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MUTANT OF RSV F PROTEIN

20260117202 ยท 2026-04-30

    Inventors

    Cpc classification

    International classification

    Abstract

    Disclosed is a mutant of a wild-type RSV F protein. The binding experiment of a pre-fusion conformation trimer-specific monoclonal antibody AM14 shows that the mutant provided in embodiments can have strong activity of binding to the AM14 monoclonal antibody, while the wild-type RSV F protein before artificial mutation has no activity of binding to the AM14 monoclonal antibody. The accelerated stability test result shows that the activity of binding of the mutant provided in the embodiments when placed in a 37 C. environment for four weeks to the AM14 monoclonal antibody does not change obviously. The animal immune experiment result shows that compared with the wild-type RSV F protein before artificial mutation, the mutant provided in the embodiments can induce production of a neutralizing antibody having a higher titer.

    Claims

    1-43. (canceled)

    44. A mutant of a wild-type respiratory syncytial virus (RSV) fusion (F) protein, being an artificial mutant, wherein an amino acid sequence of the mutant is a sequence shown in SEQ ID No. 156 or 158-166.

    45. A trimer of the mutant of a wild-type respiratory syncytial virus fusion protein according to claim 44, wherein the trimer is a homologous trimer.

    46. The trimer according to claim 45, wherein a trimerization domain is directly or indirectly linked to the C-terminus of the mutant.

    47. The trimer according to claim 46, wherein the trimerization domain is a GCN4 leucine zipper, collagen or T4 foldon.

    48. The trimer according to claim 46, wherein the C-terminus of the mutant is linked to the trimerization domain via a linker.

    49. The trimer according to claim 48, wherein the linker is a GS repeat sequence, a GGS repeat sequence, an SAIG repeat sequence, a GSS repeat sequence or a GG repeat sequence.

    50. Fusion protein, comprising the mutant according to claim 44, and a signal peptide or tag, wherein the tag is a Flag tag, enhanced green fluorescent protein, flutathione S-transferase, His-tag, streptomycin tag II or AviTag.

    51. Virus-like particles, comprising the mutant according to claim 44.

    52. Virus-like particles, comprising the trimer according to claim 45.

    53. Virus-like particles, comprising the fusion protein according to claim 50.

    54. An isolated nucleic acid molecule, encoding the mutant according to claim 44.

    55. A vector, comprising a sequence of the nucleic acid molecule according to claim 54.

    56. A vector, comprising the mutant according to claim 44.

    57. The vector according to claim 55, being a protein expression vector or a gene delivery vector.

    58. The vector according to claim 56, being a protein delivery vector.

    59. The vector according to claim 57, wherein the gene delivery vector is a viral vector or a non-viral vector; the viral vector comprises a retrovirus vector, a lentivirus vector, an adenovirus vector, an adeno-associated virus vector, a poxvirus vector, a herpes simplex virus vector or a Sendai virus vector; and the non-viral vector comprises a liposome, a cationic polymer vector, a lipid nanoparticle vector or a multifunctional envelope-type nano vector.

    60. A host cell, comprising the nucleic acid molecule according to claim 54.

    61. A host cell, comprising the vector according to claim 55.

    62. A host cell, comprising the vector according to claim 56.

    63. A pharmaceutical composition, comprising the mutant according to claim 44, and a pharmaceutically acceptable carrier.

    64. A pharmaceutical composition, comprising the isolated nucleic acid molecule according to claim 54, and a pharmaceutically acceptable carrier.

    Description

    BRIEF DESCRIPTION OF DRAWINGS

    [0283] FIG. 1A is a structural diagram of a prefusion RSV F protein trimer, and FIG. 1B is a structural diagram of a postfusion RSV F protein trimer, where a head domain and a tail domain defined for visual description in the present disclosure are marked in the figures, refolding region 1 (RR1, located in the head domain) and refolding region 2 (RR2, located in a C-terminal of the head domain and the tail domain) that undergo large conformational changes during a transition of the F protein from a prefusion conformation to a postfusion conformation are marked in the figures, and FIG. 1A and FIG. 1B are drawn based on coordinate files with protein data bank (PDB) identifiers 4jhw and 3rrr respectively;

    [0284] FIG. 2 is a structural superposition diagram of a prefusion conformation (lighter color) and a postfusion conformation (darker color) of an RSV F protein monomer, where during a transition of the F protein from the prefusion conformation to the postfusion conformation, RR1 of a head domain undergoes a large conformational change, and 1 and 5 correspondingly undergo obvious conformational movement; and an enlarged diagram on the right shows a conformational change before and after fusion of 1 and 5, the prefusion conformation and the postfusion conformation of the F protein monomer are drawn according to coordinate files with PDB identifiers 4jhw and 3rrr respectively;

    [0285] FIG. 3 shows sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) diagrams of illustrative mutants FHM-24 and FHM-25 with mutation introduced into head domains according to an example of the present disclosure;

    [0286] FIG. 4 is a graph showing results of AM14 monoclonal antibody binding experiments of illustrative mutants FