQREKRA G EREKRA 501- 501- 507- gp120 SeqB 506 512 512 (a- chain) B HIV 400- GKG GIEEE GG RDRDR GG QDRDR 221- 229- 229- gp41 SeqB 225 233 233 (b- chain) D HIV 400- G AKRRVV GG(Dpr AKRRVVQREKRA G EREKRA 501- 501- 507- gp120 SeqB* (Ser)) 506 512 512 (a- GG chain) C HIV 400- GKG GIEEE GG RDRDR GG QDRDR 221- 229- 229- gp41 SeqB* 225 233 233 (b- chain) A HIV BI400- G AKRRVV GGCGG AKRRVVQREKRA G EREKRA 501- 501- 507- gp120 Bu1 506 512 512 (a- chain) B HIV BI400- GKG GIEEE GG ERDRDR GG QDRDR 221- 228- 229- gp41 Bu1 225 233 233 (b- chain) A HIV BI400- G AKRRVV GGCGG AKRRVVEREKRA G QREKRA 501- 501- 507- gp120 Bu2 506 512 512 (a- chain) B HIV BI400- GKG GIEEE GG QDRDR GG RDRDR 221- 229- 229- gp41 Bu2 225 233 233 (b- chain) A HIV BI400- G AKRRVV GGCGG AKRRVVEREKRA G QREKRA 501- 501- 507- gp120 Bu3 506 512 512 (a- chain) B HIV BI400- GKG GIEEE GG EQDRDR GG ERDRD 221- 228- 228- gp41 Bu3 225 233 232 (b- chain) A HIV SEQ400_ GC AKRRVV CGGKG AKRRVVQREKRA G EREKRA 501- 501- 507- gp120 B 506 512 512 (Cyc) B HIV SEQ400_ GKG GIEEE GG RDRDR GG EQDRDR 221- 229- 228- gp41 B 225 233 233 (Cyc) A HIV SEQ400_ GC AKRRVV CGGKG GAKRRVVQREKRA G EREKRA 501- 501- 506- gp120 B 506 512 512 (Cyc) B HIV SEQ400_ GCGG IEEEGGRDRDR GG QDRDR 222- 229- gp41 B 233 233 (Cyc) A HIV BI400- G CAKRRVVC GGKGG AKRRVVQREKRA G EREKRA 501- 501- 507- gp120 bu1 506 512 512 (Cyc) B HIV BI400- CGG IEEEGGERDRDR GG QDRDR 222- 229- gp41 bu1 233 233 (Cyc) A HIV BI400- G CAKRRVVC GGKGG AKRRVVEREKRA G QREKRA 501- 501- 507- gp120 bu2 506 512 512 (Cyc) B HIV BI400- CGG IEEEGGQDRDR GG RDRDR 222- 229- gp41 bu2 233 233 (Cyc) A HIV BI400- G CAKRRVVC GGKGG AKRRVVEREKRA G QREKRA 501- 501- 507- gp120 bu3 506 512 512 (Cyc) B HIV BI400- CGG IEEEGGEQDRDR GG RDRDR 222- 229 gp41 bu3 233 233 (Cyc) A HIV BI400- G CAKRRVVC GGKGG AKRRVVQREKRA G EREKRA 501- 501- 507- gp120 rev 506 512 512 (Cyc) B HIV BI400- CGG EEEIGGRDRD GG RDRDQ 222- 229- gp41 rev 233 233 (Cyc) A HIV BI450- GG RLEPWKH GC GSQPKTA G HPGSQ 7- 15- 13- Tat 1(a- 13 21 17 chain) B HIV BI450- GG FHSQV C FITKGLGISYGRK 32- 38- Tat 1(b- 36 50 chain) A HIV BI450- RLEPWKH GC GSQPKTA GWK HPGSQ 7- 15- 13- Tat 1_2 13 21 17 (a- chain) B HIV BI450- C FITKGLGISY G FITKGLGISYGRK 38- 38- Tat 1_2 47 50 (b- chain) A HCV BI350- RR LLADARVCS GG LLADARVSA 342- 342- E2 1(a- 350 350 chain) B HCV BI350- R GV(Nle) C GVLAGIAYYS 163- 163- E1 1(b- AGIAYFS 172 172 chain) A HCV BI350- RR GNWAKVL K NWAKVI 366- 367- E1 1mod1 372 372 B HCV BI350- RRG LLADARV GCG SGADRV CS 342- 342- E2 1mod1 348 348 A HCV BI350- RR GNWAKVL Dpr NWAKVI 366- 367- E1 1mod2 372 372 B HCV BI350- RRG LLADARV G(Dpr SGADRV CS 342- 342- E2 1mod2 (Ser)) 348 348 G A HCV RR GNWAKVL Lys NWAKVI 366- 367- E1 (Me) 372 372 B HCV RRG LLADARV GEG SGADRV CS 342- 342- E2 348 348 A HCV RR GNWAKVL Lys NWAKVI 366- 367- E1 (Me) 372 372 B HCV RRG LLADARV GDG SGADRV CS 342- 342- E2 348 348 A HCV RR GNWAKVL E NWAKVI 366- 367- E1 372 372 B HCV RRG LLADARV G(Lys SGADRV CS 342- 342- E2 (Me)) 348 348 G A HCV RR GNWAKVL D NWAKVI 366- 367- E1 372 372 B HCV RRG LLADARV G(Lys SGADRV CS 342- 342- E2 (Me)) 348 348 G

SPECIFIC EMBODIMENTS OF THE INVENTION

(187) In some embodiments the isolated peptide according to the present invention has a total of not more than 60 amino acids.

(188) In some embodiments the sequence of amino acids defined by (Z.sup.1-Z.sup.2).sub.1-Z.sup.3-(Z.sup.4-Z.sup.5).sub.2-Z.sup.6-(Z.sup.7-Z.sup.8).sub.3-Z.sup.9-(Z.sup.10-Z.sup.11).sub.4-Z.sup.12 is not found in any native sequence of a protein.

(189) In some embodiments the peptide according to the present invention is demonstrated to translocate across a plasma membrane in the assay based on biotinylation of peptides as described in example 5.

(190) In some embodiments Z.sup.3, and optional Z.sup.6, Z.sup.9 and Z.sup.12 defines an amino acid sequence identical to the native sequence of a known antigen.

(191) In some embodiments Z.sup.3, and optional Z.sup.6, Z.sup.9 and Z.sup.12 defines an amino acid sequence not identical to the native sequence of any known antigen.

(192) In some embodiments Z.sup.3, and optional Z.sup.6, Z.sup.9 and Z.sup.12 defines any chemical moiety, which is any therapeutical compound, such as an immunomodulating compound, such as a Cox-2 inhibitor.

(193) In some embodiments the peptide according to the present invention is capable of inducing a T-lymphocyte response.

(194) In some embodiments the peptide according to the present invention is capable of inducing a CD4+ and/or a CD8+ T-lymphocyte response.

(195) In some embodiments the antigen is a viral protein, such as a capsid protein.

(196) In some embodiments the viral protein is selected from a protein of the Hepatitis C virus, such as a core protein; protein of influenza virus, such as an M2 protein.

(197) In some embodiments the viral protein of Hepatitis C virus is selected from HCV consensus sequence of genotype 1, such as subtypes 1a and 1b, genotype 2 such as 2a and 2b and genotype 3, such as 3a.

(198) In some embodiments, in the peptide according to the present invention, the specific natural antigen is a protein or peptide sequence derived from a disease antigen, such as an infectious agent, such as bacteria, virus, parasite, fungus, or cancer antigens such as oncogene (lung, stomach, breast cancer) or an antigen causing an autoimmune disease such as diabetes, multiple sclerosis (MS), celiac disease, Myalgic Encephalomyelitis (ME), psoriasis, and/or Crohn's Disease.

(199) Accordingly confirmed and suspected autoimmune diseases, where relevant antigens may be derived include Achlorhydra Autoimmune Active Chronic Hepatitis, Acute Disseminated Encephalomyelitis, Acute hemorrhagic leukoencephalitis, Addison's Disease, Agammaglobulinemia, Alopecia greata, Amyotrophic Lateral Sclerosis, Ankylosing Spondylitis, Anti-GBM/TBM Nephritis, Antiphospholipid syndrome, Antisynthetase syndrome, Arthritis, Atopic allergy, Atopic Dermatitis, Autoimmune Aplastic Anemia, Autoimmune cardiomyopathy, Autoimmune hemolytic anemia, Autoimmune hepatitis, Autoimmune inner ear disease, Autoimmune lymphoproliferative syndrome, Autoimmune peripheral neuropathy, Autoimmune pancreatitis, Autoimmune polyendocrine syndrome Types I, II, & III, Autoimmune progesterone dermatitis, Autoimmune thrombocytopenic purpura, Autoimmune uveitis, Balo disease/Balo concentric sclerosis, Bechets Syndrome, Berger's disease, Bickerstaff's encephalitis, Blau syndrome, Bullous Pemphigoid, Castleman's disease, Chagas disease, Chronic Fatigue Immune Dysfunction Syndrome, Chronic inflammatory demyelinating polyneuropathy, Chronic recurrent multifocal ostomyelitis, Chronic lyme disease, Chronic obstructive pulmonary disease, Churg-Strauss syndrome, Cicatricial Pemphigoid, Coeliac Disease, Cogan syndrome, Cold agglutinin disease, Complement component 2 deficiency, Cranial arteritis, CREST syndrome, Crohns Disease (one of two types of idiopathic inflammatory bowel disease IBD), Cushing's Syndrome, Cutaneous leukocytoclastic angiitis, Dego's disease, Dercum's disease, Dermatitis herpetiformis, Dermatomyositis, Diabetes mellitus type 1, Diffuse cutaneous systemic sclerosis, Dressler's syndrome, Discoid lupus erythematosus, Eczema, Endometriosis, Enthesitis-related arthritis, Eosinophilic fasciitis, Epidermolysis bullosa acquisita, Erythema nodosum, Essential mixed cryoglobulinemia, Evan's syndrome, Fibrodysplasia ossificans progressiva, Fibromyalgia, Fibromyositis, Fibrosing aveolitis, Gastritis, Gastrointestinal pemphigoid, Giant cell arteritis, Glomerulonephritis, Goodpasture's syndrome, Graves' disease, Guillain-Barr syndrome (GBS), Hashimoto's encephalitis, Hashimoto's thyroiditis, Haemolytic anaemia, Henoch-Schonlein purpura, Herpes gestationis, Hidradenitis suppurativa, Hughes syndrome (See Antiphospholipid syndrome), Hypogammaglobulinemia, Idiopathic Inflammatory Demyelinating Diseases, Idiopathic pulmonary fibrosis, Idiopathic thrombocytopenic purpura (See Autoimmune thrombocytopenic purpura), IgA nephropathy (Also Berger's disease), Inclusion body myositis, Inflammatory demyelinating polyneuopathy, Interstitial cystitis, Irritable Bowel Syndrome (IBS), Juvenile idiopathic arthritis, Juvenile rheumatoid arthritis, Kawasaki's Disease, Lambert-Eaton myasthenic syndrome, Leukocytoclastic vasculitis, Lichen planus, Lichen sclerosus, Linear IgA disease (LAD), Lou Gehrig's Disease (Also Amyotrophic lateral sclerosis), Lupoid hepatitis, Lupus erythematosus, Majeed syndrome, Mnire's disease, Microscopic polyangiitis, Miller-Fisher syndrome, Mixed Connective Tissue Disease, Morphea, Mucha-Habermann disease, Muckle-Wells syndrome, Multiple Myeloma, Multiple Sclerosis, Myasthenia gravis, Myositis, Narcolepsy, Neuromyelitis optica (Also Devic's Disease), Neuromyotonia, Occular cicatricial pemphigoid, Opsoclonus myoclonus syndrome, Ord thyroiditis, Palindromic rheumatism, PANDAS (Pediatric Autoimmune Neuropsychiatric Disorders Associated with Streptococcus), Paraneoplastic cerebellar degeneration, Paroxysmal nocturnal hemoglobinuria (PNH), Parry Romberg syndrome, Parsonnage-Turner syndrome, Pars planitis, Pemphigus, Pemphigus vulgaris, Pernicious anaemia, Perivenous encephalomyelitis, POEMS syndrome, Polyarteritis nodosa, Polymyalgia rheumatica, Polymyositis, Primary biliary cirrhosis, Primary sclerosing cholangitis, Progressive inflammatory neuropathy, Psoriasis, Psoriatic Arthritis, Pyoderma gangrenosum, Pure red cell aplasia, Rasmussen's encephalitis, Raynaud phenomenon, Relapsing polychondritis, Reiter's syndrome, Restless leg syndrome, Retroperitoneal fibrosis, Rheumatoid arthritis, Rheumatoid fever, Sarcoidosis, Schizophrenia, Schmidt syndrome, Schnitzler syndrome, Scleritis, Scleroderma, Sjgren's syndrome, Spondyloarthropathy, Sticky blood syndrome, Still's Disease, Stiff person syndrome, Subacute bacterial endocarditis (SBE), Susac's syndrome, Sweet syndrome, Sydenham Chorea, Sympathetic ophthalmia, Takayasu's arteritis, Temporal arteritis (also known as giant cell arteritis), Tolosa-Hunt syndrome, Transverse Myelitis, Ulcerative Colitis (one of two types of idiopathic inflammatory bowel disease IBD), Undifferentiated connective tissue disease, Undifferentiated spondyloarthropathy, Vasculitis, Vitiligo, Wegener's granulomatosis, Wilson's syndrome, and Wiskott-Aldrich syndrome.

(200) In some embodiments, in the peptide according to the present invention, the specific natural antigen is a viral protein, such as a structural protein, such as a capsid protein, a regulatory protein, an enzymatic protein, and a proteolytic protein.

(201) In some embodiments, in the peptide according to the present invention, the viral protein is a protein, such as a structural protein, such as a core or envelope protein, of a virus selected from the Hepatitis C virus; influenza virus such as an M2 protein, human immunodeficiency virus (HIV), cytomegalovirus (CMV), and Human papillomavirus (HPV).

(202) In some embodiments, in the peptide according to the present invention, the viral protein is a viral protein of Hepatitis C virus selected from any one HCV consensus sequence of a specific genotype, such as 1, such as subtypes is and 1b, genotype 2, such as 2a and 2b, genotype 3, such as 3a, genotype 4, genotype 5, and genotype 6.

(203) In some embodiments the peptide according to the present invention is of 19-60 amino acids, such as of 20-60 amino acids, such as of 21-60 amino acids, such as of 22-60 amino acids, such as of 23-60 amino acids, such as of 24-60 amino acids, such as of 25-60 amino acids, such as of 26-60 amino acids, such as of 27-60 amino acids, such as of 28-60 amino acids, such as of 29-60 amino acids, such as of 30-60 amino acids, such as of 31-60 amino acids, such as of 32-60 amino acids, such as of 33-60 amino acids, such as of 34-60 amino acids, such as of 35-60 amino acids.

(204) In some embodiments the peptide according to the present invention is of 18-60 amino acids, such as 18-59 amino acids, such as 18-58 amino acids, such as 18-57 amino acids, such as 18-56 amino acids, such as 18-55 amino acids, such as 18-54 amino acids, such as 18-53 amino acids, such as 18-52 amino acids, such as 18-51 amino acids, such as 18-50 amino acids, such as 18-49 amino acids, such as 18-48 amino acids, such as 18-47 amino acids, such as 18-46 amino acids, such as 18-45 amino acids, such as 18-44 amino acids, such as 18-43 amino acids, such as 18-42 amino acids, such as 18-41 amino acids, such as 18-40 amino acids, such as 18-39 amino acids, such as 18-38 amino acids, such as 18-37 amino acids, such as 18-35 amino acids, such as of 18-34 amino acids, such as of 18-33 amino acids, such as of 18-32 amino acids, such as of 18-31 amino acids, such as of 18-30 amino acids, such as of 18-29 amino acids, such as of 18-28 amino acids, such as of 18-27 amino acids, such as of 18-26 amino acids, such as of 18-25 amino acids, such as of 18-24 amino acids, such as of 18-23 amino acids, such as of 18-22 amino acids, such as of 18-21 amino acids, such as of 18-20 amino acids, such as of 18-19 amino acids.

(205) In some embodiments in the peptide according to the present invention, the monomeric peptide contain one or more intramolecular bond, such as one or more Cys-Cys bond.

(206) In some embodiments in the peptide according to the present invention, the monomeric peptide has delayed proteolytic degradation in the N-terminal, such as by incorporation of the first 1, 2, or 3 amino acids in the N-terminal in the D-form, or by incorporation of the first 1, 2, or 3 amino acids in the N-terminal in beta or gamma form.

(207) In some embodiments, in the multimeric, such as a dimeric peptide according to the present invention, the two or more monomeric peptides are identical in sequence.

(208) In some embodiments, in the multimeric, such as dimeric peptide according to the present invention, the two or more monomeric peptides are different in sequence.

(209) In some embodiments, in the multimeric, such as dimeric peptide according to the present invention, one, two or more of the peptide strands of the multimeric, such as dimeric peptide has delayed proteolytic degradation in the N-terminal, such as by incorporation of the first 1, 2, or 3 amino acids in the N-terminal in the D-form, or by incorporation of the first 1, 2, or 3 amino acids in the N-terminal in beta or gamma form.

(210) In some embodiments, in the multimeric, such as dimeric peptide according to the present invention, the linker is placed within any sequence selected from Z.sup.1, Z.sup.2, Z.sup.3, Z.sup.4, Z.sup.5, Z.sup.6, Z.sup.7, Z.sup.8, Z.sup.9, Z.sup.10, Z.sup.11, and Z.sup.12, such as in Z.sup.1, Z.sup.2, Z.sup.3, Z.sup.4, Z.sup.5, Z.sup.6, Z.sup.7, Z.sup.8, Z.sup.9, Z.sup.10, Z.sup.11, and Z.sup.12 of the first monomeric peptide to anywhere on the at least one second monomeric peptide, such as within the sequence of Z.sup.1, Z.sup.2, Z.sup.3, Z.sup.4, Z.sup.5, Z.sup.6, Z.sup.7, Z.sup.8, Z.sup.9, Z.sup.10, Z.sup.11, and Z.sup.12.

(211) In some embodiments, in the multimeric, such as dimeric peptide according to the present invention, the linker is placed at an amino acid position selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60 of the first monomeric peptide to a position selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60 of the at least one second monomeric peptide.

(212) In some embodiments, in the multimeric, such as dimeric peptide according to the present invention, the multimeric, such as dimeric peptide contain a helper epitope of at least 12 amino acids, such as at least 13, 14, 15 or 17 amino acids, which helper epitope consist of a combined sequence of amino acids, which is a sequence of amino acids from a first specific continuous antigenic peptide sequences, and a sequence of amino acids from at least one second specific continuous antigenic peptide sequence of the same or different protein derived from the same virus, any different virus, or any disease antigen, such as between 2-12 amino acids from the first specific continuous antigenic peptide sequences and 2-12 amino acids from the at least one second specific continuous antigenic antigenic peptide sequence.

(213) In some embodiments, in the isolated peptide according to the present invention, the peptide contain a helper epitope of at least 12 amino acids, such as at least 13, 14, 15 or 17 amino acids, which helper epitope consist of a combined sequence of amino acids, which is a sequence of amino acids from a first specific continuous antigenic peptide sequences, and a sequence of amino acids from at least one second specific continuous antigenic peptide sequence of the same or different protein derived from the same virus, any different virus, or any disease antigen, such as between 2-12 amino acids from the first specific continuous antigenic peptide sequences and 2-12 amino acids from the at least one second specific continuous antigenic antigenic peptide sequence.

(214) It is to be understood that an epitope may not only be present within the sequence of the monomeric peptide. An epitope may also be present with a combination of amino acids of the first and the at least one second monomeric peptide in a multimeric, such as dimeric peptide sequence, wherein this combination of amino acids forms a sequence that span from the first to the at least one second monomeric peptide sequence. This epitope may be a continuous sequence of amino acids or it may be a three-dimensional epitope with amino acids found in both monomeric peptides.

(215) In some embodiments, in the multimeric, such as dimeric peptide according to the present invention, the intermolecular bond is a disulfide (SS) bond between two Cys residues.

(216) In some embodiments, in the multimeric, such as dimeric peptide according to the present invention, the intermolecular bond is a methylated peptide bond between a N--methylated Lys side-chain and the side-chain of an Asp or Glu residue.

(217) In some embodiments, in the multimeric, such as dimeric peptide according to the present invention, the intermolecular bond is a thioether bond between a Cys residue in the first monomeric peptide and a modified Lys residue in the at least one second monomeric peptide.

(218) In some embodiments, in the multimeric, such as dimeric peptide according to the present invention, the intermolecular bond is an oxime bond.

(219) In some embodiments, in the multimeric, such as dimeric peptide according to the present invention, the intermolecular bond is an oxime bond between a derivatized Lys residue in the first monomeric peptide and a derivatized Ser residue in the at least one second monomeric peptide.

(220) In some embodiments, in the multimeric, such as dimeric peptide according to the present invention, the intermolecular bond is an oxime bond between a derivatized lysine, ornitine or diaminopropionic acid residue in the first monomeric peptide and a derivatized serine moiety, such as a serine residue, such as in a serinyl diaminopropionic acid residue, such as in a serinyl lysin residue or such as in a serinyl ornitine residue, in the at least one second monomeric peptide.

(221) In some embodiments, in the multimeric, such as dimeric peptide according to the present invention, the monomeric peptides are linked by a polyethylene glycol (PEG) linker, such as through an Asp or a Glu residue in the first monomeric peptide and an Asp or a Glu residue in the at least one second monomeric peptide.

(222) In some embodiments, in the multimeric, such as dimeric peptide according to the present invention, any one of the monomeric peptides is independently as defined herein.

(223) In some embodiments, the peptide according to the present invention is essentially a non-cell-penetrating peptide. In other embodiments, the peptide according to the present invention is a cell-penetrating peptide. In some embodiments, the peptide according to the present invention is able to attach to the cell membrane of an antigen presenting cell.

(224) It is to be understood that when referring to the peptides ability to attach to and enter a cell, such as an antigen presenting cell, it may be with reference to the complete sequence of the peptide as well as a fragment thereof, such as a fragment representing an epitope. Accordingly, it may be the case that the entire sequence is essentially a non-cell-penetrating peptide, whereas a fragment of the peptide is able to efficiently enter a cell, such as an antigen presenting cell.

(225) In some embodiments, the peptide according to the present invention is not a peptide or a dimeric peptide as specifically disclosed in International Patent Application No: PCT/DK2011/050460.

(226) In some embodiments, the peptide according to the present invention is not a peptide or a dimeric peptide as specifically disclosed in International Patent Application No: PCT/EP2010/059513, such as one selected from:

(227) TABLE-US-00021 (SEQIDNO:267) CGGAKRRVVGGAKRRVVGQREKRAV (SEQIDNO:268) CGGGDQQLLGGAEEEIVGGIEEEGGERDRDR (SEQIDNO:269) CGGAKRRVVGGAKRRVVGGQREKR (SEQIDNO:270) CGGGDQQLLGGAEEEIVGGIEEEGG (SEQIDNO:271) CGGAEEEVVGGDQQLL (SEQIDNO:272) GCGGAKRRVVGGAKRRVV (SEQIDNO:273) GAKRRVVGGCGGAKRRVVQREKRAGEREKRA (SEQIDNO:274) GKGGIEEEGGRDRDRGGEQDRDR (SEQIDNO:275) GAKRRVVGGCGGAKRRVVQREKRAGEREKRA (SEQIDNO:276) GKGGIEEEGGERDRDRGGQDRDR (SEQIDNO:277) GAKRRVVGGCGGAKRRVVEREKRAGQREKRA (SEQIDNO:278) GKGGIEEEGGQDRDRGGRDRDR (SEQIDNO:279) GAKRRVVGGCGGAKRRVVEREKRAGQREKRA (SEQIDNO:280) GKGGIEEEGGEQDRDRGGERDRD

(228) In some embodiments, the peptide according to the present invention is not a dimeric peptide selected from (The peptides are linked via the underlined amino acid):

(229) ##STR00002##

(230) In some embodiments Z.sup.3, and optional Z.sup.6, Z.sup.9 and Z.sup.12 consist of a sequence selected from GYIPLVGAPLG, GYLPAVGAPIG, GYLPAVGAPI, NYVTGNIPG, NYATGNLPG, NYATGNLPG, VTGNIPGSTYS, IRNLGRVIETLTG, SRNLGKVIDTLTC, IRNLGRVIETLT, GGGQIIGGNYLIP, GGGQIVGGVYLLP, LIFLARSALIV, LIFLARSALIL, LIFLARSALIL, SAYERMCNIL, SAYERNleVNIL, TAYERNleCNIL, IAYERMCNIL, IAYERMCNIL, LFFKClYRLFKHGL, LFFKTITRLFBHGL, GLEPLVIAGILA, GSDPLVVAASIV, NLVPMVATV, NLVPMVATV, NIVPNleVVTA, PEVIPMFSALS, FIIPXFTALSG, ALGPAATL, GPVVHLTL, LECVYCKQQLL, GVYDFAFRDLC, GVFDYAFRDIN, GATPVDLLGA, GVTPAGLIGV, VARALAHGVRV, VIRVIAHGLRL, GITFSIFLIVS, CSFSIFLLAL, GCSFSIFLLAL, GITFSIYLIVS, LNleGYIPLIGA, LMGYIPLVGA, LNleGYIPLIGA, PBIGVRATB, GPRLGVRATR, GPRLGVRAT, RGSVAHKS, SALILRGSVAHK, FQTAAQRAMM, FQTAAQRAVNle, FQTVVQBA, FQTAAQRA, GPSTEGVPESM, LLSTEGVPNSNle, GSLVGLLHIVL, ASIVGILHLIL, NLVPMVATV, NIVPNleVVTA, TPQDLNTMLN, ALLYGATPYAIG, MMTACQGVG, GQAGDDFS, EVYDFAFRDLC, GFAFRDLCIVY, GFAYRDINLAY, GALNLCLPM, and GALQIBLPL, IRNLGRVIETLTLNleGYIPLIGA, or a fragment or variant thereof.

(231) In some embodiments Z.sup.3, and optional Z.sup.6, Z.sup.9 and Z.sup.12 consist of a sequence derived from an amino acid sequence selected from SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:126, SEQ ID NO:198, SEQ ID NO:198, SEQ ID NO:200, SEQ ID NO:201, SEQ ID NO:202, SEQ ID NO:203, SEQ ID NO:204, and SEQ ID NO:205, or a fragment or variant thereof.

(232) In some embodiments the peptide according to the invention is not a peptide selected from RRGYIPLVGAPLGBGRVARALAHGVRV, RGYIPLVGAPLGRRVARALAHGVRV, RGYIPLVGAPLGRRRVARALAHGVRVR, RRGYIPLVGAPLGRRVARALAHGVRV, RRGYIPLVGAPLGRRRVARALAHGVRV, BRGYIPLVGAPLGRRVARALAHGVRV, RRRGYIPLVGAPLGBRVARALAHGVRV, RGYIPLVGAPLGKKKVARALAHGVRV, RGYIPLVGAPLGRRRVARALAHGVRV, KKGYIPLVGAPLGKKVARALAHGVRV, WGYIPLVGAPLGRRVARALAHGVRV, WWGYIPLVGAPLGRRVARALAHGVRV, EEGYIPLVGAPLGEEVARALAHGVRV, GGGYIPLVGAPLGGGVARALAHGVRV, EEGYIPLVGAPLGRRVARALAHGVRV, RRGYIPLVGAPLGLRRVARALAHGVRV, WWGYIPLVGAPLGRRVARALAHGVRV, WWGYIPLVGAPLGRRRVARALAHGVRV, WWGYIPLVGAPLGRVARALAHGVRV, RGYIPLVGAPLGRRVARALAHGVRV, RRGYLPAVGAPIGBRVIRVIAHGLRL, RRGYIPLVGAPLGBRVARALAHGVRV, GYIPLVGAPLGGVARALAHGVRV, WWGYLPAVGAPIRRVIRVIAHGLRL, GYIPLVGAPLGGVARALAHGVRV, RRGYIPLVGAPLGBGRVARALAHGVRV, RGYIPLVGAPLGRRVARALAHGVRV, RGYIPLVGAPLGRRRVARALAHGVRV, RRGYIPLVGAPLGRRVARALAHGVRV, RRGYIPLVGAPLGRRRVARALAHGVRV, BRGYIPLVGAPLGRRVARALAHGVRV, RRRGYIPLVGAPLGBRVARALAHGVRV, RGYIPLVGAPLGKKKVARALAHGVRV, RGYIPLVGAPLGRRRVARALAHGVRV, KKGYIPLVGAPLGKKVARALAHGVRV, WGYIPLVGAPLGRRVARALAHGVRV, WWGYIPLVGAPLGRRVARALAHGVRV, RRGYIPLVGAPLGLRRVARALAHGVRV, RRNYVTGNIPGBRGITFSIFLIVS, WWNYATGNLPGRRCSFSIFLLAL, WWNYVTGNIPGBRGITFSIFLIVS, WWNYVTGNIPGRRGITFSIFLIVS, RRNYATGNLPGRRGCSFSIFLLAL, RRVTGNIPGSTYSGBRGITFSIYLIVS, RRIRNLGRVIETLTGBRLNleGYIPLIGA, RRSRNLGKVIDTLTCBRLMGYIPLVGA, SRNLGKVIDTLTCGFADLMGYIPLVGA, WWIRNLGRVIETLTRRLNleGYIPLIGA, WWSRNLGKVIDTLTCRRLMGYIPLVGA, RRGGGQIIGGNYLIPRBPBIGVRATB, GGGQIVGGVYLLPRRGPRLGVRATR, RRGGGQIVGGVYLLPRRGPRLGVRATR, WWGGGQIVGGVYLLPRRGPRLGVRAT, BRLIFLARSALIVRGSVAHKS, EDLIFLARSALILRGSVAHKS, BRLIFLARSALILBGRSALILRGSVAHK, SAYERMCNILKGKFQTAAQRAMM, SAYERNleVNILKGKFQTAAQRAVNle, BRTAYERNleCNILBRGRFQTVVQBA, BRIAYERMCNILLBRGKFQTAAQRA, IAYERMCNILKGKFQTAAQRA, LFFKClYRLFKHGLKRGPSTEGVPESM, BRRLFFKTITRLFBHGLRRLLSTEGVPNSNle, BRGLEPLVIAGILARRGSLVGLLHIVL, BRGSDPLVVAASIVRRASIVGILHLIL, RNLVPMVATVRRNLVPMVATVB, RNLVPMVATVBRRNLVPMVATVB, RNIVPNleVVTARRNIVPNleVVTAB, PEVIPMFSALSEGATPQDLNTMLN, RFIIPXFTALSGGRRALLYGATPYAIG, KALGPAATLEEMMTACQGVG, RRGPVVHLTLRRRGQAGDDFS, RRGPVVHLTLRRRGQAGDDFS, RRGPVVHLTLRGRRGQAGDDFS, RRLECVYCKQQLLRREVYDFAFRDLC, RRGVYDFAFRDLCRRGFAFRDLCIVYR, RRGVFDYAFRDINRRGFAYRDINLAYR, RRGATPVDLLGARRGALNLCLPMR, RRGVTPAGLIGVRRGALQIBLPLR, RGYLPAVGAPIGRRRVIRVIAHGLRLR, RRSRNLGKVIDTLTCRRLMGYIPLVGA, RRIRNLGRVIETLTLNleGYIPLIGARRIRNLGRVIETLTLNleGYIPLIGAR, or a fragment or variant thereof.

(233) In some embodiments the peptide according to the invention is not a peptide consisting of a sequence selected from X.sup.1-NYVTGNIPG-X.sup.3-GITFSIYLIVS; X.sup.1-IRNLGRVIETLT-X.sup.3-LNleGYIPLIGA; X.sup.1-GYLPAVGAP.sup.1-X.sup.3-VIRVIAHGLRL; X.sup.1-GGGQIIGGNYLIP-X.sup.3-PBIGVRATB; X.sup.1-NYATGNLPG-X.sup.3-GCSFSIFLLAL; X.sup.1-SRNLGKVIDTLTC-X.sup.3-LMGYIPLVGA; X.sup.1-GYIPLVGAPL-X.sup.3-VARALAHGVRV; X.sup.1-GGGQIVGGVYLLP-X.sup.3-PRLGVRATR; X.sup.1-LTFLVRSVLL.sup.1-X.sup.3-GSVLIVRGSLVH; X.sup.1-TAYERNleCNIL-X.sup.3-GRFQTVVQBA; X.sup.1-SDPLVVAASIV-X.sup.3-ASIVGILHLIL; X.sup.1-LIFLARSALIL-X.sup.3-SALILRGSVAH; X.sup.1-IAYERMCNIL-X.sup.3-GKFQTAAQRA; and X.sup.1-LEPLVIAGILA-X.sup.3-GSLVGLLHIVL; X.sup.1-NLVPMVATV-X.sup.3-NLVPMATV; X.sup.1-GYLPAVGAPIG-X.sup.3-VIRVIAHGLRL; X.sup.1-IRNLGRVIETLTG-X.sup.3-LNleGYIPLIGA; X.sup.1-GVYDFAFRDLC-X.sup.3-GFAFRDLCIVYR, X.sup.1-GVFDYAFRDIN-X.sup.3-GFAYRDINLAYR, X.sup.1-GATPVDLLGA-X.sup.3-GALNLCLPMR, X.sup.1-GVTPAGLIGV-X.sup.3-GALQIBLPLR, and X.sup.1-IRNLGRVIETLTLNleGYIPLIGA-X.sup.3-IRNLGRVIETLTLNleGYIPLIGA; optionally with an X.sup.5 in the C-terminal of the peptide wherein X.sup.1, X.sup.3 and X.sup.5 refers to X.sup.1, X.sup.3, and X.sup.5 of formula II.

(234) In some embodiments the peptide according to the invention is not a peptide consisting of a sequence selected from RRGYIPLVGAPLGBGRVARALAHGVRV (SEQ ID NO:47), RGYIPLVGAPLGRRVARALAHGVRV (SEQ ID NO:48), RGYIPLVGAPLGRRRVARALAHGVRVR (SEQ ID NO:49), RRGYIPLVGAPLGRRVARALAHGVRV (SEQ ID NO:50), RRGYIPLVGAPLGRRRVARALAHGVRV (SEQ ID NO:51), BRGYIPLVGAPLGRRVARALAHGVRV (SEQ ID NO:52), RRRGYIPLVGAPLGBRVARALAHGVRV (SEQ ID NO:53), RGYIPLVGAPLGKKKVARALAHGVRV (SEQ ID NO:54), RGYIPLVGAPLGRRRVARALAHGVRV (SEQ ID NO:55), KKGYIPLVGAPLGKKVARALAHGVRV (SEQ ID NO:56), WGYIPLVGAPLGRRVARALAHGVRV (SEQ ID NO:57), WWGYIPLVGAPLGRRVARALAHGVRV (SEQ ID NO:58), EEGYIPLVGAPLGEEVARALAHGVRV (SEQ ID NO:59), GGGYIPLVGAPLGGGVARALAHGVRV (SEQ ID NO:60), EEGYIPLVGAPLGRRVARALAHGVRV (SEQ ID NO:61), RRGYIPLVGAPLGLRRVARALAHGVRV (SEQ ID NO:62), WWGYIPLVGAPLGRRVARALAHGVRV (SEQ ID NO:63), WWGYIPLVGAPLGRRRVARALAHGVRV (SEQ ID NO:64), WWGYIPLVGAPLGRVARALAHGVRV (SEQ ID NO:65), RGYIPLVGAPLGRRVARALAHGVRV (SEQ ID NO:66), RRGYLPAVGAPIGBRVIRVIAHGLRL (SEQ ID NO:67), RRGYIPLVGAPLGBRVARALAHGVRV (SEQ ID NO:68), GYIPLVGAPLGGVARALAHGVRV (SEQ ID NO:69), WWGYLPAVGAPIRRVIRVIAHGLRL (SEQ ID NO:70), GYIPLVGAPLGGVARALAHGVRV (SEQ ID NO:71), RRGYIPLVGAPLGBGRVARALAHGVRV (SEQ ID NO:72), RGYIPLVGAPLGRRVARALAHGVRV (SEQ ID NO:73), RGYIPLVGAPLGRRRVARALAHGVRV (SEQ ID NO:74), RRGYIPLVGAPLGRRVARALAHGVRV (SEQ ID NO:75), RRGYIPLVGAPLGRRRVARALAHGVRV (SEQ ID NO:76), BRGYIPLVGAPLGRRVARALAHGVRV (SEQ ID NO:77), RRRGYIPLVGAPLGBRVARALAHGVRV (SEQ ID NO:78), RGYIPLVGAPLGKKKVARALAHGVRV (SEQ ID NO:79), RGYIPLVGAPLGRRRVARALAHGVRV (SEQ ID NO:80), KKGYIPLVGAPLGKKVARALAHGVRV (SEQ ID NO:81), WGYIPLVGAPLGRRVARALAHGVRV (SEQ ID NO:82), WWGYIPLVGAPLGRRVARALAHGVRV (SEQ ID NO:83), RRGYIPLVGAPLGLRRVARALAHGVRV (SEQ ID NO:84), RRNYVTGNIPGBRGITFSIFLIVS (SEQ ID NO:85), WWNYATGNLPGRRCSFSIFLLAL (SEQ ID NO:86), WWNYVTGNIPGBRGITFSIFLIVS (SEQ ID NO:87), WWNYVTGNIPGRRGITFSIFLIVS (SEQ ID NO:88), RRNYATGNLPGRRGCSFSIFLLAL (SEQ ID NO:89), RRVTGNIPGSTYSGBRGITFSIYLIVS (SEQ ID NO:90), RRIRNLGRVIETLTGBRLNleGYIPLIGA (SEQ ID NO:91), RRSRNLGKVIDTLTCBRLMGYIPLVGA (SEQ ID NO:92), SRNLGKVIDTLTCGFADLMGYIPLVGA (SEQ ID NO:93), WWIRNLGRVIETLTRRLNleGYIPLIGA (SEQ ID NO:94), WWSRNLGKVIDTLTCRRLMGYIPLVGA (SEQ ID NO:95), RRGGGQIIGGNYLIPRBPBIGVRATB (SEQ ID NO:96), GGGQIVGGVYLLPRRGPRLGVRATR (SEQ ID NO:97), RRGGGQIVGGVYLLPRRGPRLGVRATR (SEQ ID NO:98), WWGGGQIVGGVYLLPRRGPRLGVRAT (SEQ ID NO:99), BRLIFLARSALIVRGSVAHKS (SEQ ID NO:100), EDLIFLARSALILRGSVAHKS (SEQ ID NO:101), BRLIFLARSALILBGRSALILRGSVAHK (SEQ ID NO:102), SAYERMCNILKGKFQTAAQRAMM (SEQ ID NO:103), SAYERNleVNILKGKFQTAAQRAVNle (SEQ ID NO:104), BRTAYERNleCNILBRGRFQTVVQBA (SEQ ID NO:105), BRIAYERMCNILLBRGKFQTAAQRA (SEQ ID NO:106), IAYERMCNILKGKFQTAAQRA (SEQ ID NO:107), LFFKClYRLFKHGLKRGPSTEGVPESM (SEQ ID NO:108), BRRLFFKTITRLFBHGLRRLLSTEGVPNSNle (SEQ ID NO:109), BRGLEPLVIAGILARRGSLVGLLHIVL (SEQ ID NO:110), BRGSDPLVVAASIVRRASIVGILHLIL (SEQ ID NO:111), RNLVPMVATVRRNLVPMVATVB (SEQ ID NO:112), RNLVPMVATVBRRNLVPMVATVB (SEQ ID NO:113), RNIVPNleVVTARRNIVPNleVVTAB (SEQ ID NO:114), PEVIPMFSALSEGATPQDLNTMLN (SEQ ID NO:115), RFIIPXFTALSGGRRALLYGATPYAIG (SEQ ID NO:116), KALGPAATLEEMMTACQGVG (SEQ ID NO:117), RRGPVVHLTLRRRGQAGDDFS (SEQ ID NO:118), RRGPVVHLTLRRRGQAGDDFS (SEQ ID NO:119), RRGPVVHLTLRGRRGQAGDDFS (SEQ ID NO:120), RRLECVYCKQQLLRREVYDFAFRDLC (SEQ ID NO:121), RRGVYDFAFRDLCRRGFAFRDLCIVYR (SEQ ID NO:122), RRGVFDYAFRDINRRGFAYRDINLAYR (SEQ ID NO:123), RRGATPVDLLGARRGALNLCLPMR (SEQ ID NO:124), RRGVTPAGLIGVRRGALQIBLPLR (SEQ ID NO:125), RGYLPAVGAPIGRRRVIRVIAHGLRLR (SEQ ID NO:196), RRSRNLGKVIDTLTCRRLMGYIPLVGA (SEQ ID NO:197), and RRIRNLGRVIETLTLNleGYIPLIGARRIRNLGRVIETLTLNleGYIPLIGAR (SEQ ID NO:199), or a fragment or variant thereof.

(235) In some embodiments the peptide according to the invention is not a peptide consisting of a sequence selected from X.sup.1-NYVTGNIPG-X.sup.3-GITFSIYLIVS; X.sup.1-IRNLGRVIETLT-X.sup.3-LNleGYIPLIGA; X.sup.1-GYLPAVGAP.sup.1-X.sup.3-VIRVIAHGLRL; X.sup.1-GGGQIIGGNYLIP-X.sup.3-PBIGVRATB; X.sup.1-NYATGNLPG-X.sup.3-GCSFSIFLLAL; X.sup.1-SRNLGKVIDTLTC-X.sup.3-LMGYIPLVGA; X.sup.1-GYIPLVGAPL-X.sup.3-VARALAHGVRV; X.sup.1-GGGQIVGGVYLLP-X.sup.3-PRLGVRATR; X.sup.1-LTFLVRSVLL.sup.1-X.sup.3-GSVLIVRGSLVH; X.sup.1-TAYERNleCNIL-X.sup.3-GRFQTVVQBA; X.sup.1-SDPLVVAASIV-X.sup.3-ASIVGILHLIL; X.sup.1-LIFLARSALIL-X.sup.3-SALILRGSVAH; X.sup.1-IAYERMCNIL-X.sup.3-GKFQTAAQRA; and X.sup.1-LEPLVIAGILA-X.sup.3-GSLVGLLHIVL; X.sup.1-NLVPMVATV-X.sup.3-NLVPMATV; X.sup.1-GYLPAVGAPIG-X.sup.3-VIRVIAHGLRL; X.sup.1-IRNLGRVIETLTG-X.sup.3-LNleGYIPLIGA; X.sup.1-GVYDFAFRDLC-X.sup.3-GFAFRDLCIVYR, X.sup.1-GVFDYAFRDIN-X.sup.3-GFAYRDINLAYR, X.sup.1-GATPVDLLGA-X.sup.3-GALNLCLPMR, X.sup.1-GVTPAGLIGV-X.sup.3-GALQIBLPLR, and X.sup.1-IRNLGRVIETLTLNleGYIPLIGA-X.sup.3-IRNLGRVIETLTLNleGYIPLIGA; optionally with an X.sup.5 in the C-terminal of the peptide, wherein X.sup.1 and X.sup.3 and X.sup.5 refers to X.sup.1, X.sup.3, and X.sup.5 of formula II.

(236) In some embodiments the peptide comprises one or more cysteine.

(237) In some embodiments the peptide contain intramolecular bonds, such as intramolecular disulfide (SS) bonds between two cys residues.

(238) In other embodiments the peptide contains intramolecular bonds, such as in the form of a acylal moiety (COOCH2-OOC. COOCHROOC or COOCR2-OOC).

(239) In some embodiments the peptide according to the present invention is not more than 58 amino acids, such as not more than 56, 54, 52, 50, 48, 46, 44, 42, 40, 38, 36, 34, 32, 30, 28, 26, 24, 22, 20, 18 amino acid residues.

(240) In some embodiments an isolated peptide according to the present invention is not a peptide consisting of a sequence of X.sup.2 or X.sup.4 as defined in table 1, table 2, or table 8.

(241) In some embodiments an isolated peptide according to the present invention comprises a sequence of X.sup.2 and/or X.sup.4 as defined in table 1, table 2, table 5, or a fragment thereof.

(242) In some embodiments the dimer peptide according to the invention consist of two identical peptide monomers.

(243) In some embodiments the immunogenic composition according to the invention is in the form of a vaccine composition.

(244) In some embodiments, the peptide of the invention comprises at most 60, at most 59, at most 58, at most 57, at most 56, at most 55, at most 54, at most 53, at most 52, at most 51, at most 50, at most 49, at most 48, at most 47, at most 46, at most 45, at most 44, at most 43, at most 42, at most 41, at most 40, at most 39, at most 38, at most 37, at most 36, at most 35, at most 34, at most 33, at most 32, at most 31, at most 30, at most 29, at most 28, at most 27, at most 26, at most 25, at most 24, at most 23, at most 22, at most 21, at most 20, at most 19, at most 18 amino acids.

(245) In some embodiments, the peptide of the invention comprises at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 31, at least 32, at least 33, at least 34, at least 35 at least 36, at least 37, at least 38, at least 39, at least 40, at least 41, at least 42, at least 43, at least 44, at least 45, at least 46, at least 47, at least 48, at least 49, at least 50, at least 51, at least 52, at least 53, at least 54, at least 55, at least 56, at least 57, at least 58, at least 59, at least 60 amino acid residues.

(246) In some embodiments, the peptide of the invention consists of 18 amino acid residues or 19 amino acid residues or 20 amino acid residues or 21 amino acid residues or 22 amino acid residues or 23 amino acid residues or 24 amino acid residues or 25 amino acid residues or 26 amino acid residues or 27 amino acid residues or 28 amino acid residues or 29 amino acid residues or 30 amino acid residues or 31 amino acid residues or 32 amino acid residues or 33 amino acid residues or 34 amino acid residues or 35 amino acid residues or 36 amino acid residuesor 37 amino acid residues or 38 amino acid residues or 39 amino acid residues or 40 amino acid residues or 41 amino acid residues or 42 amino acid residues or 43 amino acid residues or 44 amino acid residues or 45 amino acid residues or 46 amino acid residues or 47 amino acid residues or 48 amino acid residues or 49 amino acid residues or 50 amino acid residues or 51 amino acid residues or 52 amino acid residues or 53 amino acid residues or 54 amino acid residues or 55 amino acid residues or 56 amino acid residues or 57 amino acid residues or 58 amino acid residues or 59 amino acid residues or 60 amino acid residues.

(247) In some embodiments the peptide of the invention does not consist of the following sequence RFIIP[Nle]FTALSGGRRALLYGATPYAIG, where Nle denotes a nor-leucine.

(248) In some embodiments Z.sup.3, and optional Z.sup.6, Z.sup.9 and Z.sup.12 is not derived from HIV.

(249) Numbered embodiments according to the invention:

(250) 1. An isolated monomeric peptide comprising the following structure
(Z.sup.1-Z.sup.2).sub.1-Z.sup.3-(Z.sup.4-Z.sup.5).sub.2-Z.sup.6-(Z.sup.7-Z.sup.8).sub.3-Z.sup.9-(Z.sup.10-Z.sup.11).sub.4-Z.sup.12
wherein Z.sup.1, Z.sup.4, and optional Z.sup.7 and Z.sup.10 defines a linear sequence of one, two, or three arginine residues or derivatives thereof optionally followed by a glycine (G) or an alanine (A); Z.sup.2, Z.sup.5, Z.sup.8 and Z.sup.11 defines an optional amino acid selected from cysteine (C), lysine (K), aspartic acid (D), asparagine (N), glutamic acid (E), glutamine (Q), 2,3-Diaminopropionic acid (Dpr), tryptophan (W), or tyrosine (Y) or a derivative thereof; Z.sup.3, and optional Z.sup.6, Z.sup.9 and Z.sup.12 defines any chemical moiety, such as a linear amino acid sequence.
2. The isolated monomeric peptide according to embodiment 1, wherein said chemical moiety of Z.sup.3, and optional Z.sup.6, Z.sup.9 and Z.sup.12 is a linear amino acid sequence of 8-30 amino acids or a compound with our without immune modulating properties.
3. The isolated monomeric peptide according to embodiments 1 or 2, wherein Z.sup.2 defines an amino acid selected from cysteine (C), lysine (K), aspartic acid (D), asparagine (N), glutamic acid (E), glutamine (Q), 2,3-Diaminopropionic acid (Dpr), tryptophan (W), or tyrosine (Y) or a derivative thereof.
4. The isolated monomeric peptide according to any one of embodiments 1-3, wherein Z.sup.5 defines an amino acid selected from cysteine (C), lysine (K), aspartic acid (D), asparagine (N), glutamic acid (E), glutamine (Q), 2,3-Diaminopropionic acid (Dpr), tryptophan (W), or tyrosine (Y) or a derivative thereof.
5. The isolated monomeric peptide according to embodiments 1-4, wherein Z.sup.8 defines an amino acid selected from cysteine (C), lysine (K), aspartic acid (D), asparagine (N), glutamic acid (E), glutamine (Q), 2,3-Diaminopropionic acid (Dpr), tryptophan (W), or tyrosine (Y) or a derivative thereof.
6. The isolated monomeric peptide according to embodiments 1-5, wherein Z.sup.11 defines an amino acid selected from cysteine (C), lysine (K), aspartic acid (D), asparagine (N), glutamic acid (E), glutamine (Q), 2,3-Diaminopropionic acid (Dpr), tryptophan (W), or tyrosine (Y) or a derivative thereof.
7. The isolated monomeric peptide according to any one of embodiments 1-6, wherein Z.sup.7 defines a linear sequence of one, two, or three arginine residues or derivatives thereof optionally followed by a glycine (G) or an alanine (A).
8. The isolated monomeric peptide according to any one of embodiments 1-7, wherein Z.sup.10 defines a linear sequence of one, two, or three arginine residues or derivatives thereof optionally followed by a glycine (G) or an alanine (A).
9. The isolated monomeric peptide according to any one of embodiments 1-8, wherein Z.sup.6 defines any chemical moiety, such as a linear amino acid sequence.
10. The isolated monomeric peptide according to any one of embodiments 1-9, wherein Z.sup.9 defines any chemical moiety, such as a linear amino acid sequence.
11. The isolated monomeric peptide according to any one of embodiments 1-10, wherein Z.sup.12 defines any chemical moiety, such as a linear amino acid sequence.
12. The isolated monomeric peptide according to any one of embodiments 1-11, wherein Z.sup.1, Z.sup.4, and optional Z.sup.7 and Z.sup.10 is followed by a glycine (G) or an alanine (A).
13. The isolated monomeric peptide according to any one of embodiments 1-12, wherein Z.sup.3, and optional Z.sup.6, Z.sup.9 and Z.sup.12 is a linear amino acid sequence of 8-30 amino acids derived from an antigen with more than 40%, such as more than 45%, such as more than 50%, such as more than 55%, such as more than 60%, such as more than 65%, such as more than 70%, such as more than 75%, such as more than 80%, such as more than 85%, such as more than 90%, such as more than 95%, such as more than 96%, such as more than 97%, such as more than 98%, such as more than 99%, such as 100% sequence identity to a specific natural antigen.
14. The isolated monomeric peptide according to any one of embodiments 1-13, wherein Z.sup.3, and optional Z.sup.6, Z.sup.9 and Z.sup.12 defines a specific natural antigen of a protein or peptide sequence derived from a disease antigen, such as an infectious agent, such as bacteria, virus, parasite, fungus, or cancer antigens such as oncogene (lung, stomach, breast cancer) or an antigen causing an autoimmune disease such as diabetes, multiple sclerosis (MS), celiac disease, Myalgic Encephalomyelitis (ME), psoriasis, and/or Crohn's Disease.
15. The isolated monomeric peptide according to embodiment 14, wherein said specific natural antigen is a viral protein, such as a structural protein, such as a capsid protein, a regulatory protein, an enzymatic protein, and a proteolytic protein.
16. The isolated monomeric peptide according to any one of embodiments 14-15, wherein said viral protein is selected from a core protein or an envelope protein, of a virus selected from the Hepatitis C virus, influenza virus, such as an M2 protein, human immunodeficiency virus (HIV), cytomegalovirus (CMV), and Human papillomavirus (HPV).
17. The isolated monomeric peptide according to embodiment 16, wherein said viral protein is a viral protein of Hepatitis C virus selected from any one HCV consensus sequence of a specific genotype, such as 1, such as subtypes 1a and 1b, genotype 2, such as 2a and 2b, genotype 3, such as 3a, genotype 4, genotype 5, and genotype 6.
18. The isolated monomeric peptide according to any one of embodiments 1-17, wherein a sequence of amino acids defined by (Z.sup.1-Z.sup.2).sub.1-Z.sup.3-(Z.sup.4-Z.sup.5).sub.2-Z.sup.6-(Z.sup.7-Z.sup.8).sub.3-Z.sup.9-(Z.sup.10-Z.sup.11).sub.4-Z.sup.12 is not found in the native sequence of a natural antigen.
19. The isolated monomeric peptide according to any one of embodiments 1-18, which monomeric peptide is of 10-60 amino acids, such as of 11-60 amino acids, such as of 12-60 amino acids, such as of 13-60 amino acids, such as of 14-60 amino acids, such as of 15-60 amino acids, such as of 16-60 amino acids, such as of 17-60 amino acids, such as of 18-60 amino acids, such as of 19-60 amino acids, such as of 20-60 amino acids, such as of 21-60 amino acids, such as of 22-60 amino acids, such as of 23-60 amino acids, such as of 24-60 amino acids, such as of 25-60 amino acids, such as of 26-60 amino acids, such as of 27-60 amino acids, such as of 28-60 amino acids, such as of 29-60 amino acids, such as of 30-60 amino acids, such as of 31-60 amino acids, such as of 32-60 amino acids, such as of 33-60 amino acids, such as of 34-60 amino acids, such as of 35-60 amino acids, such as of 36-60 amino acids, such as of 37-60 amino acids, such as of 38-60 amino acids, such as of 39-60 amino acids, such as of 40-60 amino acids, such as of 42-60 amino acids, such as of 44-60 amino acids, such as of 46-60 amino acids, such as of 48-60 amino acids, such as of 50-60 amino acids, such as of 52-60 amino acids, such as of 54-60 amino acids, such as of 56-60 amino acids, such as of 58-60 amino acids.
20. The isolated monomeric peptide according to any one of embodiments 1-19, which monomeric peptide is of 10-60 amino acids, such as 10-58 amino acids, such as 10-56 amino acids, such as 10-54 amino acids, such as 10-52 amino acids, such as 10-50 amino acids, such as 10-48 amino acids, such as 10-46 amino acids, such as 10-44 amino acids, such as 10-42 amino acids, such as 10-40 amino acids, such as 10-39 amino acids, such as 10-38 amino acids, such as 10-37 amino acids, such as 10-36 amino acids, such as 10-35 amino acids, such as 10-34 amino acids, such as 10-33 amino acids, such as 10-32 amino acids, such as 10-31 amino acids, such as 10-30 amino acids, such as 10-29 amino acids, such as 10-28 amino acids, such as 10-27 amino acids, such as 10-26 amino acids, such as 10-25 amino acids, such as 10-24 amino acids, such as 10-23 amino acids, such as 10-22 amino acids, such as 10-21 amino acids, such as 10-20 amino acids, such as 10-19 amino acids, such as 10-18 amino acids, such as 10-17 amino acids, such as 10-16 amino acids, such as 10-15 amino acids, such as 10-14 amino acids, such as 10-13 amino acids, such as 10-12 amino acids, such as 10-11 amino acids.
21. The isolated monomeric peptide according to any one of embodiments 1-20, which monomeric peptide consist of not more than about 55 amino acids, such as not more than about 50 amino acids, such as not more than about 45 amino acids, such as not more than about 40 amino acids, such as not more than about 38 amino acids, such as not more than about 36 amino acids, such as not more than about 34 amino acids, such as not more than about 32 amino acids, such as not more than about 30 amino acids, such as not more than about 28 amino acids, such as not more than about 26 amino acids, such as not more than about 24 amino acids, such as not more than about 22 amino acids, such as not more than about 20 amino acids, such as not more than about 18 amino acids, such as not more than about 16 amino acids, such as not more than about 14 amino acids, such as not more than about 12 amino acids, such as not more than about 10 amino acids.
22. The isolated monomeric peptide according to any one of embodiments 1-21, which monomeric peptide consist of at least about 10 amino acids, such as at least about 12 amino acids, such as at least about 14 amino acids, such as at least about 16 amino acids, such as at least about 18 amino acids, such as at least about 20 amino acids, such as at least about 22 amino acids, such as at least about 24 amino acids, such as at least about 26 amino acids, such as at least about 28 amino acids, such as at least about 30 amino acids, such as at least about 32 amino acids, such as at least about 34 amino acids, such as at least about 36 amino acids, such as at least about 38 amino acids, such as at least about 40 amino acids, such as at least about 45 amino acids, such as at least about 50 amino acids, such as at least about 55 amino acids, such as at least about 60.
23. The isolated monomeric peptide according to any one of embodiments 1-22, wherein the overall net charge of (Z.sup.1-Z.sup.2).sub.1-Z.sup.3-(Z.sup.4-Z.sup.5).sub.2-Z.sup.6-(Z.sup.7-Z.sup.8).sub.3-Z.sup.9-(Z.sup.10-Z.sup.11).sub.4-Z.sup.12 is equal to or above 0, such as above 1, 2, 3, 4, or 5.
24. The isolated monomeric peptide according to any one of embodiments 1-23, wherein said monomeric peptide is capable of inducing a humoral immune response.
25. The isolated monomeric peptide according to any one of embodiments 1-24, wherein said monomeric peptide comprises at least one amino acid selected from a Cys, a Lys, an Asp, and a Glu residue, or derivatives thereof.
26. The isolated monomeric peptide according to any one of embodiments 1-25, which monomeric peptide contain one or more intramolecular bond, such as one or more Cys-Cys bond.
27. The isolated monomeric peptide according to any one of embodiments 1-26, which monomeric peptide has delayed proteolytic degradation in the N-terminal, such as by incorporation of the first 1, 2, or 3 amino acids in the N-terminal in the D-form, or by incorporation of the first 1, 2, or 3 amino acids in the N-terminal in beta or gamma form.
28. The isolated peptide according to any one of embodiment 1-27, wherein said peptide is demonstrated to translocate across a plasma membrane in the assay based on biotinylation of peptides as described in example 5.
29. The isolated peptide according to any one of embodiments 1-28, wherein said peptide is capable of inducing a T lymphocyte response.
30. The isolated peptide according to any one of embodiments 1-29, wherein the net charge of Z.sup.3, and/or optional Z.sup.6, Z.sup.9 and Z.sup.12 is below or equal to 0.
31. The isolated peptide according to any one of embodiments 1-30, wherein the net charge of Z.sup.3 is below or equal to 0; and wherein the net charge of Z.sup.6 and/or optional Z.sup.9 and Z.sup.12 is above or equal to 1.
32. The isolated peptide according to any one of embodiments 1-31, wherein the net charge of Z.sup.3, and/or optional Z.sup.6, Z.sup.9 and Z.sup.12 are above or equal to 1.
33. The isolated peptide according to any one of embodiments 1-32, wherein the net charge of Z.sup.3 is above or equal to 1; and wherein the net charge of Z.sup.6 and/or optional Z.sup.9 and Z.sup.12 is below or equal to 0.
34. The isolated peptide according to any one of embodiments 1-33, wherein the peptide comprises one or more cysteine.
35. The isolated peptide according to any one of embodiments 1-34, wherein the N- and/or C-terminal amino acid in Z.sup.3, and/or optional Z.sup.6, Z.sup.9 and Z.sup.12 is a hydrophilic or polar amino acid.
36. The isolated peptide according to any one of embodiments 1-35, wherein Z.sup.3, and/or optional Z.sup.6, Z.sup.9 and Z.sup.12 defines a sequence of 8-25 amino acids, such as 8-20 amino acids, such as 8-15 amino acids.
37. The isolated peptide according to any one of embodiments 1-36, wherein Z.sup.3, and/or optional Z.sup.6, Z.sup.9 and Z.sup.12 defines a sequence of less than 25, such as less than 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7 or 6 amino acids.
38. The isolated peptide according to any one of embodiments 1-37, wherein Z.sup.3, and/or optional Z.sup.6, Z.sup.9 and Z.sup.12 defines a sequence of more than 8, such as more than 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 amino acids.
39. The isolated peptide according to any one of embodiments 1-38, which does not consist of the following sequence RFIIP[Nle]FTALSGGRRALLYGATPYAIG, where Nle denotes a nor-leucine.
40. The isolated peptide according to any one of embodiments 1-39, wherein Z.sup.3, and/or optional Z.sup.6, Z.sup.9 and Z.sup.12 is not derived from HIV.
41. The isolated peptide according to any one of embodiments 1-40, wherein Z.sup.3, and/or optional Z.sup.6, Z.sup.9 and Z.sup.12 is a linear sequence of less than 12 amino acids.
42. The isolated peptide according to any one of embodiments 1-41, wherein Z.sup.3, and/or optional Z.sup.6, Z.sup.9 and Z.sup.12 is a linear sequence of less than 12 amino acids.
43. The isolated peptide according to any one of embodiments 1-42, wherein Z.sup.3, and/or optional Z.sup.6, Z.sup.9 and Z.sup.12 do not contain nor-leucine.
44. The isolated peptide according to any one of embodiments 1-43, wherein Z.sup.3, and/or optional Z.sup.6, Z.sup.9 and Z.sup.12 do not contain nor-leucine.
45. The isolated peptide according to any one of embodiments 1-44, wherein Z.sup.3, and/or optional Z.sup.6, Z.sup.9 and Z.sup.12 only contains natural amino acids.
46. The isolated peptide according to any one of embodiments 1-45, wherein Z.sup.3, and/or optional Z.sup.6, Z.sup.9 and Z.sup.12 only contains natural amino acids.
47. The isolated peptide according to any one of embodiments 1-46, wherein Z.sup.3, and/or optional Z.sup.6, Z.sup.9 and Z.sup.12 only contains natural amino acids if derived from HIV.
48. The isolated peptide according to any one of embodiments 1-47, wherein Z.sup.3, and/or optional Z.sup.6, Z.sup.9 and Z.sup.12 is derived from HCV, CMV, HPV, Influenza, adenoviruses, herpesviruses, or picornaviruses.
49. The isolated peptide according to any one of embodiments 1-48, wherein Z.sup.1 is as defined in any one of table 3, table 4, table 5, or table 7, such as any one selected from R, RR, RRR, RG, RRG and RRRG.
50. The isolated peptide according to any one of embodiments 1-49, wherein Z.sup.2 is as defined in any one of table 3, table 4, table 5, or table 7, such as any one selected from Dpr(Aoa), C, K, Lys(Me), D, E, Dpr(Ser).
51. The isolated peptide according to any one of embodiments 1-50, wherein Z.sup.3 is as defined in any one of table 3, table 4, table 5, or table 7, such as any one selected from GGQLIGGIYLIPG (SEQ ID NO:313), VITYSIFLIVS (SEQ ID NO:314), TANWARVIS (SEQ ID NO:315), GYLPAVGAPI (SEQ ID NO:316), NIVPZVVTA (SEQ ID NO:317), VTPADLIGA (SEQ ID NO:318), PRPEGYTLFF (SEQ ID NO:319), LPYPRGYTLFV (SEQ ID NO:320), ETILTPRDV (SEQ ID NO:321), SSTSPVYDL (SEQ ID NO:322), TAYERZCNIL (SEQ ID NO:323), TVIGASZIPLL (SEQ ID NO:324), AAFEEZXITS (SEQ ID NO:325), GLEPLVIAGILA (SEQ ID NO:326), TAFLVRNVA (SEQ ID NO:327), TPI(Har)QDWGNRAN (SEQ ID NO:328), TPT(Har)NGWDVKLS (SEQ ID NO:329), LECVYCKQQLL (SEQ ID NO:330), GVYDFAFRDLC (SEQ ID NO:331), GVFDYAFRDIN (SEQ ID NO:332), and VDIRTLEDLL (SEQ ID NO:333).
52. The isolated peptide according to any one of embodiments 1-51, wherein Z.sup.4 is as defined in any one of table 3, table 4, table 5, or table 7, such as any one selected from R, RR, RRR, RG, RRG and RRRG.
53. The isolated peptide according to any one of embodiments 1-52, wherein Z.sup.5 is as defined in any one of table 3, table 4, table 5, or table 7, such as any one selected from Dpr(Aoa), C, K, Lys(Me), D, E, Dpr(Ser).
54. The isolated peptide according to any one of embodiments 1-53, wherein Z.sup.6 is as defined in any one of table 3, table 4, table 5, or table 7, such as any one selected from EVYDFAFRDLC (SEQ ID NO:334), GFAFRDLCIVY (SEQ ID NO:335), GFAYRDINLAY (SEQ ID NO:336), GTLGIVCPIG (SEQ ID NO:337), GLEPLVIAGILA (SEQ ID NO:338), TPIXQDWENRAN (SEQ ID NO:339), VAFEDLXZZSFI (SEQ ID NO:340), RFQTVVQBA (SEQ ID NO:341), GSLVGLLHIVL (SEQ ID NO:342), SIARSVTIZXASVVH (SEQ ID NO:343), TPTRQEWDCRIS (SEQ ID NO:344), TPTRQEWDARIS (SEQ ID NO:345), TPI(Har)QEW(Har)SL(Nle)NQEW (SEQ ID NO:346), IGDLIVAQV (SEQ ID NO:347), QYNPVAVZF (SEQ ID NO:348), GYTLFFTS (SEQ ID NO:349), GYTLFVSD (SEQ ID NO:350), NTLZTPRDV (SEQ ID NO:351), SSTSPVYNL (SEQ ID NO:352), VITFSIYLIVS (SEQ ID NO:353), GGNVIGGIYZIPR (SEQ ID NO:354), ANWAKVIL (SEQ ID NO:355), VIRVIAHGLRL (SEQ ID NO:356), and IGDLIVQAV (SEQ ID NO:478).
55. The isolated peptide according to any one of embodiments 1-54, wherein Z.sup.7 is as defined in any one of table 3, table 4, table 5, or table 7, such as any one selected from R, RR, RRR, RG, RRG and RRRG.
56. The isolated peptide according to any one of embodiments 1-55, wherein Z.sup.8 is as defined in any one of table 3, table 4, table 5, or table 7, such as any one selected from Dpr(Aoa), C, K, Lys(Me), D, E, Dpr(Ser).
57. The isolated peptide according to any one of embodiments 1-56, wherein Z.sup.9 is as defined in any one of table 3, table 4, table 5, or table 7, such as NWAKVI.
58. The isolated peptide according to any one of embodiments 1-57, which peptide consist of (Z.sup.1-Z.sup.2).sub.1-Z.sup.3-(Z.sup.4-Z.sup.5).sub.2-Z.sup.6-(Z.sup.7-Z.sup.8).sub.3-Z.sup.9 as defined in any one of table 3, table 4, table 5, or table 7, such as any one selected from RRGGQLIGGIYLIPGRRVITFSIYLIVS (SEQ ID NO:357), RRRGGQLIGGIYLIPGRRVITFSIYLIVS (SEQ ID NO:358), RRGGQLIGGIYLIPGRRRVITFSIYLIVS (SEQ ID NO:359), RRGGQLIGGIYLIPGRRVITFSIYLIVSR (SEQ ID NO:360), RRGGQLIGGIYLIPGRRVITFSIYLIVSRR (SEQ ID NO:361), RRVITYSIFLIVSRRGGNVIGGIYZIPR (SEQ ID NO:362), RRRVITYSIFLIVSRRGGNVIGGIYZIPR (SEQ ID NO:363), RRVITYSIFLIVSRRRGGNVIGGIYZIPR (SEQ ID NO:364), RRRVITYSIFLIVSRRRGGNVIGGIYZIPR (SEQ ID NO:365), RRGTANWARVISRANWAKVILRNWAKVI (SEQ ID NO:366), RGTANWARVISRRANWAKVILRNWAKVI (SEQ ID NO:367), RGTANWARVISRANWAKVILRNWAKVI (SEQ ID NO:368), RGTANWARVISRGANWAKVILRNWAKVI (SEQ ID NO:369), RRGTANWARVISRANWARVILRNWAKVI (SEQ ID NO:370), RGTANWARVISRRANWARVILRNWAKVI (SEQ ID NO:371), RGTANWARVISRANWARVILRNWAKVI (SEQ ID NO:372), RGTANWARVISRGANWARVILRNWAKVI (SEQ ID NO:373), RGYLPAVGAPIRRRVIRVIAHGLRLR (SEQ ID NO:374), RRGYLPAVGAPIRRVIRVIAHGLRLR (SEQ ID NO:375), RRGYLPAVGAPIRRRVIRVIAHGLRL (SEQ ID NO:376), RRGYLPAVGAPIRRVIRVIAHGLRL (SEQ ID NO:377), RGYLPAVGAPIRRVIRVIAHGLRLR (SEQ ID NO:378), RGYLPAVGAPIRVIRVIAHGLRLR (SEQ ID NO:379), RGYLPAVGAPIRRVIRVIAHGLRL (SEQ ID NO:380), RGNIVPZVVTARRIGDLIVAQV (SEQ ID NO:381), RRNIVPZVVTARRIGDLIVAQV (SEQ ID NO:382), RRRNIVPZVVTARRIGDLIVAQV (SEQ ID NO:383), RRNIVPZVVTARRRIGDLIVAQV (SEQ ID NO:384), RGVTPADLIGARRQYNPVAVZF (SEQ ID NO:385), RRVTPADLIGARRQYNPVAVZF (SEQ ID NO:386), RRRVTPADLIGARRQYNPVAVZF (SEQ ID NO:387), RRVTPADLIGARRRQYNPVAVZF (SEQ ID NO:388), RRGPRPEGYTLFFRGYTLFFTSR (SEQ ID NO:389), RGPRPEGYTLFFRRGYTLFFTSR (SEQ ID NO:390), RRGPRPEGYTLFFRRGYTLFFTSR (SEQ ID NO:391), RRGPRPEGYTLFFRRRGYTLFFTSR (SEQ ID NO:392), RRRGPRPEGYTLFFRRGYTLFFTSR (SEQ ID NO:393), RGLPYPRGYTLFVRRGYTLFVSDR (SEQ ID NO:394), RRGLPYPRGYTLFVRRGYTLFVSDR (SEQ ID NO:395), RRGLPYPRGYTLFVRRRGYTLFVSDR (SEQ ID NO:396), RRRGLPYPRGYTLFVRRGYTLFVSDR (SEQ ID NO:397), RRGLPYPRGYTLFVRRGYTLFVSDR (SEQ ID NO:398), RRGETILTPRDVRNTLZTPRDVR (SEQ ID NO:399), RGETILTPRDVRRNTLZTPRDVR (SEQ ID NO:400), RGETILTPRDVRNTLZTPRDVR (SEQ ID NO:401), RGETILTPRDVRGNTLZTPRDVR (SEQ ID NO:402), RRSSTSPVYDLRRSSTSPVYNLR (SEQ ID NO:403), RRSSTSPVYDLRRRSSTSPVYNLR (SEQ ID NO:404), RRRSSTSPVYDLRRSSTSPVYNLR (SEQ ID NO:405), RRRSSTSPVYDLRRRSSTSPVYNLR (SEQ ID NO:406), RRTAYERZCNILRRGLEPLVIAGILA (SEQ ID NO:407), RRRTAYERZCNILRRGLEPLVIAGILA (SEQ ID NO:408), RRTAYERZCNILRRRGLEPLVIAGILA (SEQ ID NO:409), RRTAYERZCNILRRGLEPLVIAGILAR (SEQ ID NO:410), RRTAYERZCNILRRGLEPLVIAGILARR (SEQ ID NO:411), RRTVIGASZIPLLRGTPIXQDWENRAN (SEQ ID NO:412), RRRTVIGASZIPLLRGTPIXQDWENRAN (SEQ ID NO:413), RRTVIGASZIPLLRRGTPIXQDWENRAN (SEQ ID NO:414), RRRTVIGASZIPLLRRGTPIXQDWENRAN (SEQ ID NO:415), RRRTVIGASZIPLLRRGTPIXQDWENRANR (SEQ ID NO:416), RRAAFEEZXITSRRVAFEDLXZZSFI (SEQ ID NO:417), RRRAAFEEZXITSRRVAFEDLXZZSFI (SEQ ID NO:418), RRRAAFEEZXITSRRGVAFEDLXZZSFI (SEQ ID NO:419), RRRAAFEEZXITSRRRVAFEDLXZZSFI (SEQ ID NO:420), RRRAAFEEZXITSRRRVAFEDLXZZSFIGR (SEQ ID NO:421), RRTAYERZCNILRRGRFQTVVQBA (SEQ ID NO:422), RRTAYERZCNILRRGRFQTVVQBAR (SEQ ID NO:423), RTAYERZCNILRGRFQTVVQBAR (SEQ ID NO:424), RRTAYERZCNILRGRFQTVVQBA (SEQ ID NO:425), BRGLEPLVIAGILARRGSLVGLLHIVL (SEQ ID NO:426), RRGLEPLVIAGILARRGSLVGLLHIVL (SEQ ID NO:427), RRGLEPLVIAGILARRGSLVGLLHIVLR (SEQ ID NO:428), RRGLEPLVIAGILARRRGSLVGLLHIVL (SEQ ID NO:429), RRGLEPLVIAGILARRRGSLVGLLHIVLR (SEQ ID NO:430), RTAFLVRNVARSIARSVTIZXASVVH (SEQ ID NO:431), RTAFLVRNVARRSIARSVTIZXASVVH (SEQ ID NO:432), RRTAFLVRNVARSIARSVTIZXASVVH (SEQ ID NO:433), RRTAFLVRNVARRSIARSVTIZXASVVH (SEQ ID NO:434), RRTAFLVRNVARRSIARSVTIZXASVVHR (SEQ ID NO:435), RRTAFLVRNVARRSIARSVTIZXASVVHRR (SEQ ID NO:436), RGDpr(Aoa)TPI(Har)QDWGNRANRGTPTRQEWDCRIS (SEQ ID NO:437), RGDpr(Aoa)TPI(Har)QDWGNRANRGTPTRQEWDARIS (SEQ ID NO:438), RGTPI(Har)QDWGNRANRGTPTRQEWDCRIS (SEQ ID NO:439), RGTPI(Har)QDWGNRANRGTPTRQEWDARIS (SEQ ID NO:440), RGCTPI(Har)QDWGNRANRGTPTRQEWDCRIS (SEQ ID NO:441), RGCTPI(Har)QDWGNRANRGTPTRQEWDARIS (SEQ ID NO:442), RGKTPI(Har)QDWGNRANRGTPTRQEWDCRIS (SEQ ID NO:443), RGKTPI(Har)QDWGNRANRGTPTRQEWDARIS (SEQ ID NO:444), RGLys(Me)TPI(Har)QDWGNRANRGTPTRQEWDCRIS (SEQ ID NO:445), RGLys(Me)TPI(Har)QDWGNRANRGTPTRQEWDARIS (SEQ ID NO:446), RGDTPI(Har)QDWGNRANRGTPTRQEWDCRIS (SEQ ID NO:447), RGDTPI(Har)QDWGNRANRGTPTRQEWDARIS (SEQ ID NO:448), RGETPI(Har)QDWGNRANRGTPTRQEWDCRIS (SEQ ID NO:449), RGETPI(Har)QDWGNRANRGTPTRQEWDARIS (SEQ ID NO:450), RGDpr(Ser)TPT(Har)NGWDVKLSRGTPI(Har)QEW(Har)SL(Nle)NQEW (SEQ ID NO:451), RGTPT(Har)NGWDVKLSRGTPI(Har)QEW(Har)SL(Nle)NQEW (SEQ ID NO:452), RGKTPT(Har)NGWDVKLSRGTPI(Har)QEW(Har)SL(Nle)NQEW (SEQ ID NO:453), RGCTPT(Har)NGWDVKLSRGTPI(Har)QEW(Har)SL(Nle)NQEW (SEQ ID NO:454), RGLys(Me)TPT(Har)NGWDVKLSRGTPI(Har)QEW(Har)SL(Nle)NQEW (SEQ ID NO:455), RGDTPT(Har)NGWDVKLSRGTPI(Har)QEW(Har)SL(Nle)NQEW (SEQ ID NO:456), RGETPT(Har)NGWDVKLSRGTPI(Har)QEW(Har)SL(Nle)NQEW (SEQ ID NO:457), RRLECVYCKQQLLRREVYDFAFRDLC (SEQ ID NO:458), RRLECVYCKQQLLRRRGEVYDFAFRDLC (SEQ ID NO:459), RRRLECVYCKQQLLRRGEVYDFAFRDLC (SEQ ID NO:460), RRRLECVYCKQQLLRRRGEVYDFAFRDLC (SEQ ID NO:461), RRRGLECVYCKQQLLRRRGEVYDFAFRDLC (SEQ ID NO:462), RRGVYDFAFRDLCRRGFAFRDLCIVYR (SEQ ID NO:463), RRGVYDFAFRDLCRRRGGFAFRDLCIVY (SEQ ID NO:464), RRRGVYDFAFRDLCRRGGFAFRDLCIVYR (SEQ ID NO:465), RRRGVYDFAFRDLCRRRGGFAFRDLCIVY (SEQ ID NO:466), RRRGGVYDFAFRDLCRRRGGFAFRDLCIVYR (SEQ ID NO:467), RRGVFDYAFRDINRRGFAYRDINLAYR (SEQ ID NO:468), RRGVYDFAFRDLCRRRGGFAFRDLCIVY (SEQ ID NO:469), RRRGVYDFAFRDLCRRGGFAFRDLCIVYR (SEQ ID NO:470), RRRGVYDFAFRDLCRRRGGFAFRDLCIVY (SEQ ID NO:471), RRRGGVYDFAFRDLCRRRGGFAFRDLCIVYR (SEQ ID NO:472), RRVDIRTLEDLLRRGTLGIVCPIGR (SEQ ID NO:473), RRVDIRTLEDLLRRRGGTLGIVCPIG (SEQ ID NO:474), RRRVDIRTLEDLLRRGGTLGIVCPIGR (SEQ ID NO:475), RRRVDIRTLEDLLRRRGGTLGIVCPIG (SEQ ID NO:476), RRRGVDIRTLEDLLRRRGGTLGIVCPIGR (SEQ ID NO:477), RGNIVPZVVTARRIGDLIVQAV (SEQ ID NO:479), RRNIVPZVVTARRIGDLIVQAV (SEQ ID NO:480), RRRNIVPZVVTARRIGDLIVQAV (SEQ ID NO:481), and RRNIVPZVVTARRRIGDLIVQAV (SEQ ID NO:482).
59. The isolated peptide according to any one of embodiments 1-58, which peptide is not specifically disclosed in any one PCT application with application numbers WO2000N000075, WO2011DK050460, or WO2012DK050010.
60. The isolated peptide according to any one of embodiments 1-59, which peptide is not a peptide selected from RRGYIPLVGAPLGBGRVARALAHGVRV (SEQ ID NO:47), RGYIPLVGAPLGRRVARALAHGVRV (SEQ ID NO:48), RGYIPLVGAPLGRRRVARALAHGVRVR (SEQ ID NO:49), RRGYIPLVGAPLGRRVARALAHGVRV (SEQ ID NO:50), RRGYIPLVGAPLGRRRVARALAHGVRV (SEQ ID NO:51), BRGYIPLVGAPLGRRVARALAHGVRV (SEQ ID NO:52), RRRGYIPLVGAPLGBRVARALAHGVRV (SEQ ID NO:53), RGYIPLVGAPLGKKKVARALAHGVRV (SEQ ID NO:54), RGYIPLVGAPLGRRRVARALAHGVRV (SEQ ID NO:55), KKGYIPLVGAPLGKKVARALAHGVRV (SEQ ID NO:56), WGYIPLVGAPLGRRVARALAHGVRV (SEQ ID NO:57), WWGYIPLVGAPLGRRVARALAHGVRV (SEQ ID NO:58), EEGYIPLVGAPLGEEVARALAHGVRV (SEQ ID NO:59), GGGYIPLVGAPLGGGVARALAHGVRV (SEQ ID NO:60), EEGYIPLVGAPLGRRVARALAHGVRV (SEQ ID NO:61), RRGYIPLVGAPLGLRRVARALAHGVRV (SEQ ID NO:62), WWGYIPLVGAPLGRRVARALAHGVRV (SEQ ID NO:63), WWGYIPLVGAPLGRRRVARALAHGVRV (SEQ ID NO:64), WWGYIPLVGAPLGRVARALAHGVRV (SEQ ID NO:65), RGYIPLVGAPLGRRVARALAHGVRV (SEQ ID NO:66), RRGYLPAVGAPIGBRVIRVIAHGLRL (SEQ ID NO:67), RRGYIPLVGAPLGBRVARALAHGVRV (SEQ ID NO:68), GYIPLVGAPLGGVARALAHGVRV (SEQ ID NO:69), WWGYLPAVGAPIRRVIRVIAHGLRL (SEQ ID NO:70), GYIPLVGAPLGGVARALAHGVRV (SEQ ID NO:71), RRGYIPLVGAPLGBGRVARALAHGVRV (SEQ ID NO:72), RGYIPLVGAPLGRRVARALAHGVRV (SEQ ID NO:73), RGYIPLVGAPLGRRRVARALAHGVRV (SEQ ID NO:74), RRGYIPLVGAPLGRRVARALAHGVRV (SEQ ID NO:75), RRGYIPLVGAPLGRRRVARALAHGVRV (SEQ ID NO:76), BRGYIPLVGAPLGRRVARALAHGVRV (SEQ ID NO:77), RRRGYIPLVGAPLGBRVARALAHGVRV (SEQ ID NO:78), RGYIPLVGAPLGKKKVARALAHGVRV (SEQ ID NO:79), RGYIPLVGAPLGRRRVARALAHGVRV (SEQ ID NO:80), KKGYIPLVGAPLGKKVARALAHGVRV (SEQ ID NO:81), WGYIPLVGAPLGRRVARALAHGVRV (SEQ ID NO:82), WWGYIPLVGAPLGRRVARALAHGVRV (SEQ ID NO:83), RRGYIPLVGAPLGLRRVARALAHGVRV (SEQ ID NO:84), RRNYVTGNIPGBRGITFSIFLIVS (SEQ ID NO:85), WWNYATGNLPGRRCSFSIFLLAL (SEQ ID NO:86), WWNYVTGNIPGBRGITFSIFLIVS (SEQ ID NO:87), WWNYVTGNIPGRRGITFSIFLIVS (SEQ ID NO:88), RRNYATGNLPGRRGCSFSIFLLAL (SEQ ID NO:89), RRVTGNIPGSTYSGBRGITFSIYLIVS (SEQ ID NO:90), RRIRNLGRVIETLTGBRLNleGYIPLIGA (SEQ ID NO:91), RRSRNLGKVIDTLTCBRLMGYIPLVGA (SEQ ID NO:92), SRNLGKVIDTLTCGFADLMGYIPLVGA (SEQ ID NO:93), WWIRNLGRVIETLTRRLNleGYIPLIGA (SEQ ID NO:94), WWSRNLGKVIDTLTCRRLMGYIPLVGA (SEQ ID NO:95), RRGGGQIIGGNYLIPRBPBIGVRATB (SEQ ID NO:96), GGGQIVGGVYLLPRRGPRLGVRATR (SEQ ID NO:97), RRGGGQIVGGVYLLPRRGPRLGVRATR (SEQ ID NO:98), WWGGGQIVGGVYLLPRRGPRLGVRAT (SEQ ID NO:99), BRLIFLARSALIVRGSVAHKS (SEQ ID NO:100), EDLIFLARSALILRGSVAHKS (SEQ ID NO:101), BRLIFLARSALILBGRSALILRGSVAHK (SEQ ID NO:102), SAYERMCNILKGKFQTAAQRAMM (SEQ ID NO:103), SAYERNleVNILKGKFQTAAQRAVNle (SEQ ID NO:104), BRTAYERNleCNILBRGRFQTVVQBA (SEQ ID NO:105), BRIAYERMCNILLBRGKFQTAAQRA (SEQ ID NO:106), IAYERMCNILKGKFQTAAQRA (SEQ ID NO:107), LFFKClYRLFKHGLKRGPSTEGVPESM (SEQ ID NO:108), BRRLFFKTITRLFBHGLRRLLSTEGVPNSNle (SEQ ID NO:109), BRGLEPLVIAGILARRGSLVGLLHIVL (SEQ ID NO:110), BRGSDPLVVAASIVRRASIVGILHLIL (SEQ ID NO:111), RNLVPMVATVRRNLVPMVATVB (SEQ ID NO:112), RNLVPMVATVBRRNLVPMVATVB (SEQ ID NO:113), RNIVPNleVVTARRNIVPNleVVTAB (SEQ ID NO:114), PEVIPMFSALSEGATPQDLNTMLN (SEQ ID NO:115), RFIIPXFTALSGGRRALLYGATPYAIG (SEQ ID NO:116), KALGPAATLEEMMTACQGVG (SEQ ID NO:117), RRGPVVHLTLRRRGQAGDDFS (SEQ ID NO:118), RRGPVVHLTLRRRGQAGDDFS (SEQ ID NO:119), RRGPVVHLTLRGRRGQAGDDFS (SEQ ID NO:120), RRLECVYCKQQLLRREVYDFAFRDLC (SEQ ID NO:121), RRGVYDFAFRDLCRRGFAFRDLCIVYR (SEQ ID NO:122), RRGVFDYAFRDINRRGFAYRDINLAYR (SEQ ID NO:123), RRGATPVDLLGARRGALNLCLPMR (SEQ ID NO:124), RRGVTPAGLIGVRRGALQIBLPLR (SEQ ID NO:125), RGYLPAVGAPIGRRRVIRVIAHGLRLR (SEQ ID NO:196), RRSRNLGKVIDTLTCRRLMGYIPLVGA (SEQ ID NO:197), RRIRNLGRVIETLTLNleGYIPLIGARRIRNLGRVIETLTLNleGYIPLIGAR (SEQ ID NO:199), X.sup.1-NYVTGNIPG-X.sup.3-GITFSIYLIVS; X.sup.1-IRNLGRVIETLT-X.sup.3-LNleGYIPLIGA; X.sup.1-GYLPAVGAPI-X.sup.3-VIRVIAHGLRL; X.sup.1-GGGQIIGGNYLIP-X.sup.3-PBIGVRATB; X.sup.1-NYATGNLPG-X.sup.3-GCSFSIFLLAL; X.sup.1-SRNLGKVIDTLTC-X.sup.3-LMGYIPLVGA; X.sup.1-GYIPLVGAPL-X.sup.3-VARALAHGVRV; X.sup.1-GGGQIVGGVYLLP-X.sup.3-PRLGVRATR; X.sup.1-LTFLVRSVLL.sup.1-X.sup.3-GSVLIVRGSLVH; X.sup.1-TAYERNleCNIL-X.sup.3-GRFQTVVQBA; X.sup.1-SDPLVVAASIV-X.sup.3-ASIVGILHLIL; X.sup.1-LIFLARSALIL-X.sup.3-SALILRGSVAH; X.sup.1-IAYERMCNIL-X.sup.3-GKFQTAAQRA; and X.sup.1-LEPLVIAGILA-X.sup.3-GSLVGLLHIVL; X.sup.1-NLVPMVATV-X.sup.3-NLVPMATV; X.sup.1-GYLPAVGAPIG-X.sup.3-VIRVIAHGLRL; X.sup.1-IRNLGRVIETLTG-X.sup.3-LNleGYIPLIGA; X.sup.1-GVYDFAFRDLC-X.sup.3-GFAFRDLCIVYR, X.sup.1-GVFDYAFRDIN-X.sup.3-GFAYRDINLAYR, X.sup.1-GATPVDLLGA-X.sup.3-GALNLCLPMR, X.sup.1-GVTPAGLIGV-X.sup.3-GALQIBLPLR, and X.sup.1-IRNLGRVIETLTLNleGYIPLIGA-X.sup.3-IRNLGRVIETLTLNleGYIPLIGA; optionally with an X.sup.5 in the C-terminal of the peptide; wherein X.sup.1 and X.sup.3 and X.sup.5 refers to X.sup.1, X.sup.3, and X.sup.5 of formula II.
61. An isolated multimeric, such as dimeric peptide comprising two or more monomeric peptides, each monomeric peptide independently comprising the following structure
(Z.sup.1-Z.sup.2).sub.1-Z.sup.3-(Z.sup.4-Z.sup.5).sub.2-Z.sup.6-(Z.sup.7-Z.sup.8).sub.3-Z.sup.9-(Z.sup.10-Z.sup.11).sub.4-Z.sup.12
wherein Z.sup.1, Z.sup.4, and optional Z.sup.7 and Z.sup.10 defines a linear sequence of one, two, or three arginine residues or derivatives thereof optionally followed by a glycine (G) or an alanine (A); Z.sup.2, Z.sup.5, Z.sup.8 and Z.sup.11 defines an optional amino acid selected from cysteine (C), lysine (K), aspartic acid (D), asparagine (N), glutamic acid (E), glutamine (Q), 2,3-Diaminopropionic acid (Dpr), tryptophan (W), or tyrosine (Y) or a derivative thereof; Z.sup.3, and optional Z.sup.6, Z.sup.9 and Z.sup.12 defines any chemical moiety, such as a linear amino acid sequence, said monomeric peptides being covalently joined by one or more intermolecular bond.
62. The isolated multimeric, such as dimeric peptide according to embodiment 61, wherein two or more monomeric peptides are identical in sequence.
63. The isolated multimeric, such as dimeric peptide according to embodiment 61, wherein two or more monomeric peptides are different in sequence.
64. The isolated multimeric, such as dimeric peptide according to any of embodiments 61-63, comprising at least two peptides monomers, each peptide monomer independently being as defined in any one of embodiments 1-58.
65. The isolated multimeric, such as dimeric peptide according to any one of embodiments 61-64, wherein one or more peptide strands of the multimeric, such as dimeric peptide has delayed proteolytic degradation in the N-terminal, such as by incorporation of the first 1, 2, or 3 amino acids in the N-terminal in the D-form, or by incorporation of the first 1, 2, or 3 amino acids in the N-terminal in beta or gamma form.
66. The isolated multimeric, such as dimeric peptide according to any one of embodiments 61-65, which multimeric, such as dimeric peptide contain a helper epitope of at least 12 amino acids, such as at least 13, 14, 15 or 17 amino acids, which helper epitope consist of a combined sequence of amino acids, which is a sequence of amino acids from a first specific continuous antigenic peptide sequences, and a sequence of amino acids from at least one second specific continuous antigenic peptide sequence of the same or different protein derived from the same virus, any different virus, or any disease antigen, such as between 2-12 amino acids from the first specific continuous antigenic peptide sequences and 2-12 amino acids from the at least one second specific continuous antigenic antigenic peptide sequence.
67. The isolated multimeric, such as dimeric peptide according to any one of embodiments 61-66, wherein said intermolecular bond is a disulfide (SS) bond between two Cys residues.
68. The isolated multimeric, such as dimeric peptide according to any one of embodiments 61-67, wherein said intermolecular bond is a thioether bond between a Cys residue in the first monomeric peptide and a modified Lys residue in the at least one second monomeric peptide.
69. The isolated multimeric, such as dimeric peptide according to any one of embodiments 61-68, wherein said intermolecular bond is an oxime bond between a derivatized Lys residue in the first monomeric peptide and a derivatized Ser residue in the at least one second monomeric peptide.
70. The isolated multimeric, such as dimeric peptide according to any one of embodiments 61-69, wherein said intermolecular bond is a peptide bond between a N-methylated Lys side-chain in the first monomeric peptide and the side-chain of an Asp or Glu residue in the at least one second monomeric peptide.
71. The isolated multimeric, such as dimeric peptide according to any one of embodiments 61-70, wherein said intermolecular bond is an oxime bond between an aldehyde moiety, produced by oxidation of a serine residue in the first monomeric peptide and a free aminooxy group of a modified amino acid (aminooxy acid), such as derivatized diaminopropionic acid, Lysine or Ornithine in in the second monomeric peptide
72. The isolated multimeric, such as dimeric peptide according to any one of embodiments 61-71, wherein said monomeric peptides are linked by a polyethylene glycol (PEG) linker, such as through an Asp or a Glu residue in the first monomeric peptide and an Asp or a Glu residue in the at least one second monomeric peptide, or by a polyLys core.
73. The isolated multimeric, such as dimeric peptide according to any one of embodiments 61-72, wherein a C residue in Z.sup.2 of the first peptide monomer is linked to an amino acid selected from a K or a C residue in Z.sup.2 of the second monomer.
74. The isolated multimeric, such as dimeric peptide according to any one of embodiments 61-73, wherein a K residue in Z.sup.2 of the first peptide monomer is linked to an amino acid selected from a C, D or E residue in Z.sup.2 of the second monomer.
75. The isolated multimeric, such as dimeric peptide according to any one of embodiments 61-74, wherein a D residue in Z.sup.2 of the first peptide monomer is linked to an amino acid selected from a N or Q residue in Z.sup.2 of the second monomer.
76. The isolated multimeric, such as dimeric peptide according to any one of embodiments 61-75, wherein a E residue in Z.sup.2 of the first peptide monomer is linked to an amino acid selected from a N or Q residue in Z.sup.2 of the second monomer.
77. The isolated multimeric, such as dimeric peptide according to any one of embodiments 61-76, wherein a N residue in Z.sup.2 of the first peptide monomer is linked to a D or E residue in Z.sup.2 of the second monomer.
78. The isolated multimeric, such as dimeric peptide according to any one of embodiments 61-77, wherein a Q residue in Z.sup.2 of the first peptide monomer is linked to a D or E residue in Z.sup.2 of the second monomer.
79. The isolated multimeric, such as dimeric peptide according to any one of embodiments 61-78, wherein a Dpr(Aao) residue in Z.sup.2 of the first peptide monomer is linked to an Dpr(Ser) residue in Z.sup.2 of the second monomer.
80. The isolated multimeric, such as dimeric peptide according to any one of embodiments 61-79, wherein a W residue in Z.sup.2 of the first Z.sup.1-Z.sup.2 peptide repeat is linked to an Y residue in Z.sup.2 of the second Z.sup.1-Z.sup.2 peptide repeat.
81. The isolated multimeric, such as dimeric peptide according to any one of embodiments 61-80, wherein a Y residue in Z.sup.2 of the first Z.sup.1-Z.sup.2 peptide repeat is linked to an W residue in Z.sup.2 of the second Z.sup.1-Z.sup.2 peptide repeat.
82. Composition comprising two or more compounds selected from a monomeric peptide is as defined in any one of embodiments 1-60, and an isolated multimeric, such as dimeric peptide as defined in any one of embodiments 61-81.
83. Use of a peptide selected from a monomeric peptide is as defined in any one of embodiments 1-60, and an isolated multimeric, such as dimeric peptide as defined in any one of embodiments 61-81 for inducing an immune response in a subject, such as a humoral or Cell Mediated Immune (CMI) response.
84. An isolated nucleic acid or polynucleotide encoding a peptide according to any one of embodiments 1-61.
85. A vector comprising the nucleic acid or polynucleotide according to embodiment 84.
86. A host cell comprising the vector according to embodiment 85.
87. An immunogenic composition comprising at least one monomeric peptide according to any one of embodiments 1-61, an isolated multimeric, such as dimeric peptide according to any one of embodiments 61-81, a peptide composition according to embodiment 82, the nucleic acid or polynucleotide according to embodiment 84, or the vector according to embodiment 85; in combination with a pharmaceutically acceptable diluent or vehicle and optionally an immunological adjuvant.
88. The immunogenic composition according to embodiment 87 in the form of a vaccine composition.
89. A method for inducing an immune response in a subject against an antigen which comprises administration of at least one monomeric peptide according to any one of embodiments 1-60, an isolated multimeric, such as dimeric peptide according to any one of embodiments 61-79, a peptide composition according to embodiment 82, the nucleic acid or polynucleotide according to embodiment 84, or the vector according to embodiment 85; or the composition according to any one of embodiments 87-88.
90. A method for reducing and/or delaying the pathological effects of a disease antigen, such as an infectious agent in a subject infected with said agent or having said disease caused by said antigen, the method comprising administering an effective amount of at least one monomeric peptide according to any one of embodiments 1-60, an isolated multimeric, such as dimeric peptide according to any one of embodiments 61-81, a peptide composition according to embodiment 82, the nucleic acid or polynucleotide according to embodiment 84, or the vector according to embodiment 85; or the composition according to any one of embodiments 87-88.
91. A peptide according to any one of embodiments 1-81 for use as a medicament.
92. A peptide according to any one of embodiments 1-81 for treating the pathological effects of a disease antigen, such as an infectious agent in a subject infected with said agent or having said disease caused by said antigen.
93. A peptide according to any one of embodiments 1-81 for use in an in vitro assay, such as an ELISA assay, such as for diagnostic purposes.
94. Use of a peptide according to any one of embodiments 1-81 for in vitro assay, such as an ELISA assay, such as for diagnostic purposes.

(251) Sequence list (amino acids in bold represents suitable antigenic sequences that may be used as any of Z.sup.3, and optional Z.sup.6, Z.sup.9 and Z.sup.12 as defined in formula I of the present invention)

(252) TABLE-US-00022 SEQIDNO:1: AccessionnoAF009606;HepatitisCvirussubtype ispolyproteingene,completecds. MSTNPKPQRKTKRNTNRRPQDVKFPGGGQIVGGVYLLPRRGPRL GVRATRKTSERSQPRGRRQPIPKARRPEGRTWAQPGYPWPLYGNEGCGWAGWLLSPRG SRPSWGPTDPRRRSRNLGKVIDTLTCGFADLMGYIPLVGAPLGGAARALAHGVRVLED GVNYATGNLPGCSFSIFLLALLSCLTVPASAYQVRNSSGLYHVTNDCPNSSIVYEAAD AILHTPGCVPCVREGNASRCWVAVTPTVATRDGKLPTTQLRRHIDLLVGSATLCSALY VGDLCGSVFLVGQLFTFSPRRHWTTQDCNCSIYPGHITGHRMAWDMMMNWSPTAALVV AQLLRIPQAIMDMIAGAHWGVLAGIAYFSMVGNWAKVLVVLLLFAGVDAETHVTGGSA GRTTAGLVGLLTPGAKQNIQLINTNGSWHINSTALNCNESLNTGWLAGLFYQHKFNSS GCPERLASCRRLTDFAQGWGPISYANGSGLDERPYCWHYPPRPCGIVPAKSVCGPVYC FTPSPVVVGTTDRSGAPTYSWGANDTDVFVLNNTRPPLGNWFGCTWMNSTGFTKVCGA PPCVIGGVGNNTLLCPTDCFRKHPEATYSRCGSGPWITPRCMVDYPYRLWHYPCTINY TIFKVRMYVGGVEHRLEAACNWTRGERCDLEDRDRSELSPLLLSTTQWQVLPCSFTTL PALSTGLIHLHQNIVDVQYLYGVGSSIASWAIKWEYVVLLFLLLADARVCSCLWMMLL ISQAEAALENLVILNAASLAGTHGLVSFLVFFCFAWYLKGRWVPGAVYAFYGMWPLLL LLLALPQRAYALDTEVAASCGGVVLVGLMALTLSPYYKRYISWCMWWLQYFLTRVEAQ LHVWVPPLNVRGGRDAVILLMCVVHPTLVFDITKLLLAIFGPLWILQASLLKVPYFVR VQGLLRICALARKIAGGHYVQMAIIKLGALTGTYVYNHLTPLRDWAHNGLRDLAVAVE PVVFSRMETKLITWGADTAACGDIINGLPVSARRGQEILLGPADGMVSKGWRLLAPIT AYAQQTRGLLGCIITSLTGRDKNQVEGEVQIVSTATQTFLATCINGVCWTVYHGAGTR TIASPKGPVIQMYTNVDQDLVGWPAPQGSRSLTPCTCGSSDLYLVTRHADVIPVRRRG DSRGSLLSPRPISYLKGSSGGPLLCPAGHAVGLFRAAVCTRGVAKAVDFIPVENLETT MRSPVFTDNSSPPAVPQSFQVAHLHAPTGSGKSTKVPAAYAAQGYKVLVLNPSVAATL GFGAYMSKAHGVDPNIRTGVRTITTGSPITYSTYGKFLADGGCSGGAYDIIICDECHS TDATSILGIGTVLDQAETAGARLVVLATATPPGSVTVSHPNIEEVALSTTGEIPFYGK AIPLEVIKGGRHLIFCHSKKKCDELAAKLVALGINAVAYYRGLDVSVIPTSGDVVVVS TDALMTGFTGDFDSVIDCNTCVTQTVDFSLDPTFTIETTTLPQDAVSRTQRRGRTGRG KPGIYRFVAPGERPSGMFDSSVLCECYDAGCAWYELTPAETTVRLRAYMNTPGLPVCQ DHLEFWEGVFTGLTHIDAHFLSQTKQSGENFPYLVAYQATVCARAQAPPPSWDQMWKC LIRLKPTLHGPTPLLYRLGAVQNEVTLTHPITKYIMTCMSADLEVVTSTWVLVGGVLA ALAAYCLSTGCVVIVGRIVLSGKPAIIPDREVLYQEFDEMEECSQHLPYIEQGMMLAE QFKQKALGLLQTASRQAEVITPAVQTNWQKLEVFWAKHMWNFISGIQYLAGLSTLPGN PAIASLMAFTAAVTSPLTTGQTLLFNILGGWVAAQLAAPGAATAFVGAGLAGAAIGSV GLGKVLVDILAGYGAGVAGALVAFKIMSGEVPSTEDLVNLLPAILSPGALVVGVVCAA ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYVPESDAAARVTAILSSLTVTQLLR RLHQWISSECTTPCSGSWLRDIWDWICEVLSDFKTWLKAKLMPQLPGIPFVSCQRGYR GVWRGDGIMHTRCHCGAEITGHVKNGTMRIVGPRTCRNMWSGTFPINAYTTGPCTPLP APNYKFALWRVSAEEYVEIRRVGDFHYVSGMTTDNLKCPCQIPSPEFFTELDGVRLHR FAPPCKPLLREEVSFRVGLHEYPVGSQLPCEPEPDVAVLTSMLTDPSHITAEAAGRRL ARGSPPSMASSSASQLSAPSLKATCTANHDSPDAELIEANLLWRQEMGGNITRVESEN KVVILDSFDPLVAEEDEREVSVPAEILRKSRRFARALPVWARPDYNPPLVETWKKPDY EPPVVHGCPLPPPRSPPVPPPRKKRTVVLTESTLSTALAELATKSFGSSSTSGITGDN TTTSSEPAPSGCPPDSDVESYSSMPPLEGEPGDPDLSDGSWSTVSSGADTEDVVCCSM SYSWTGALVTPCAAEEQKLPINALSNSLLRHHNLVYSTTSRSACQRQKKVTFDRLQVL DSHYQDVLKEVKAAASKVKANLLSVEEACSLTPPHSAKSKFGYGAKDVRCHARKAVAH INSVWKDLLEDSVTPIDTTIMAKNEVFCVQPEKGGRKPARLIVFPDLGVRVCEKMALY DVVSKLPLAVMGSSYGFQYSPGQRVEFLVQAWKSKKTPMGFSYDTRCFDSTVTESDIR TEEAIYQCCDLDPQARVAIKSLTERLYVGGPLTNSRGENCGYRRCRASGVLTTSCGNT LTCYIKARAACRAAGLQDCTMLVCGDDLVVICESAGVQEDAASLRAFTEAMTRYSAPP GDPPQPEYDLELITSCSSNVSVAHDGAGKRVYYLTRDPTTPLARAAWETARHTPVNSW LGNIIMFAPTLWARMILMTHFFSVLIARDQLEQALNCEIYGACYSIEPLDLPPIIQRL HGLSAFSLHSYSPGEINRVAACLRKLGVPPLRAWRHRARSVRARLLSRGGRAAICGKY LFNWAVRTKLKLTPIAAAGRLDLSGWFTAGYSGGDIYHSVSHARPRWFWFCLLLLAAG VGIYLLPNR SEQIDNO:2: HCVcoreprotein,H77,AccessionAF009606 Genbanknumber:2316097 >gi|2316098|gb|AAB66324.1| polyprotein[HepatitisCvirus subtype1a] MSTNPKPQRKTKRNTNRRPQDVKFPGGGQIVGGVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARR PEGRTWAQPGYPWPLYGNEGCGWAGWLLSPRGSRPSWGPTDPRRRSRNLGKVIDTLTCGFADLMGYIPLVGAPLGGAAR ALAHGVRVLEDGVNYATGNLPGCSFSIFLLALLSCLTVPASA SEQIDNO:3: HepatitisCvirusmRNA,completecds; ACCESSIONM96362M72423; HepatitisCvirussubtype1b MSTNPKPQRKTKRNTNRRPQDIKFPGGGQIVGGVYLLPRRGPRL GVRATRKTSERSQPRGRRQPIPKARRPEGRAWAQPGYPWPLYGNEGLGWAGWLLSPRG SRPSWGPTDPRRKSRNLGKVIDTLTCGFADLMGYIPLVGAPLGGVARALAHGVRVLED GVNYATGNLPGCSFSIFLLALLSCLTTPVSAYEVRNASGMYHVTNDCSNSSIVYEAAD MIMHTPGCVPCVREDNSSRCWVALTPTLAARNASVPTTTLRRHVDLLVGVAAFCSAMY VGDLCGSVFLVSQLFTESPRRHETVQDCNCSIYPGRVSGHRMAWDMMMNWSPTTALVV SQLLRIPQAVVDMVTGSHWGILAGLAYYSMVGNWAKVLIAMLLFAGVDGTTHVTGGAQ GRAASSLTSLFSPGPVQHLQLINTNGSWHINRTALSCNDSLNTGFVAALFYKYRFNAS GCPERLATCRPIDTFAQGWGPITYTEPHDLDQRPYCWHYAPQPCGIVPTLQVCGPVYC FTPSPVAVGTTDRFGAPTYRWGANETDVLLLNNAGPPQGNWFGCTWMNGTGFTKTCGG PPCNIGGVGNNTLTCPTDCFRKHPGATYTKCGSGPWLTPRCLVDYPYRLWHYPCTVNF TIFKVRMYVGGAEHRLDAACNWTRGERCDLEDRDRSELSPLLLSTTEWQVLPCSFTTL PALSTGLIHLHQNIVDIQYLYGIGSAVVSFAIKWEYIVLLFLLLADARVCACLWMMLL VAQAEAALENLVVLNAASVAGAHGILSFIVFFCAAWYIKGRLVPGAAYALYGVWPLLL LLLALPPRAYAMDREMAASCGGAVFVGLVLLTLSPHYKVFLARFIWWLQYLITRTEAH LQVWVPPLNVRGGRDAIILLTCVVHPELIFDITKYLLAIFGPLMVLQAGITRVPYFVR AQGLIRACMLARKVVGGHYVQMVFMKLAALAGTYVYDHLTPLRDWAHTGLRDLAVAVE PVVFSDMETKVITWGADTAACGDIILALPASARRGKEILLGPADSLEGQGWRLLAPIT AYSQQTRGLLGCIITSLTGRDKNQVEGEVQVVSTATQSFLATCINGVCWTVFHGAGSK TLAGPKGPITQMYTNVDQDLVGWPAPPGARSLTPCTCGSSDLYLVTRHADVIPVRRRG DGRGSLLPPRPVSYLKGSSGGPLLCPSGHAVGILPAAVCTRGVAMAVEFIPVESMETT MRSPVFTDNPSPPAVPQTFQVAHLHAPTGSGKSTRVPAAYAAQGYKVLVLNPSVAATL GFGAYMSKAHGIDPNLRTGVRTITTGAPITYSTYGKFLADGGGSGGAYDIIMCDECHS TDSTTIYGIGTVLDQAETAGARLVVLSTATPPGSVTVPHLNIEEVALSNTGEIPFYGK AIPIEAIKGGRHLIFCHSKKKCDELAAKLSGLGLNAVAYYRGLDVSVIPTSGDVVVVA TDALMTGFTGDFDSVIDCNTCVTQTVDFSLDPTFTIETTTVPQDAVSRSQRRGRTGRG RAGIYRFVTPGERPSGMFDSSVLCECYDAGCAWYELTPAETSVRLRAYLNTPGLPVCQ DHLEFSEGVFTGLTHIDAHFLSQTKQAGENFPYLVAYQATVCARAQAPPPSWDEMWRC LIRLKPTLHGPTPLLYRLGAVQNEVTLTHPITKFIMTCMSADLEVVTSTWVLVGGVLA ALAAYCLTTGSVVIVGRIILSGKPAIIPDREVLYQEFDEMEECASHLPYFEQGMQLAE QFKQKALGLLQTATKQAEAAAPVVESKWRALETFWAKHMWNFISGIQYLAGLSTLPGN PAIRSPMAFTASITSPLTTQHTLLFNILGGWVAAQLAPPSAASAFVGAGIAGAAVGTI GLGKVLVDILAGYGAGVAGALVAFKIMSGEMPSAEDMVNLLPAILSPGALVVGIVCAA ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPRHYVPESEPAARVTQILSSLTITQLLK RLHQWINEDCSTPCSSSWLREIWDWICTVLTDFKTWLQSKLLPRLPGVPFFSCQRGYK GVWRGDGIMHTTCPCGAQITGHVKNGSMRIVGPKTCSNTWYGTFPINAYTTGPCTPSP APNYSKALWRVAAEEYVEVTRVGDFHYVTGMTTDNVKCPCQVPAPEFFTEVDGVRLHR YAPACRPLLREEVVFQVGLHQYLVGSQLPCEPEPDVAVLTSMLTDPSHITAETAKRRL ARGSPPSLASSSASQLSAPSLKATCTTHHDSPDADLIEANLLWRQEMGGNITRVESEN KVVILDSFDPLRAEDDEGEISVPAEILRKSRKFPPALPIWAPPDYNPPLLESWKDPDY VPPVVHGCPLPPTKAPPIPPPRRKRTVVLTESTVSSALAELATKTFGSSGSSAIDSGT ATAPPDQASGDGDRESDVESFSSMPPLEGEPGDPDLSDGSWSTVSEEASEDVVCCSMS YTWTGALITPCAAEESKLPINPLSNSLLRHHNMVYATTSRSAGLRQKKVTFDRLQVLD DHYRDVLKEMKAKASTVKAKLLSVEEACKLTPPHSAKSKFGYGAKDVRSLSSRAVTHI RSVWKDLLEDTETPISTTIMAKNEVFCVQPEKGGRKPARLIVFPDLGVRVCEKMALYD VVSTLPQAVMGSSYGFQYSPKQRVEFLVNTWKSKKCPMGFSYDTRCFDSTVTENDIRV EESIYQCCDLAPEAKLAIKSLTERLYIGGPLTNSKGQNCGYRRCRASGVLTTSCGNTL TCYLKATAACRAAKLRDCTMLVNGDDLVVICESAGTQEDAASLRVFTEAMTRYSAPPG DPPQPEYDLELITSCSSNVSVAHDASGKRVYYLTRDPTTPLARAAWETARHTPVNSWL GNIIMYAPTLWARMILMTHFFSILLAQEQLEKTLDCQIYGACYSIEPLDLPQIIERLH GLSAFSLHSYSPGEINRVASCLRKLGVPPLRAWRHRARSVRAKLLSQGGRAATCGKYL FNWAVRTKLKLTPIPAASRLDLSGWFVAGYSGGDIYHSLSRARPRWFMLCLLLLSVGV GIYLLPNR SEQIDNO:4, nucleocapsidproteinofinfluenzaAvirus 1MASQGTKRSYEQMETSGERQNATEIRASVGRMVGGIGRFYIQMCTELKLSDHEGRLIQNS 61ITIERMVLSAFDERRNKYLEEHPSAGKDPKKTGGPIYRRRDGKWMRELILYDKEEIRRIW 121RQANNGEDATAGLTHMMIWHSNLNDATYQRTRALVRTGMDPRMCSLMQGSTLPRRSGAAG 181AAVKGVGTMVMELIRMIKRGINDRNFWRGENGRRTRIAYERMCNILKGKFQTAAQRAMMD 241QVRESRNPGNAEIEDLIFLARSALILRGSVAHKSCLPACVYGLAVASGYDFEREGYSLVG 301IDPFRLLQNSQVFSLIRPNENPAHKSQLVWMACHSAAFEDLRVSSFIRGTRVVPRGQLST 361RGVQIASNENMETMDSSTLELRSRYWAIRTRSGGNTNQQRASAGQISVQPTFSVQRNLPF 421ERATIMAAFTGNTEGRTSDMRTEIIRMMENARPEDVSFQGRGVFELSDEKATNPIVPSFD 481MSNEGS SEQIDNO:5 >gi|73919153|ref|YP_308840.1| matrixprotein2 [InfluenzaAvirus(A/NewYork/392/2004(H3N2))] MSLLTEVETPIRNEWGCRCNDSSDPLVVAASIIGILHLILWILDRLFFKCVYRLEKHGLKRGPSTEGVPE 70 SMREEYRKEQQNAVDADDSHFVSIELE SEQIDNO:6 >gi|73919147|ref|YP_308843.1| nucleocapsidprotein [InfluenzaAvirus(A/NewYork/392/2004(H3N2))] MASQGTKRSYEQMETDGDRQNATEIRASVGKMIDGIGRFYIQMCTELKLSDHEGRLIQNSLTIEKMVLSA 70 FDERRNKYLEEHPSAGKDPKKTGGPIYRRVDGKWMRELVLYDKEEIRRIWRQANNGEDATAGLTHIMIWH 140 SNLNDATYQRTRALVRTGMDPRMCSLMQGSTLPRRSGAAGAAVKGIGTMVMELIRMVKRGINDRNFWRGE 210 NGRKTRSAYERMCNILKGKFQTAAQRAMVDQVRESRNPGNAEIEDLIFLARSALILRGSVAHKSCLPACA 280 YGPAVSSGYDFEKEGYSLVGIDPFKLLQNSQIYSLIRPNENPAHKSQLVWMACHSAAFEDLRLLSFIRGT 350 KVSPRGKLSTRGVQIASNENMDNMGSSTLELRSGYWAIRTRSGGNTNQQRASAGQTSVQPTFSVQRNLPF 420 EKSTIMAAFTGNTEGRTSDMRAEIIRMMEGAKPEEVSFRGRGVFELSDEKATNPIVPSFDMSNEGSYFFG 490 DNAEEYDN -- SEQIDNO:7 >gi|56583270|ref|NP_040979.2| matrixprotein2 [InfluenzaAvirus(A/PuertoRico/8/34(H1N1))] MSLLTEVETPIRNEWGCRCNGSSDPLAIAANIIGILHLILWILDRLFFKCIYRREKYGLKGGPSTEGVPK SMREEYRKEQQSAVDADDGHFVSIELE SEQIDNO:8 >gi|8486130|ref|NP_040982.1| nucleocapsidprotein [InfluenzaAvirus(A/PuertoRico/8/34(H1N1))] MASQGTKRSYEQMETDGERQNATEIRASVGKMIGGIGRFYIQMCTELKLSDYEGRLIQNSLTIERMVLSA FDERRNKYLEEHPSAGKDPKKTGGPIYRRVNGKWMRELILYDKEEIRRIWRQANNGDDATAGLTHMMIWH SNLNDATYQRTRALVRTGMDPRMCSLMQGSTLPRRSGAAGAAVKGVGTMVMELVRMIKRGINDRNFWRGE NGRKTRIAYERMCNILKGKFQTAAQKAMMDQVRESRDPGNAEFEDLTFLARSALILRGSVAHKSCLPACV YGPAVASGYDFEREGYSLVGIDPFRLLQNSQVYSLIRPNENPAHKSQLVWMACHSAAFEDLRVLSFIKGT KVVPRGKLSTRGVQIASNENMETMESSTLELRSRYWAIRTRSGGNTNQQRASAGQISIQPTFSVQRNLPF DRTTVMAAFTGNTEGRTSDMRTEIIRMMESARPEDVSFQGRGVFELSDEKAASPIVPSFDMSNEGSYFFG DNAEEYDN -- SEQIDNO:9 >gi|73912687|ref|YP_308853.1| membraneproteinM2 [InfluenzaAvirus(A/Korea/426/68(H2N2))] MSLLTEVETPIRNEWGCRCNDSSDPLVVAASIIGILHFILWILDRLFFKCIYRFEKHGLKRGPSTEGVPE SMREEYRKEQQSAVDADDSHFVSIELE SEQIDNO:10 >gi|73921307|ref|YP_308871.1| nucleoprotein[InfluenzaAvirus (A/Korea/426/68(H2N2))] MASQGTKRSYEQMETDGERQNATEIRASVGKMIDGIGRFYIQMCTELKLSDYEGRLIQNSLTIERMVLSA FDERRNKYLEEHPSAGKDPKKTGGPIYKRVDGKWMRELVLYDKEEIRRIWRQANNGDDATAGLTHMMIWH SNLNDTTYQRTRALVRTGMDPRMCSLMQGSTLPRRSGAAGAAVKGVGTMVMELIRMIKRGINDRNFWRGE NGRKTRSAYERMCNILKGKFQTAAQRAMMDQVRESRNPGNAEIEDLIFLARSALILRGSVAHKSCLPACV YGPAIASGYNFEKEGYSLVGIDPFKLLQNSQVYSLIRPNENPAHKSQLVWMACNSAAFEDLRVLSFIRGT KVSPRGKLSTRGVQIASNENMDTMESSTLELRSRYWAIRTRSGGNTNQQRASAGQISVQPAFSVQRNLPF DKPTIMAAFTGNTEGRTSDMRAEIIRMMEGAKPEEMSFQGRGVFELSDEKATNPIVPSFDMSNEGSYFFG DNAEEYDN SEQIDNO:11 >gi|330647|gb|AAA45994.1| pp65[Humanherpesvirus5] MASVLGPISGHVLKAVFSRGDTPVLPHETRLLQTGIHVRVSQPSLILVSQYTPDSTPCHRGDNQLQVQHT 70 YFTGSEVENVSVNVHNPTGRSICPSQEPMSIYVYALPLKMLNIPSINVHHYPSAAERKHRHLPVADAVIH 140 ASGKQMWQARLTVSGLAWTRQQNQWKEPDVYYTSAFVFPTKDVALRHVVCAHELVCSMENTRATKMQVIG 210 DQYVKVYLESFCEDVPSGKLFMHVTLGSDVEEDLTMTRNPQPFMRPHERNGFTVLCPKNMIIKPGKISHI 280 MLDVAFTSHEHFGLLCPKSIPGLSISGNLLMNGQQIFLEVQAIRETVELRQYDPVAALFFFDIDLLLQRG 350 PQYSEHPTFTSQYRIQGKLEYRHTWDRHDEGAAQGDDDVWTSGSDSDEELVTTERKTPRVTGGGAMAGAS 420 TSAGRKRKSASSATACTAGVMTRGRLKAESTVAPEEDTDEDSDNEIHNPAVFTWPPWQAGILARNLVPMV 490 ATVQGQNLKYQEFFWDANDIYRIFAELEGVWQPAAQPKRRRHRQDALPGPCIASTPKKHRG 541 SEQIDNO:12 >gi|33330937|gb|AAQ10712.1| putativetransformingproteinE6 [Humanpapillomavirustype16] MHQKRTAMFQDPQERPGKLPQLCTELQTTIHDIILECVYCKQQLLRREVYDFAFRDLCIVYRDGNPYAVC 70 DKCLKFYSKISEYRHYCYSVYGTTLEQQYNKPLCDLLIRCINCQKPLCPEEKQRHLDKKQRFHNIRGRWT 140 GRCMSCCRSSRTRRETQL SEQIDNO:13 >gi|56583270|ref|NP_040979.2| matrixprotein2 [InfluenzaAvirus(A/PuertoRico/8/34(H1N1))] MSLLTEVETPIRNEWGCRCNGSSDPLAIAANIIGILHLILWILDRLFFKCIYRRFKYGLKGGPSTEGVPK SMREEYRKEQQSAVDADDGHFVSIELE SEQIDNO:14 >gi|8486139|ref|NP_040987.1| PB2protein [InfluenzaAvirus(A/PuertoRico/8/34(H1N1))] MERIKELRNLMSQSRTREILTKTTVDHMAIIKKYTSGRQEKNPALRMKWMMAMKYPITADKRITEMIPER NEQGQTLWSKMNDAGSDRVMVSPLAVTWWNRNGPMTNTVHYPKIYKTYFERVERLKHGTFGPVHFRNQVK IRRRVDINPGHADLSAKEAQDVIMEVVFPNEVGARILTSESQLTITKEKKEELQDCKISPLMVAYMLERE LVRKTRFLPVAGGTSSVYIEVLHLTQGTCWEQMYTPGGEVKNDDVDQSLIIAARNIVRRAAVSADPLASL LEMCHSTQIGGIRMVDILKQNPTEEQAVGICKAAMGLRISSSFSFGGFTFKRTSGSSVKREEEVLTGNLQ TLKIRVHEGYEEFTMVGRRATAILRKATRRLIQLIVSGRDEQSIAEAIIVAMVFSQEDCMIKAVRGDLNF VNRANQRLNPMHQLLRHFQKDAKVLFQNWGVEPIDNVMGMIGILPDMTPSIEMSMRGVRISKMGVDEYSS TERVVVSIDRFLRVRDQRGNVLLSPEEVSETQGTEKLTITYSSSMMWEINGPESVLVNTYQWIIRNWETV KIQWSQNPTMLYNKMEFEPFQSLVPKAIRGQYSGFVRTLFQQMRDVLGTFDTAQIIKLLPFAAAPPKQSR MQFSSFTVNVRGSGMRILVRGNSPVFNYNKATKRLTVLGKDAGTLTEDPDEGTAGVESAVLRGFLILGKE DRRYGPALSINELSNLAKGEKANVLIGQGDVVLVMKRKRDSSILTDSQTATKRIRMAIN SEQIDNO:15 >gi|8486137|ref|NP_040986.1| polymerasePA [InfluenzaAvirus(A/PuertoRico/8/34(H1N1))] MEDFVRQCFNPMIVELAEKTMKEYGEDLKIETNKFAAICTHLEVCFMYSDFHFINEQGESIIVELGDPNA LLKHRFEIIEGRDRTMAWTVVNSICNTTGAEKPKFLPDLYDYKENRFIEIGVTRREVHIYYLEKANKIKS EKTHIHIFSFTGEEMATKADYTLDEESRARIKTRLFTIRQEMASRGLWDSFRQSERGEETIEERFEITGT MRKLADQSLPPNFSSLENFRAYVDGFEPNGYIEGKLSQMSKEVNARIEPFLKTTPRPLRLPNGPPCSQRS KFLLMDALKLSIEDPSHEGEGIPLYDAIKCMRTFFGWKEPNVVKPHEKGINPNYLLSWKQVLAELQDIEN EEKIPKTKNMKKTSQLKWALGENMAPEKVDFDDCKDVGDLKQYDSDEPELRSLASWIQNEFNKACELTDS SWIELDEIGEDVAPIEHIASMRRNYFTSEVSHCRATEYIMKGVYINTALLNASCAAMDDFQLIPMISKCR TKEGRRKTNLYGFIIKGRSHLRNDTDVVNFVSMEFSLTDPRLEPHKWEKYCVLEIGDMLLRSAIGQVSRP MFLYVRTNGTSKIKMKWGMEMRRCLLQSLQQIESMIEAESSVKEKDMTKEFFENKSETWPIGESPKGVEE SSIGKVCRTLLAKSVFNSLYASPQLEGFSAESRKLLLIVQALRDNLEPGTFDLGGLYEAIEECLINDPWV LLNASWFNSFLTHALS SEQIDNO:16 >gi|8486133|ref|NP_040984.1| nonstructuralproteinNS1 [InfluenzaAvirus(A/PuertoRico/8/34(H1N1))] MDPNTVSSFQVDCFLWHVRKRVADQELGDAPFLDRLRRDQKSLRGRGSTLGLDIETATRAGKQIVERILK EESDEALKMTMASVPASRYLTDMTLEEMSREWSMLIPKQKVAGPLCIRMDQAIMDKNIILKANFSVIFDR LETLILLRAFTEEGAIVGEISPLPSLPGHTAEDVKNAVGVLIGGLEWNDNTVRVSETLQRFAWRSSNENG RPPLTPKQKREMAGTIRSEV SEQIDNO:17 >gi|8486132|ref|NP_040983.1| nonstructuralproteinNS2 [InfluenzaAvirus(A/PuertoRico/8/34(H1N1))] MDPNTVSSFQDILLRMSKMQLESSSEDLNGMITQFESLKLYRDSLGEAVMRMGDLHSLQNRNEKWREQLG QKFEEIRWLIEEVRHKLKVTENSFEQITFMQALHLLLEVEQEIRTFSFQLI SEQIDNO:18 >gi|8486128|ref|NP_040981.1| neuraminidase [InfluenzaAvirus(A/PuertoRico/8/34(H1N1))] MNPNQKIITIGSICLVVGLISLILQIGNIISIWISHSIQTGSQNHTGICNQNIITYKNSTWVKDTTSVIL TGNSSLCPIRGWAIYSKDNSIRIGSKGDVFVIREPFISCSHLECRTFFLTQGALLNDRHSNGTVKDRSPY RALMSCPVGEAPSPYNSRFESVAWSASACHDGMGWLTIGISGPDNGAVAVLKYNGIITETIKSWRKKILR TQESECACVNGSCFTIMTDGPSDGLASYKIFKIEKGKVTKSIELNAPNSHYEECSCYPDTGKVMCVCRDN WHGSNRPWVSFDQNLDYQIGYICSGVFGDNPRPKDGTGSCGPVYVDGANGVKGFSYRYGNGVWIGRTKSH SSRHGFEMIWDPNGWTETDSKFSVRQDVVAMTDWSGYSGSFVQHPELTGLDCIRPCFWVELIRGRPKEKT IWTSASSISFCGVNSDTVDWSWPDGAELPFTIDK SEQIDNO:19 >gi|8486126|ref|NP_040980.1| haemagglutinin[InfluenzaAvirus (A/PuertoRico/8/34(H1N1))] MKANLLVLLCALAAADADTICIGYHANNSTDTVDTVLEKNVTVTHSVNLLEDSHNGKLCRLKGIAPLQLG KCNIAGWLLGNPECDPLLPVRSWSYIVETPNSENGICYPGDFIDYEELREQLSSVSSFERFEIFPKESSW PNHNTTKGVTAACSHAGKSSFYRNLLWLTEKEGSYPKLKNSYVNKKGKEVLVLWGIHHPSNSKDQQNIYQ NENAYVSVVTSNYNRRFTPEIAERPKVRDQAGRMNYYWTLLKPGDTIIFEANGNLIAPRYAFALSRGFGS GIITSNASMHECNTKCQTPLGAINSSLPFQNIHPVTIGECPKYVRSAKLRMVTGLRNIPSIQSRGLFGAI AGFIEGGWTGMIDGWYGYHHQNEQGSGYAADQKSTQNAINGITNKVNSVIEKMNIQFTAVGKEFNKLEKR MENLNKKVDDGFLDIWTYNAELLVLLENERTLDFHDSNVKNLYEKVKSQLKNNAKEIGNGCFEFYHKCDN ECMESVRNGTYDYPKYSEESKLNREKVDGVKLESMGIYQILAIYSTVASSLVLLVSLGAISFWMCSNGSL QCRICI SEQIDNO:20 >gi|8486123|ref|NP_040978.1| matrixprotein1[InfluenzaAvirus (A/PuertoRico/8/34(H1N1))] MSLLTEVETYVLSIIPSGPLKAEIAQRLEDVFAGKNTDLEVLMEWLKTRPILSPLTKGILGFVFTLTVPS ERGLQRRRFVQNALNGNGDPNNMDKAVKLYRKLKREITFHGAKEISLSYSAGALASCMGLIYNRMGAVTT EVAFGLVCATCEQIADSQHRSHRQMVTTTNPLIRHENRMVLASTTAKAMEQMAGSSEQAAEAMEVASQAR QMVQAMRTIGTHPSSSAGLKNDLLENLQAYQKRMGVQMQRFK SEQIDNO:21 >gi|83031685|ref|YP_418248.1| PB1-F2protein[InfluenzaAvirus (A/PuertoRico/8/34(H1N1))] MGQEQDTPWILSTGHISTQKRQDGQQTPKLEHRNSTRLMGHCQKTMNQVVMPKQIVYWKQWLSLRNPILV FLKTRVLKRWRLFSKHE SEQIDNO:22 >gi|8486135|ref|NP_040985.1| polymerase1PB1[InfluenzaAvirus (A/PuertoRico/8/34(H1N1))] MDVNPTLLFLKVPAQNAISTTFPYTGDPPYSHGTGTGYTMDTVNRTHQYSEKARWTTNTETGAPQLNPID GPLPEDNEPSGYAQTDCVLEAMAFLEESHPGIFENSCIETMEVVQQTRVDKLTQGRQTYDWTLNRNQPAA TALANTIEVFRSNGLTANESGRLIDFLKDVMESMKKEEMGITTHFQRKRRVRDNMTKKMITQRTIGKRKQ RLNKRSYLIRALTLNTMTKDAERGKLKRRAIATPGMQIRGFVYFVETLARSICEKLEQSGLPVGGNEKKA KLANVVRKMMTNSQDTELSLTITGDNTKWNENQNPRMFLAMITYMTRNQPEWFRNVLSIAPIMFSNKMAR LGKGYMFESKSMKLRTQIPAEMLASIDLKYFNDSTRKKIEKIRPLLIEGTASLSPGMMMGMFNMLSTVLG VSILNLGQKRYTKTTYWWDGLQSSDDFALIVNAPNHEGIQAGVDRFYRTCKLHGINMSKKKSYINRTGTF EFTSFFYRYGFVANFSMELPSFGVSGSNESADMSIGVTVIKNNMINNDLGPATAQMALQLFIKDYRYTYR CHRGDTQIQTRRSFEIKKLWEQTRSKAGLLVSDGGPNLYNIRNLHIPEVCLKWELMDEDYQGRLCNPLNP FVSHKEIESMNNAVMMPAHGPAKNMEYDAVATTHSWIPKRNRSILNTSQRGVLEDEQMYQRCCNLFEKFF PSSSYRRPVGISSMVEAMVSRARIDARIDFESGRIKKEEFTEIMKICSTIEELRRQK SEQIDNO:23 >gi|8486130|ref|NP_040982.1| nucleocapsidprotein [InfluenzaAvirus(A/PuertoRico/8/34(H1N1))] MASQGTKRSYEQMETDGERQNATEIRASVGKMIGGIGRFYIQMCTELKLSDYEGRLIQNSLTIERMVLSA FDERRNKYLEEHPSAGKDPKKTGGPIYRRVNGKWMRELILYDKEEIRRIWRQANNGDDATAGLTHMMIWH SNLNDATYQRTRALVRTGMDPRMCSLMQGSTLPRRSGAAGAAVKGVGTMVMELVRMIKRGINDRNFWRGE NGRKTRIAYERMCNILKGKFQTAAQKAMMDQVRESRDPGNAEFEDLTFLARSALILRGSVAHKSCLPACV YGPAVASGYDFEREGYSLVGIDPFRLLQNSQVYSLIRPNENPAHKSQLVWMACHSAAFEDLRVLSFIKGT KVVPRGKLSTRGVQIASNENMETMESSTLELRSRYWAIRTRSGGNTNQQRASAGQISIQPTFSVQRNLPF DRTTVMAAFTGNTEGRTSDMRTEIIRMMESARPEDVSFQGRGVFELSDEKAASPIVPSFDMSNEGSYFFG DNAEEYDN SEQIDNO:24 >gi|73918826|ref|YP_308855.1| polymerase2[InfluenzaAvirus (A/Korea/426/1968(H2N2))] MERIKELRNLMSQSRTREILTKTTVDHMAIIKKYTSGRQEKNPSLRMKWMMAMKYPITADKRITEMVPER NEQGQTLWSKMSDAGSDRVMVSPLAVTWWNRNGPMTSTVHYPKIYKTYFEKVERLKHGTFGPVHFRNQVK IRRRVDINPGHADLSAKEAQDVIMEVVFPNEVGARILTSESQLTITKEKKEELQDCKISPLMVAYMLERE LVRKTRFLPVAGGTSSVYIEVLHLTQGTCWEQMYTPGGEVRNDDVDQSLIIAARNIVRRAAVSADPLASL LEMCHSTQIGGTRMVDILRQNPTEEQAVDICKAAMGLRISSSFSFGGFTFKRTSGSSIKREEEVLTGNLQ TLKIRVHEGYEEFTMVGKRATAILRKATRRLVQLIVSGRDEQSIAEAIIVAMVFSQEDCMIKAVRGDLNF VNRANQRLNPMHQLLRHFQKDAKVLFQNWGIEHIDNVMGMIGVLPDMTPSTEMSMRGIRVSKMGVDEYSS TERVVVSIDRFLRVRDQRGNVLLSPEEVSETQGTEKLTITYSSSMMWEINGPESVLVNTYQWIIRNWETV KIQWSQNPTMLYNKMEFEPFQSLVPKAIRGQYSGFVRTLFQQMRDVLGTFDTTQIIKLLPFAAAPPKQSR MQFSSLTVNVRGSGMRILVRGNSPVFNYNKTTKRLTILGKDAGTLTEDPDEGTSGVESAVLRGFLILGKE DRRYGPALSINELSTLAKGEKANVLIGQGDVVLVMKRKRDSSILTDSQTATKRIRMAIN SEQIDNO:25 >gi|73919145|ref|YP_308850.1| hemagglutinin[InfluenzaAvirus (A/Korea/426/68(H2N2))] MAIIYLILLFTAVRGDQICIGYHANNSTEKVDTILERNVTVTHAKDILEKTHNGKLCKLNGIPPLELGDC SIAGWLLGNPECDRLLSVPEWSYIMEKENPRYSLCYPGSFNDYEELKHLLSSVKHFEKVKILPKDRWTQH TTTGGSWACAVSGKPSFFRNMVWLTRKGSNYPVAKGSYNNTSGEQMLIIWGVHHPNDEAEQRALYQNVGT YVSVATSTLYKRSIPEIAARPKVNGLGRRMEFSWTLLDMWDTINFESTGNLVAPEYGFKISKRGSSGIMK TEGTLENCETKCQTPLGAINTTLPFHNVHPLTIGECPKYVKSEKLVLATGLRNVPQIESRGLFGAIAGFI EGGWQGMVDGWYGYHHSNDQGSGYAADKESTQKAFNGITNKVNSVIEKMNTQFEAVGKEFSNLEKRLENL NKKMEDGFLDVWTYNAELLVLMENERTLDFHDSNVKNLYDKVRMQLRDNVKELGNGCFEFYHKCDNECMD SVKNGTYDYPKYEEESKLNRNEIKGVKLSSMGVYQILAIYATVAGSLSLAIMMAGISFWMCSNGSLQCRI CI SEQIDNO:26 >gi|73912688|ref|YP_308854.1| membraneproteinM1 [InfluenzaAvirus(A/Korea/426/68(H2N2))] MSLLTEVETYVLSIVPSGPLKAEIAQRLEDVFAGKNTDLEALMEWLKTRPILSPLTKGILGFVFTLTVPS ERGLQRRRFVQNALNGNGDPNNMDRAVKLYRKLKREITFHGAKEVALSYSAGALASCMGLIYNRMGAVTT EVAFAVVCATCEQIADSQHRSHRQMVTTTNPLIRHENRMVLASTTAKAMEQMAGSSEQAAEAMEVASQAR QMVQAMRAIGTPPSSSAGLKDDLLENLQAYQKRMGVQMQRFK SEQIDNO:27 >gi|73912687|ref|YP_308853.1| membraneproteinM2 [InfluenzaAvirus(A/Korea/426/68(H2N2))] MSLLTEVETPIRNEWGCRCNDSSDPLVVAASIIGILHFILWILDRLFFKCIYRFFKHGLKRGPSTEGVPE SMREEYRKEQQSAVDADDSHFVSIELE SEQIDNO:28 >gi|73912685|ref|YP_308852.1| polymerasePA[InfluenzaAvirus (A/Korea/426/68(H2N2))] MEDFVRQCFNPMIVELAEKAMKEYGEDLKIETNKFAAICTHLEVCFMYSDFHFINEQGESIMVELDDPNA LLKHRFEIIEGRDRTMAWTVVNSICNTTGAEKPKFLPDLYDYKENRFIEIGVTRREVHIYYLEKANKIKS ENTHIHIFSFTGEEMATKADYTLDEESRARIKTRLFTIRQEMANRGLWDSFRQSERGEETIEERFEITGT MRRLADQSLPPNFSCLENFRAYVDGFEPNGYIEGKLSQMSKEVNAKIEPFLKTTPRPIRLPDGPPCFQRS KFLLMDALKLSIEDPSHEGEGIPLYDAIKCMRTFFGWKEPYIVKPHEKGINPNYLLSWKQVLAELQDIEN EEKIPRTKNMKKTSQLKWALGENMAPEKVDFDNCRDISDLKQYDSDEPELRSLSSWIQNEFNKACELTDS IWIELDEIGEDVAPIEHIASMRRNYFTAEVSHCRATEYIMKGVYINTALLNASCAAMDDFQLIPMISKCR TKEGRRKTNLYGFIIKGRSHLRNDTDVVNFVSMEFSLTDPRLEPHKWEKYCVLEIGDMLLRSAIGQMSRP MFLYVRTNGTSKIKMKWGMEMRPCLLQSLQQIESMVEAESSVKEKDMTKEFFENKSETWPIGESPKGVEE GSIGKVCRTLLAKSVFNSLYASPQLEGFSAESRKLLLVVQALRDNLEPGTFDLGGLYEAIEECLINDPWV LLNASWFNSFLTHALR SEQIDNO:29 >gi|73921833|ref|YP_308877.1| PB1-F2protein[InfluenzaAvirus (A/Korea/426/68(H2N2))] MGQEQDTPWTQSTEHINIQKRGSGQQTRKLERPNLTQLMDHYLRTMNQVDMHKQTASWKQWLSLRNHTQE SLKIRVLKRWKLFNKQEWTN SEQIDNO:30 >gi|73912683|ref|YP_308851.1| PB1polymerasesubunit [InfluenzaAvirus(A/Korea/426/68(H2N2))] MDVNPTLLFLKVPAQNAISTTFPYTGDPPYSHGTGTGYTMDTVNRTHQYSEKGKWTTNTETGAPQLNPID GPLPEDNEPSGYAQTDCVLEAMAFLEESHPGIFENSCLETMEVIQQTRVDKLTQGRQTYDWTLNRNQPAA TALANTIEVFRSNGLTANESGRLIDFLKDVIESMDKEEMEITTHFQRKRRVRDNMTKKMVTQRTIGKKKQ RLNKRSYLIRALTLNTMTKDAERGKLKRRAIATPGMQIRGFVHFVETLARNICEKLEQSGLPVGGNEKKA KLANVVRKMMTNSQDTELSFTITGDNTKWNENQNPRVFLAMITYITRNQPEWFRNVLSIAPIMFSNKMAR LGKGYMFESKSMKLRTQIPAEMLASIDLKYFNESTRKKIEKIRPLLIDGTVSLSPGMMMGMFNMLSTVLG VSILNLGQKKYTKTTYWWDGLQSSDDFALIVNAPNHEGIQAGVNRFYRTCKLVGINMSKKKSYINRTGTF EFTSFFYRYGFVANFSMELPSFGVSGINESADMSIGVTVIKNNMINNDLGPATAQMALQLFIKDYRYTYR CHRGDTQIQTRRSFELKKLWEQTRSKAGLLVSDGGSNLYNIRNLHIPEVCLKWELMDEDYQGRLCNPLNP FVSHKEIESVNNAVVMPAHGPAKSMEYDAVATTHSWTPKRNRSILNTSQRGILEDEQMYQKCCNLFEKFF PSSSYRRPVGISSMVEAMVSRARIDARIDFESGRIKKEEFAEIMKICSTIEELRRQK SEQIDNO:31 >gi|73921567|ref|YP_308869.1| non-structuralproteinNS2 [InfluenzaAvirus(A/Korea/426/68(H2N2))] MDSNTVSSFQDILLRMSKMQLGSSSEDLNGMITQFESLKLYRDSLGEAVMRMGDLHSLQNRNGKWREQLG QKFEEIRWLIEEVRHRLKITENSFEQITFMQALQLLFEVEQEIRTFSFQLI SEQIDNO:32 >gi|73921566|ref|YP_308870.1| non-structuralproteinNS1 [InfluenzaAvirus(A/Korea/426/68(H2N2))] MDSNTVSSFQVDCFLWHVRKQVVDQELGDAPFLDRLRRDQKSLRGRGSTLDLDIEAATRVGKQIVERILK EESDEALKMTMASAPASRYLTDMTIEELSRDWFMLMPKQKVEGPLCIRIDQAIMDKNIMLKANFSVIFDR LETLILLRAFTEEGAIVGEISPLPSLPGHTIEDVKNAIGVLIGGLEWNDNTVRVSKTLQRFAWRSSNENG RPPLTPKQKRKMARTIRSKVRRDKMAD SEQIDNO:33 >gi|73921307|ref|YP_308871.1| nucleoprotein[InfluenzaAvirus (A/Korea/426/68(H2N2))] MASQGTKRSYEQMETDGERQNATEIRASVGKMIDGIGRFYIQMCTELKLSDYEGRLIQNSLTIERMVLSA FDERRNKYLEEHPSAGKDPKKTGGPIYKRVDGKWMRELVLYDKEEIRRIWRQANNGDDATAGLTHMMIWH SNLNDTTYQRTRALVRTGMDPRMCSLMQGSTLPRRSGAAGAAVKGVGTMVMELIRMIKRGINDRNFWRGE NGRKTRSAYERMCNILKGKFQTAAQRAMMDQVRESRNPGNAEIEDLIFLARSALILRGSVAHKSCLPACV YGPAIASGYNFEKEGYSLVGIDPFKLLQNSQVYSLIRPNENPAHKSQLVWMACNSAAFEDLRVLSFIRGT KVSPRGKLSTRGVQIASNENMDTMESSTLELRSRYWAIRTRSGGNTNQQRASAGQISVQPAFSVQRNLPF DKPTIMAAFTGNTEGRTSDMRAEIIRMMEGAKPEEMSFQGRGVFELSDEKATNPIVPSFDMSNEGSYFFG DNAEEYDN SEQIDNO:34 >gi|73921304|ref|YP_308872.1| neuraminidase[InfluenzaAvirus (A/Korea/426/68(H2N2))] MNPNQKIITIGSVSLTIATVCFLMQIAILVTTVTLHFKQHECDSPASNQVMPCEPIIIERNITEIVYLNN TTIEKEICPEVVEYRNWSKPQCQITGFAPFSKDNSIRLSAGGDIWVTREPYVSCDPGKCYQFALGQGTTL DNKHSNDTIHDRIPHRTLLMNELGVPFHLGTRQVCVAWSSSSCHDGKAWLHVCVTGDDKNATASFIYDGR LMDSIGSWSQNILRTQESECVCINGTCTVVMTDGSASGRADTRILFIEEGKIVHISPLSGSAQHVEECSC YPRYPDVRCICRDNWKGSNRPVIDINMEDYSIDSSYVCSGLVGDTPRNDDRSSNSNCRNPNNERGNPGVK GWAFDNGDDVWMGRTISKDLRSGYETFKVIGGWSTPNSKSQINRQVIVDSNNWSGYSGIFSVEGKRCINR CFYVELIRGRQQETRVWWTSNSIVVFCGTSGTYGTGSWPDGANINFMPI SEQIDNO:35 >gi|73919213|ref|YP_308844.1| nonstructuralprotein2 [InfluenzaAvirus(A/NewYork/392/2004(H3N2))] MDSNTVSSFQDILLRMSKMQLGSSSEDLNGMITQFESLKIYRDSLGEAVMRMGDLHLLQNRNGKWREQLG QKFEEIRWLIEEVRHRLKTTENSFEQITFMQALQLLFEVEQEIRTFSFQLI SEQIDNO:36 >gi|73919212|ref|YP_308845.1| nonstructuralprotein1 [InfluenzaAvirus(A/NewYork/392/2004(H3N2))] MDSNTVSSFQVDCFLWHIRKQVVDQELSDAPFLDRLRRDQRSLRGRGNTLGLDIKAATHVGKQIVEKILK EESDEALKMTMVSTPASRYITDMTIEELSRNWFMLMPKQKVEGPLCIRMDQAIMEKNIMLKANFSVIFDR LETIVLLRAFTEEGAIVGEISPLPSFPGHTIEDVKNAIGVLIGGLEWNDNTVRVSKNLQRFAWRSSNENG GPPLTPKQKRKMARTARSKV SEQIDNO:37 >gi|73919207|ref|YP_308839.1| hemagglutinin[InfluenzaA virus(A/NewYork/392/2004(H3N2))] MKTIIALSYILCLVFAQKLPGNDNSTATLCLGHHAVPNGTIVKTITNDQIEVTNATELVQSSSTGGICDS PHQILDGENCTLIDALLGDPQCDGFQNKKWDLFVERSKAYSNCYPYDVPDYASLRSLVASSGTLEFNNES FNWTGVTQNGTSSACKRRSNNSFFSRLNWLTHLKFKYPALNVTMPNNEKFDKLYIWGVHHPGTDNDQISL YAQASGRITVSTKRSQQTVIPSIGSRPRIRDVPSRISIYWTIVKPGDILLINSTGNLIAPRGYFKIRSGK SSIMRSDAPIGKCNSECITPNGSIPNDKPFQNVNRITYGACPRYVKQNTLKLATGMRNVPEKQTRGIFGA IAGFIENGWEGMVDGWYGFRHQNSEGTGQAADLKSTQAAINQINGKLNRLIGKTNEKFHQIEKEFSEVEG RIQDLEKYVEDTKIDLWSYNAELLVALENQHTIDLTDSEMNKLFERTKKQLRENAEDMGNGCFKIYHKCD NACIGSIRNGTYDHDVYRDEALNNRFQIKGVELKSGYKDWILWISFAISCFLLCVALLGFIMWACQKGNI RCNICI SEQIDNO:38 >gi|73919153|ref|YP_308840.1| matrixprotein2[InfluenzaA virus(A/NewYork/392/2004(H3N2))] MSLLTEVETPIRNEWGCRCNDSSDPLVVAASIIGILHLILWILDRLFFKCVYRLFKHGLKRGPSTEGVPE SMREEYRKEQQNAVDADDSHFVSIELE SEQIDNO:39 >gi|73919152|ref|YP_308841.1| matrixprotein1[InfluenzaA virus(A/NewYork/392/2004(H3N2))] MSLLTEVETYVLSIVPSGPLKAEIAQRLEDVFAGKNTDLEALMEWLKTRPILSPLTKGILGFVFTLTVPS ERGLQRRRFVQNALNGNGDPNNMDKAVKLYRKLKREITFHGAKEIALSYSAGALASCMGLIYNRMGAVTT EVAFGLVCATCEQIADSQHRSHRQMVATTNPLIKHENRMVLASTTAKAMEQMAGSSEQAAEAMEIASQAR QMVQAMRAVGTHPSSSTGLRDDLLENLQTYQKRMGVQMQRFK SEQIDNO:40 >gi|73919150|ref|YP_308848.1| PB1-F2protein[InfluenzaA virus(A/NewYork/392/2004(H3N2))] MEQEQDTPWTQSTEHTNIQRRGSGRQIQKLGHPNSTQLMDHYLRIMSQVDMHKQTVSWRLWPSLKNPTQV SLRTHALKQWKSFNKQGWTN SEQIDNO:41 >gi|73919149|ref|YP_308847.1| polymerasePB1[InfluenzaA virus(A/NewYork/392/2004(H3N2))] MDVNPTLLFLKVPAQNAISTTFPYTGDPPYSHGTGTGYTMDTVNRTHQYSEKGKWTTNTETGAPQLNPID GPLPEDNEPSGYAQTDCVLEAMAFLEESHPGIFENSCLETMEVVQQTRVDKLTQGRQTYDWTLNRNQPAA TALANTIEVFRSNGLTANESGRLIDFLKDVMESMDKEEMEITTHFQRKRRVRDNMTKKMVTQRTIGKKKQ RVNKRGYLIRALTLNTMTKDAERGKLKRRAIATPGMQIRGFVYFVETLARSICEKLEQSGLPVGGNEKKA KLANVVRKMMTNSQDTELSFTITGDNTKWNENQNPRMFLAMITYITKNQPEWFRNILSIAPIMFSNKMAR LGKGYMFESKRMKLRTQIPAEMLASIDLKYFNESTRKKIEKIRPLLIDGTASLSPGMMMGMFNMLSTVLG VSVLNLGQKKYTKTTYWWDGLQSSDDFALIVNAPNHEGIQAGVDRFYRTCKLVGINMSKKKSYINKTGTF EFTSFFYRYGFVANFSMELPSFGVSGINESADMSIGVTVIKNNMINNDLGPATAQMALQLFIKDYRYTYR CHRGDTQIQTRRSFELKKLWDQTQSRAGLLVSDGGPNLYNIRNLHIPEVCLKWELMDENYRGRLCNPLNP FVSHKEIESVNNAVVMPAHGPAKSMEYDAVATTHSWNPKRNRSILNTSQRGILEDEQMYQKCCNLFEKFF PSSSYRRPIGISSMVEAMVSRARIDARIDFESGRIKKEEFSEIMKICSTIEELRRQK SEQIDNO:42 >gi|73919147|ref|YP_308843.1| nucleocapsidprotein [InfluenzaAvirus(A/NewYork/392/2004(H3N2))] MASQGTKRSYEQMETDGDRQNATEIRASVGKMIDGIGRFYIQMCTELKLSDHEGRLIQNSLTIEKMVLSA FDERRNKYLEEHPSAGKDPKKTGGPIYRRVDGKWMRELVLYDKEEIRRIWRQANNGEDATAGLTHIMIWH SNLNDATYQRTRALVRTGMDPRMCSLMQGSTLPRRSGAAGAAVKGIGTMVMELIRMVKRGINDRNFWRGE NGRKTRSAYERMCNILKGKFQTAAQRAMVDQVRESRNPGNAEIEDLIFLARSALILRGSVAHKSCLPACA YGPAVSSGYDFEKEGYSLVGIDPFKLLQNSQIYSLIRPNENPAHKSQLVWMACHSAAFEDLRLLSFIRGT KVSPRGKLSTRGVQIASNENMDNMGSSTLELRSGYWAIRTRSGGNTNQQRASAGQTSVQPTFSVQRNLPF EKSTIMAAFTGNTEGRTSDMRAEIIRMMEGAKPEEVSFRGRGVFELSDEKATNPIVPSFDMSNEGSYFFG DNAEEYDN SEQIDNO:43 >gi|73919136|ref|YP_308842.1| neuraminidase [InfluenzaAvirus(A/NewYork/392/2004(H3N2))] MNPNQKIITIGSVSLTISTICFFMQIAILITTVTLHFKQYEFNSPPNNQVMLCEPTIIERNITEIVYLTN TTIEKEMCPKLAEYRNWSKPQCDITGFAPFSKDNSIRLSAGGDIWVTREPYVSCDPDKCYQFALGQGTTL NNVHSNDTVHDRTPYRTLLMNELGVPFHLGTKQVCIAWSSSSCHDGKAWLHVCVTGDDKNATASFIYNGR LVDSIVSWSKKILRTQESECVCINGTCTVVMTDGSASGKADTKILFIEEGKIIHTSTLSGSAQHVEECSC YPRYPGVRCVCRDNWKGSNRPIVDINIKDYSIVSSYVCSGLVGDTPRKNDSSSSSHCLDPNNEEGGHGVK GWAFDDGNDVWMGRTISEKLRSGYETFKVIEGWSKPNSKLQINRQVIVDRGNRSGYSGIFSVEGKSCINR CFYVELIRGRKEETEVLWTSNSIVVFCGTSGTYGTGSWPDGADINLMPI SEQIDNO:44 >gi|73919134|ref|YP_308846.1| polymerasePA [InfluenzaAvirus(A/NewYork/392/2004(H3N2))] MEDFVRQCFNPMIVELAEKAMKEYGEDLKIETNKFAAICTHLEVCFMYSDFHFINEQGESIVVELDDPNA LLKHRFEIIEGRDRTMAWTVVNSICNTTGAEKPKFLPDLYDYKENRFIEIGVTRREVHIYYLEKANKIKS ENTHIHIFSFTGEEIATKADYTLDEESRARIKTRLFTIRQEMANRGLWDSFRQSERGEETIEEKFEISGT MRRLADQSLPPKFSCLENFRAYVDGFEPNGCIEGKLSQMSKEVNAKIEPFLKTTPRPIKLPNGPPCYQRS KFLLMDALKLSIEDPSHEGEGIPLYDAIKCIKTFFGWKEPYIVKPHEKGINSNYLLSWKQVLSELQDIEN EEKIPRTKNMKKTSQLKWALGENMAPEKVDFDNCRDISDLKQYDSDEPELRSLSSWIQNEFNKACELTDS IWIELDEIGEDVAPIEYIASMRRNYFTAEVSHCRATEYIMKGVYINTALLNASCAAMDDFQLIPMISKCR TKEGRRKTNLYGFIIKGRSHLRNDTDVVNFVSMEFSLTDPRLEPHKWEKYCVLEIGDMLLRSAIGQISRP MFLYVRTNGTSKVKMKWGMEMRRCLLQSLQQIESMIEAESSIKEKDMTKEFFENKSEAWPIGESPKGVEE GSIGKVCRTLLAKSVFNSLYASPQLEGFSAESRKLLLVVQALRDNLEPGTFDLGGLYEAIEECLINDPWV LLNASWFNSFLTHALK SEQIDNO:45 >gi|73919060|ref|YP_308849.1| polymerasePB2 [InfluenzaAvirus(A/NewYork/392/2004(H3N2))] MERIKELRNLMSQSRTREILTKTTVDHMAIIKKYTSGRQEKNPSLRMKWMMAMKYPITADKRITEMVPER NEQGQTLWSKMSDAGSDRVMVSPLAVTWWNRNGPVASTVHYPKVYKTYFDKVERLKHGTFGPVHFRNQVK IRRRVDINPGHADLSAKEAQDVIMEVVFPNEVGARILTSESQLTITKEKKEELRDCKISPLMVAYMLERE LVRKTRFLPVAGGTSSIYIEVLHLTQGTCWEQMYTPGGEVRNDDVDQSLIIAARNIVRRAAVSADPLASL LEMCHSTQIGGTRMVDILRQNPTEEQAVDICKAAMGLRISSSFSFGGFTFKRTSGSSVKKEEEVLTGNLQ TLKIRVHEGYEEFTMVGKRATAILRKATRRLVQLIVSGRDEQSIAEAIIVAMVFSQEDCMIKAVRGDLNF VNRANQRLNPMHQLLRHFQKDAKVLFQNWGIEHIDSVMGMVGVLPDMTPSTEMSMRGIRVSKMGVDEYSS TERVVVSIDRFLRVRDQRGNVLLSPEEVSETQGTERLTITYSSSMMWEINGPESVLVNTYQWIIRNWEAV KIQWSQNPAMLYNKMEFEPFQSLVPKAIRSQYSGFVRTLFQQMRDVLGTFDTTQIIKLLPFAAAPPKQSR MQFSSLTVNVRGSGMRILVRGNSPVFNYNKTTKRLTILGKDAGTLIEDPDESTSGVESAVLRGFLIIGKE DRRYGPALSINELSNLAKGEKANVLIGQGDVVLVMKRKRDSSILTDSQTATKRIRMAIN SEQIDNO:46:CMVProteinIE122: >gi|39841910|gb|AAR31478.1| UL122[Humanherpesvirus5] MESSAKRKMDPDNPDEGPSSKVPRPETPVTKATTFLQTMLRKEVNSQLSLGDPLFPELAEESLKTFEQVT EDCNENPEKDVLAELGDILAQAVNHAGIDSSSTGHTLTTHSCSVSSAPLNKPTPTSVAVTNTPLPGASAT PELSPRKKPRKTTRPFKVIIKPPVPPAPIMLPLIKQEDIKPEPDFTIQYRNKIIDTAGCIVISDSEEEQG EEVETRGATASSPSTGSGTPRVTSPTHPLSQMNHPPLPDPLARPDEDSSSSSSSSCSSASDSESESEEMK CSSGGGASVTSSHHGRGGFGSAASSSLLSCGHQSSGGASTGPRKKKSKRISELDNEKVRNIMKDKNTPFCTPNVQTRRG RVKIDEVSRMFRNTNRSLEYKNLPFTIPSMHQVLDEAIKACKTMQVNNKGIQIIYTRNHEVKSEVDAVRCRLGTMCNLA LSTPFLMEHTMPVTHPPEVAQRTADACNEGVKAAWSLKELHTHQLCPRSSDYRNMIIHAATPVDLLGALNLCLPLMQKF PKQVMVRIFSTNQGGFMLPIYETAAKAYAVGQFEQPTETPPEDLDTLSLAIEAAIQDLRNKSQ SEQIDNO:126: >gi|4927721|gb|AAD33253.1|AF125673_2E7 [Humanpapillomavirustype16] MHGDTPTLHEYMLDLQPETTDLYCYEQLNDSSEEEDEIDGPAGQAEPDRAHYNIVTFCCKCDSTLRLCVQ STHVDIRTLEDLLMGTLGIVCPICSQKP SEQIDNO:200:InfluensaM2 >gi|21693176|gb|AAM75162| /Human/M2/H1N1/Puerto Rico/1934///matrixproteinM2[InfluenzaAvirus (A/PuertoRico/8/34/MountSinai(H1N1))] MSLLTEVETPIRNEWGCRCNGSSDPLAIAANIIGILHLTLWILDRLFFKCIYRRFKYGLKGGPSTEGVPK SMREEYRKEQQSAVDADDGHFVSIELE SEQIDNO:201:>gi|1906383|gb|AAB50256.1| tatprotein [Humanimmunodeficiencyvirus1] MEPVDPRLEPWKHPGSQPKTACTNCYCKKCCFHCQVCFITKALGISYGRKKRRQRRRAHQNSQTHQASLS KQPTSQPRGDPTGPKE SEQIDNO:202:>B.FR.1983.HXB2-LAI-IIIB-BRU(gp120) MRVKEKYQHLWRWGWRWGTMLLGMLMICSATEKLWVTVYYGVPVWKEATTTLFCASDAKAYDTEVHNVWATHACV PTDPNPQEVVLVNVTENFNMWKNDMVEQMHEDIISLWDQSLKPCVKLTPLCVSLKCTDLKNDTNTNSSSGRMIME KGEIKNCSFNISTSIRGKVQKEYAFFYKLDIIPIDNDTTSYKLTSCNTSVITQACPKVSFEPIPIHYCAPAGFAI LKCNNKTFNGTGPCTNVSTVQCTHGIRPVVSTQLLLNGSLAEEEVVIRSVNFTDNAKTIIVQLNTSVEINCTRPN NNTRKRIRIQRGPGRAFVTIGKIGNMRQAHCNISRAKWNNTLKQIASKLREQFGNNKTIIFKQSSGGDPEIVTHS FNCGGEFFYCNSTQLFNSTWFNSTWSTEGSNNTEGSDTITLPCRIKQIINMWQKVGKAMYAPPISGQIRCSSNIT GLLLTRDGGNSNNESEIFRPGGGDMRDNWRSELYKYKVVKIEPLGVAPTKAKRRVVQREKR SEQIDNO:203:HIVgp41 >B.FR.1983.HXB2-LAI-IIIB-BRU(ACCNo.K03455) AVGIGALFLGFLGAAGSTMGAASMTLTVQARQLLSGIVQQQNNLLRAIEAQQHLLQLTVWGIKQLQARILAVERY LKDQQLLGIWGCSGKLICTTAVPWNASWSNKSLEQIWNHTTWMEWDREINNYTSLIHSLIEESQNQQEKNEQELL ELDKWASLWNWFNITNWLWYIKLFIMIVGGLVGLRIVFAVLSIVNRVRQGYSPLSFQTHLPTPRGPDRPEGIEEE GGERDRDRSIRLVNGSLALIWDDLRSLCLFSYHRLRDLLLIVTRIVELLGRRGWEALKYWWNLLQYWSQELKNSA VSLLNATAIAVAEGTDRVIEVVQGACRAIRHIPRRIRQGLERILL SEQIDNO:204:>1b._._.AB016785._(HCV-E1) YEVRNVSGVYHVTNDCSNSSIVYGAADMIMHTPGCVPCVRENNSSRCWVALTPTLAARNRSIPTTTIRRHVDLLV GAAAFCSAMYVGDLCGSVFLVSQLFTFSPRRYETVQDCNCSLYPGHVSGHRMAWDMMMNWSPTAALVVSQLLRIP QAVVDMVTGAHWGVLAGLAYYSMVGNWAKVLIVMLLFAGVDG SEQIDNO:205:>1b._._.AB016785.AB016785 TTHVTGGQTGRTTLGITAMFAFGPHQKLQLINTNGSWHINRTALNCNDSLNTGFLAALFYARKFNSSGCPERMAS CRPIDKFVQGWGPITHAVPDNLDQRPYCWHYAPQPCGIIPASQVCGPVYCFTPSPVVVGTTDRFGAPTYTWGENE TDVLLLNNTRPPQGNWFGCTWMNGTGFAKTCGGPPCNIGGVGNNTLTCPTDCFRKHPEATYTKCGSGPWLTPRCM VDYPYRLWHYPCTVNFTIFKVRMYVGGVEHRLTAACNWTRGERCDLEDRDRSELSPLLLSTTEWQVLPCSFTTLP ALSTGLIHLHQNIVDVQYLYGVGSAVVSIVIKWEYILLLFLLLADARVCACLWMMLLIAQAEA SEQIDNO:309:>gi|52139259|ref|YP_081534.1| major capsidprotein[Humanherpesvirus5] MENWSALELLPKVGIPTDFLTHVKTSAGEEMFEALRIYYGDDPERYNIHFEAIFGTFCNRLEWVYFLTSG LAAAAHAIKFHDLNKLTTGKMLFHVQVPRVASGAGLPTSRQTTIMVTKYSEKSPITIPFELSAACLTYLR ETFEGTILDKILNVEAMHTVLRALKNTADAMERGLIHSFLQTLLRKAPPYFVVQTLVENATLARQALNRI QRSNILQSFKAKMLATLFLLNRTRDRDYVLKFLTRLAEAATDSILDNPTTYTTSSGAKISGVMVSTANVM QIIMSLLSSHITKETVSAPATYGNFVLSPENAVTAISYHSILADFNSYKAHLTSGQPHLPNDSLSQAGAH SLTPLSMDVIRLGEKTVIMENLRRVYKNTDTKDPLERNVDLTFFFPVGLYLPEDRGYTTVESKVKLNDTV RNALPTTAYLLNRDRAVQKIDFVDALKTLCHPVLHEPAPCLQTFTERGPPSEPAMQRLLECRFQQEPMGG AARRIPHFYRVRREVPRTVNEMKQDFVVTDFYKVGNITLYTELHPFFDFTHCQENSETVALCTPRIVIGN LPDGLAPGPFHELRTWEIMEHMRLRPPPDYEETLRLFKTTVTSPNYPELCYLVDVLVHGNVDAFLLIRTF VARCIVNMFHTRQLLVFAHSYALVTLIAEHLADGALPPQLLFHYRNLVAVLRLVTRISALPGLNNGQLAE EPLSAYVNALHDHRLWPPFVTHLPRNMEGVQVVADRQPLNPANIEARHHGVSDVPRLGAMDADEPLFVDD YRATDDEWTLQKVFYLCLMPAMTNNRACGLGLNLKTLLVDLFYRPAFLLMPAATAVSTSGTTSKESTSGV TPEDSIAAQRQAVGEMLTELVEDVATDAHTPLLQACRELFLAVQFVGEHVKVLEVRAPLDHAQRQGLPDF ISRQHVLYNGCCVVTAPKTLIEYSLPVPFHRFYSNPTICAALSDDIKRYVTEFPHYHRHDGGFPLPTAFA HEYHNWLRSPFSRYSATCPNVLHSVMTLAAMLYKISPVSLVLQTKAHIHPGFALTAVRTDTFEVDMLLYS GKSCTSVIINNPIVTKEERDISTTYHVTQNINTVDMGLGYTSNTCVAYVNRVRTDMGVRVQDLFRVFPMN VYRHDEVDRWIRHAAGVERPQLLDTETISMLTFGSMSERNAAATVHGQKAACELILTPVTMDVNYFKIPN NPRGRASCMLAVDPYDTEAATKAIYDHREADAQTFAATHNPWASQAGCLSDVLYNTRHRERLGYNSKFYS PCAQYFNTEEIIAANKTLFKTIDEYLLRAKDCIRGDTDTQYVCVEGTEQLIENPCRLTQEALPILSTTTL ALMETKLKGGAGAFATSETHFGNYVVGEIIPLQQSMLFNS SEQIDNO:310:>gi|52139266|ref|YP_081541.1| tegumentproteinUL16[Humanherpesvirus5] MAWRSGLCETDSRTLKQFLQEECMWKLVGKSRKHREYRAVACRSTIFSPEDDGSCILCQLLLFYRDGEWI LCLCCNGRYQGHYGVGHVHRRRRRICHLPTLYQLSFGGPLGPASIDFLPSFSQVTSSMTCDGITPDVIYE VCMLVPQDEAKRILVKGHGAMDLTCQKAVTLGGAGAWLLPRPEGYTLFFYILCYDLFTSCGNRCDIPSMT RLMAAATACGQAGCSFCTDHEGHVDPTGNYVGCTPDMGRCLCYVPCGPMTQSLIHNEEPATFFCESDDAK YLCAVGSKTAAQVTLGDGLDYHIGVKDSEGRWLPVKTDVWDLVKVEEPVSRMIVCSCPVLKNLVH SEQIDNO:311:>gi|52139212|ref|YP_081485.1| tegumentproteinUL26[Humanherpesvirus5] MTSRRAPDGGLNLDDFMRRQRGRHLDLPYPRGYTLFVCDVEETILTPRDVEYWKLLVVTQGQLRVIGTIG LANLFSWDRSVAGVAADGSVLCYEISRENFVVRAADSLPQLLERGLLHSYFEDVERAAQGRLRHGNRSGL RRDADGQVIRESACYVSRALLRHRVTPGKQEITDAMFEAGNVPSALLP SEQIDNO:312:>gi|52139244|ref|YP_081517.1| multifunctionalexpressionregulator[Humanherpesvirus5] MELHSRGRHDAPSLSSLSERERRARRARRFCLDYEPVPRKFRRERSPTSPSTRNGAAASEYHLAEDTVGA ASHHHRPCVPARRPRYSKDDDTEGDPDHYPPPLPPSSRHALGGTGGHIIMGTAGFRGGHRASSSFKRRVA ASASVPLNPHYGKSYDNDDGEPHHHGGDSTHLRRRVPSCPTTFGSSHPSSANNHHGSSAGPQQQQMLALI DDELDAMDEDELQQLSRLIEKKKRARLQRGAASSGTSPSSTSPVYDLQRYTAESLRLAPYPADLKVPTAF PQDHQPRGRILLSHDELMHTDYLLHIRQQFDWLEEPLLRKLVVEKIFAVYNAPNLHTLLAIIDETLSYMK YHHLHGLPVNPHDPYLETVGGMRQLLFNKLNNLDLGCILDHQDGWGDHCSTLKRLVKKPGQMSAWLRDDV CDLQKRPPETFSQPMHRAMAYVCSFSRVAVSLRRRALQVTGTPQFFDQFDTNNAMGTYRCGAVSDLILGA LQCHECQNEMCELRIQRALAPYRFMIAYCPFDEQSLLDLTVFAGTTTTTASNHATAGGQQRGGDQIHPTD EQCASMESRTDPATLTAYDKKDREGSHRHPSPMIAAAAPPAQPPSQPQQHYSEGELEEDEDSDDASSQDL VRATDRHGDTVVYKTTAVPPSPPAPLAGVRSHRGELNLMTPSPSHGGSPPQVPHKQPIIPVQSANGNHST TATQQQQPPPPPPVPQEDDSVVMRCQTPDYEDMLCYSDDMDD

Example 1

Preparation of Dimeric Peptides According to the Invention

(253) Amino acids that link two monomeric peptide sequences are underlined.

(254) Influenza (M2e):

(255) Constructs derived from the extracellular domain on influenza protein M2 (M2e-domain)

(256) Native Domain:

(257) MSLLTEVETPIRNEWGCRCNDSSD

(258) The following sequences was prepared or are under preparation. The different parts, Z.sup.1-Z.sup.7, are divided by brackets.

(259) ##STR00003##

(260) This construct links the monomeric peptides via a Dpr(Aoa) in the first peptide to an oxidized by NaIO.sub.4 Dpr(Ser) residue in the second.

(261) Dpr(Aoa)=N--Fmoc-N--(N-t.-Boc-amino-oxyacetyl)-L-diaminopropionic acid

(262) Explanation:

(263) The brackets used in the sequences are meant to indicate the different parts/boxes. For the BI155 monomeric parts, the boxes will have the following amino-acid sequences (A/B monomer):

(264) Part Z.sup.1 RG

(265) Part Z.sup.2 Dpr(Aoa)/Dpr(Ser)

(266) Part Z.sup.3 TPI(Har)QDWGNRAN/TPT(Har)NGWDVKLS

(267) Part Z.sup.4 RG

(268) Part Z.sup.5-, means not present in these peptides

(269) Part Z.sup.6 TPTRQEWDCRIS/TPI(Har)QEW(Har)SL(Nle)NQEW

(270) Part Z.sup.7 not present (optional)

(271) The boxes on part of the other sequences can be found in a similar manner

(272) ##STR00004##

(273) Examples of disulfide linked constructs can be, but are not restricted to, the following linked peptide sequences:

(274) ##STR00005##

(275) The above disulfide linked constructs may e.g. be synthesised by titration of 2-pyridinesulfenyl (SPyr)-protected cysteine-containing peptides with thiol-unprotected peptides. This has proven to be a superior procedure to selectively generate disulfide-linked peptide heterodimers preventing the formation of homodimers (Schutz A et al., Tetrahedron, Volume 56, Issue 24, 9 Jun. 2000, Pages 3889-3891). Similar dimeric constructs may be made with the other monomeric peptides according to the invention.

(276) ##STR00006##

(277) Examples of thio-esther linked constructs can be, but are not restricted to, the following linked peptide sequences:

(278) ##STR00007##

(279) The Cys-Lys linker is typically established in the form of a thioether bond between a cysteine in one peptide and a bromoacetyl derivatized lysine in the other peptide.

(280) Examples of other linked constructs can be, but are not restricted to, the following linked peptide sequences, N--methylated Lys may be linked to Asp or Glu by a side-chain to side-chain peptide bond, wherein the N methylation makes the bond more stable (Lys(Me) refers to an N--methylated Lys residue).

(281) ##STR00008##

Example 3

Immunological Studies

(282) Rabbit Immunizations

(283) New Zealand White female rabbits (n=3) is immunized intradermally at weeks 0, 2 & 6 with 1 ml of BI400-B vaccine consisting of 500 g BI400-B in 50% V/V Freund's adjuvant (i.e. Complete Freund's adjuvant used for priming, followed by boostings with Incomplete Freund's adjuvant). Individual blood serum is isolated for ELISA.

(284) Direct ELISA for Human or Rabbit Sera

(285) 50-100 l of BI400-B (pre-incubated in Coating buffer0.05M Na.sub.2CO.sub.3 pH9.6; denoted CBin cold at 16 g/ml for each peptide 1-3 days prior to coating) or just CB (background control) is used for coating wells in microtiter plates at 4 C. overnight. The microtiter plates are then washed 3 with washing buffer (PBS+1% v/v Triton-X100; denoted WB), followed by 2 h blocking at room temperature (RT) with 200 l/well of blocking buffer (PBS+1% w/v BSA). Plates are then washed 3 with WB, followed by 1 h incubation at 37 C. with 50-70 ul/well of added human (or rabbit) sera (serial dilutions ranging from 1:1-1:250 in dilution buffer (PBS+1% v/v Triton-X100+1% w/v BSA; denoted DB)). Plates are then washed 6 with WB, followed by 1 h incubation at RT with 70 l/well of Alkaline Phosphatase-conjugated Protein G (3 g/ml in DB; Calbiochem 539305). Plates are then washed 6 with WB, followed by 10-60 min incubation at room temperature with 100 l/well of 0.3% w/v of Phenophtalein monophosphate (Sigma P-5758). Plates are finally quenched by adding 100 l/well of Quench solution (0.1M TRIS+0.1M EDTA+0.5M NaOH+0.01% w/v NaN.sub.3; pH14), followed by ELISA reader (ASYS UVM 340) at 550 nm.

Example 4

Virus Specific Response by ELISPOT Assay

(286) At day one, PBMC samples from blood donors are thawed, washed with warm medium and incubated in flasks (250000 PBMCs/cm2) for 24 hours at 37 C., 5% CO2 in covering amount of culture media (RPMI 1640 with ultra-glutamine, Lonza, BE12-702F701; 10% Foetal Bovine serum (FBS), Fisher Scientific Cat. No. A15-101; Penicillin/Streptomycin, Fisher Scientific Cat. No. P11-010) to allow the cells to recover after thawing. At day two, the cells are added to a Falcon Microtest Tissue Culture plate, 96 well flat bottom, at 500 000 cells per well in a volume of 200 l total medium. Parallel wells are added the indicated stimuli in duplicate or left with medium as a control for 6 days at 37 C., 5% CO.sub.2. After the six day of incubation, 100 l of the cell suspension are transferred to an ELISPOT (Millipore multiscreen HTS) plate coated with 1 g/ml native influenza M2e protein. After a 24 hour incubation, the plate is washed four times with PBS+0.05% Tween20, and a fifth time with PBS, 200 l/well. A mouse Anti-human IgG or IgM biotin (Southern Biotech 9040-08 and 9020-08) is diluted in PBS with 0.5% FBS and incubated for 90 minutes at 37 C. The washing is repeated as described, before 80 l Streptavidin-Alkaline-Phosphatase (Sigma Aldrich, S289) is added each well and incubated at 60 minutes in the dark, at room temperature. The wells are then washed 2 times with PBS+0.05% Tween20 and 4 times with PBS, 200 l/well, before the substrate, Vector Blue Alkaline Phosphatase Substrate kit III (Vector Blue, SK-5300) is added and let to develop for 7 minutes at room temperature. The reaction is stopped with running water, the plates let dry and the sport enumerated by an ELISPOT reader (CTL-ImmunoSpot S5 UV Analyzer).

(287) Virus Specific Response by ELISA

(288) 100 l of antigen as indicated (pre-incubated in Coating buffer0.05M Na.sub.2CO.sub.3 pH9.6; denoted CBin cold at 8 g/ml 1-3 days) or just CB (background control) is used for coating wells in microtiter plates at 4 C. The microtiter plates are then washed 3 with washing buffer (PBS+1% v/v Triton-X100; denoted WB), followed by 2 h blocking at room temperature (RT) with 200 l/well of blocking buffer (PBS+1% w/v BSA). Plates are then washed 3 with WB, followed by 1 h incubation at 37 C. with 50-70 ul/well of added human (or rabbit or sheep) sera (serial dilutions ranging from 1:5-1:250 in dilution buffer (PBS+1% v/v Triton-X100+1% w/v BSA; denoted DB)). Plates are then washed 6 with WB, followed by 1 h incubation at RT with 70 l/well of Alkaline Phosphatase-conjugated Protein G (3 g/ml in DB; Calbiochem 539305) or goat anti-mouse IgG biotin (1 g/ml, Southern Biotech, 1030-08. In case of the goat anti-mouse IgG biotin, the plates are washed one extra step as described, before addition of 100 l Streptavidin-Alkaline-Phosphatase (1 g/ml, Sigma Aldrich, S289) and incubated 1 hour at RT. Plates are then washed 6 with WB, followed by 10-60 min incubation at room temperature with 100 l/well of 0.3% w/v of Phenophtalein monophosphate (Sigma P-5758). Plates are finally quenched by adding 100 l/well of Quench solution (0.1M TRIS+0.1M EDTA+0.5M NaOH+0.01% w/v NaN.sub.3; pH14), followed by a measurement with a ELISA reader (ASYS UVM 340) at 550 nm. The strength of the sera, i.e. the magnitude of the humoral immune response, is then reported as the dilution of sera that result in the described Optical Density (OD) value, or the OD value at the indicated dilution of sera.

Example 5

(289) The peptides according to the invention used in the following examples are synthesized by Schafer-N as c-terminal amides using the Fmoc-strategy of Sheppard, (1978) J. Chem. Soc., Chem. Commun., 539.

(290) BI100-190e, BI100-190f, BI100-260b, BI100-260c, BI100-260d, BI100-260e, and BI100-260f were synthezised by Schafer-N with and without Biotin in the C-terminal tested:

(291) Cell Penetration Assay

(292) A set of peptides were biotinylated on C-terminal, and different combinations of aminoacids, with respect to length and type, were added to the sequence box Z.sup.1, Z.sup.4 and Z.sup.7 in the peptides according to the present invention, formula I. The peptides were tested on cells grown from one individual blood donor.

(293) Schematic diagram of amino acid sequence of the peptides according to the invention (Each Z here defines a sequence of amino acids):

(294) ##STR00009##
Intracellular Staining for Biotinylated Peptides

(295) 96-well U-bottom polystyrene plates (NUNC, cat no: 163320) were used for staining of human PBMCs. Briefly, 8 ul of N- or C-terminally biotinylated peptides according to table 1 or table 2 (i.e. 5 mM, 2.5 mM & 1.25 mM tested for each peptide) were incubated at 37 C. for 2 h with 40 ul of PBMC (12.5106 cells/ml) from blood donors. Cells were then washed 3 with 150 ul of Cellwash (BD, cat no: 349524), followed by resuspension of each cell pellet with 100 ul of Trypsin-EDTA (Sigma, cat no: T4424), then incubated at 37 C. for 5 min. Trypsinated cells were then washed 3 with 150 ul of Cellwash (BD, cat no: 349524), followed by resuspension with BD Cytofix/Cytoperm plus (BD, cat no: 554715), then incubated at 4 C. for 20 min according to manufacturer. Cells were then washed 2 with 150 ul PermWash (BD, cat no: 554715). Cells were then stained with Streptavidin-APC (BD, cat no: 554067) & Anti-hCD11c (eBioscience, cat no: 12-0116) according to manufacturer at 4 C. for 30 min aiming to visualize biotinylated peptides & dendritic cells, respectively. Cells were then washed 3 with 150 ul PermWash, followed by resuspension in staining buffer (BD, cat no: 554656) before flow cytometry. Dendritic cells were gated as CD11c+ events outside lymphocyte region (i.e. higher FSC & SSC signals than lymphocytes). 200 000 total cells were acquired on a FACSCanto II flow cytometer with HTS loader, and histograms for both total cells & dendritic cells with respect to peptide-fluorescence (i.e. GeoMean) were prepared.

(296) Extracellular Staining for Biotinylated Peptides

(297) 96-well U-bottom polystyrene plates (NUNC, cat no: 163320) were used for staining of human PBMCs. Briefly, 8 ul of N- or C-terminally biotinylated peptides according to table 1 or table 2 (i.e. 5 mM, 2.5 mM & 1.25 mM tested for each peptide; all peptides manufactured by Schafer) were incubated at 37 C. for 2 h with 40 ul of PBMC (12.5106 cells/ml) from blood donors. Cells were then washed 3 with 150 ul of Cellwash (BD, cat no: 349524), then stained with Streptavidin-APC (BD, cat no: 554067) & Anti-hCD11c (eBioscience, cat no: 12-0116) according to manufacturer at 4 C. for 30 min aiming to visualize biotinylated peptides & dendritic cells, respectively. Cells were then washed 3 with 150 ul of Cellwash (BD, cat no: 349524), followed by resuspension in staining buffer (BD, cat no: 554656) before flow cytometry. Dendritic cells were gated as CD11c+ events outside lymphocyte region (i.e. higher FSC & SSC signals than lymphocytes). 200 000 total cells were acquired on a FACSCanto II flow cytometer with HTS loader, and histograms for both total cells & dendritic cells with respect to peptide-fluorescence (i.e. GeoMean) were prepared.

Example 6

(298) Positive CTL response may alternatively be assayed by ELISPOT assay.

Human IFN-Gamma Cytotoxic T-Cell (CTL) Response by ELISPOT Assay

(299) Briefly, at day 1, PBMC samples from HCV patients were incubated in flasks (430 000 PBMCs/cm2) for 2 h at 37 C., 5% CO2 in covering amount of culture media (RPMI 1640 Fisher Scientific; Cat No. PAAE15-039 supplemented with L-Glutamine, (MedProbe Cat. No. 13E17-605E, 10% Foetal Bovine serum (FBS), Fisher Scientific Cat. No. A15-101) and Penicillin/Streptomycin, (Fisher Scientific Cat. No. P11-010) in order to allow adherence of monocytes. Non-adherent cells were isolated, washed, and frozen in 10% V/V DMSO in FBS until further usage. Adherent cells were carefully washed with culture media, followed by incubation at 37 C. until day 3 in culture media containing 2 g/ml final concentration of hrGM-CSF (Xiamen amoytop biotech co, cat no: 3004.9090.90) & 1 g/ml hrIL-4 (Invitrogen, Cat no: PHC0043), and this procedure is then repeated at day 6. At day 7, cultured dendritic cells (5 000-10 000 per well) were added to ELISPOT (Millipore multiscreen HTS) plates coated with 0.5 g/well anti-human Interferon together with thawed autologous non-adherent cells (200 000 per well), antigen samples (1-8 ug/ml final concentration for peptide antigens; 5 ug/ml final concentration for Concanavalin A (Sigma, Cat no: C7275) or PHA (Sigma, Cat no: L2769)) and optionally, anti-Anergy antibodies (0.03-0.05 ug/ml final concentration for both anti-PD-1 (eBioscience, cat no: 16-9989-82) & anti-PD-L1 (eBioscience, cat no: 16-5983-82)). Plates were incubated overnight and spots were developed according to manufacturer. Spots were read on ELISPOT reader (CTL-ImmunoSpot S5 UV Analyzer).

Example 7

The REVEAL & ProVE Rapid Epitope Discovery System in Detail

(300) Binding properties to HLA for the ninemers listed are tested for the following HLA-classes: HLA-A1, HLA-A2, HLA-A3, HLA-A11, HLA-A24, HLA-A29, HLA-B7, HLA-B8, HLA-B14, HLA-B15, HLA-B27, HLA-B35, HLA-B40.

(301) The peptides are synthesized as a Prospector PEPscreen: Custom Peptide Library. Peptides 8-15 amino acids in length are synthesized in 0.5-2 mg quantities with high average purity. Quality control by MALDI-TOF Mass Spectrometry is carried out on 100% of samples.

(302) The REVEAL binding assay determined the ability of each candidate peptide to bind to one or more MHC class I alleles and stabilizing the MHC-peptide complex. By comparing the binding to that of high and intermediate affinity T cell epitopes, the most likely immunogenic peptides in a protein sequence can be identified. Detection is based on the presence or absence of the native conformation of the MHC-peptide complex.

(303) Each peptide is given a score relative to the positive control peptide, which is a known T cell epitope. The score of the test peptide is reported quantitatively as a percentage of the signal generated by the positive control peptide, and the peptide is indicated as having a putative pass or fail result. Assay performance is confirmed by including an intermediate control peptide that is known to bind with weaker affinity to the allele under investigation.

Example 8

Intracellular Staining

(304) Peptides as described herein with Z.sup.3 and Z.sup.6 derived from HCV, Influenza, or CMV are prepared and tested for intracellular staining in an experiment as described above in the Cell penetration assay.

(305) Average over results from buffy coats from ten donors, normalized to N-biotin for each donor is illustrated in FIGS. 3 and 4.

Example 9

(306) TABLE-US-00023 TABLE10 Peptidesusedascontrolsandnotparttotheinvention, butcarryingthesameepitopes(Z3,Z6,Z9)linkedby glycinesandserines,forcomparisontopeptidesofthe invention.(Z= Norleucine,X= Homoarginine,biotc indicatesthatabiotinylatedlysineresiduehasbeen addedtotheC-terminal). Peptide - Z3 - Z6 - Z9 C-tertag BI330-72-2- GS VITYSIFLIVS GS GGNVIGGIYZIPR biotin- ns-biotc NH2 BI330-83-ns- GS TANWARVIS GS ANWAKVIL S NWAKVI biotin- biotc NH2 BI310-511-ns- S GYLPAVGAPI GS VIRVIAHGLRL biotin- biotc NH2 BI100-330-ns- GS TAYERZCNIL GS GLEPLVIAGILA biotin- biotc NH2 BI100-270-ns- GS TVIGASZIPLL GS TPIXQDWENRAN biotin- biotc NH2 BI100-130-ns- GS AAFEEZXITS GS VAFEDLXZZSFI biotin- biotc NH2
Results

(307) Biotinylated versions of scaffold peptides were tested for intracellular and extracellular uptake. All tested peptides had stronger intracellular and extracellular uptake compared to the control peptide N-biotin (N-bio), as seen from FIGS. 1-2. Also when comparing the uptake of peptides according to the invention to peptides carrying the same epitopes linked by Glycine and Serine residues instead (Table 11), tested peptides according to the invention generally had a higher uptake. Many of the peptides tested show very strong uptake and potentially we are seeing saturation of the cell assay system for these.

(308) Values represent averages over readouts from buffy coats from ten (five) donors and three (four) concentrations of peptide each, normalized by value for N-biotin for each donor for scaffold (non-scaffold) peptides respectively.

(309) TABLE-US-00024 TABLE 11 Intracellular and extracellular uptake of peptides of the invention (Bold) compared to peptides containing the same epitopes linked by Gly and Ser residues (non- bold Italics). Median readouts from buffy coats from ten (five) donors and three (four) concentrations of peptide each, normalized by value for N-biotin for each donor for scaffold (non-scaffold) peptides. Peptide Intracellular Extracellular (biotinylated) Uptake Uptake BI100-270 2.05 20.47 BI100-270b 3.35 16.54 BI100-270c 2.91 9.56 BI100-270d 5.73 4.77 BI100-270e 10.26 3.54 BI100-270ns 1.30 1.29 BI100-330 70.36 655.35 BI100-330b 76.42 744.11 BI100-330c 880.85 244.29 BI100-330d 80.82 592.82 BI100-330e 23.89 529.05 BI100-330ns 1.82 416.04 BI310-511 22.62 227.46 BI310-511b 67.29 466.71 BI310-511c 31.83 203.62 BI310-511d 70.64 267.15 BI310-511e 44.59 473.80 BI310-511f 26.85 178.61 BI310-511g 66.74 171.31 BI310-511ns 3.85 4.56 BI330-83 194.69 364.04 BI330-83b 120.10 518.60 BI330-83c 154.43 435.66 BI330-83d 52.14 267.38 B1330-83ns 63.51 380.25

Example 10

Effect of Peptide Based Influenza Vaccine in Protection of HLA A2 Mice Against Influenza Virus Challenge

(310) C57/B6/Tg HLA A2 mice (n=10 mice per group) were immunized week 0 and week 2 by subcutaneous administration (250 l; each side of base of tail), of a solution containing 50 g of each peptide, or 0.07 g HA of inactivated influenza A/PR8 (H1N1) virus given as vaccine control.

(311) At week 4 the mice was infected with live influenza virus in order to measure the immune response to viral infection. The challenge was done with a mouse adapted strain of influenza A at a dose of 110{circumflex over ()}5TCID.sub.50/mouse which is enough to reliably infect the animals without mortality as determined by titration in the same mouse strain. The animals were then monitored for 7 days by weight loss at the start of challenge and daily from day three before they were sacrificed and serum collected. Individual serum for mice in all groups were collected before start of experiment, and day of sacrifice.

(312) TABLE-US-00025 TABLE 12 Group Treatment 1 Vaccinate with peptides + adjuvant Provax (week 0, 2) 2 Vaccinate with peptides + adjuvant ISA 51 (week 0, 2) 3 Vaccinate with inactivated conventional vaccine (week 0, 2) 4 Nave mice.

(313) TABLE-US-00026 TABLE 13 Survival (n) Day Group 1-4 5 6 7 1 Peptide, Provax 10 10 10 7 2 Peptide, ISA 51 10 10 10 9 3 PR8 10 9 9 6 4 Nave 10 10 10 8
Results

(314) Following the weight loss after challenge a clear protective effect is seen for both groups receiving the peptide vaccine with either ISA51 or Provax as adjuvant, as compared to the standard inactivated viral vaccine, PR8, or nave mice (FIG. 5).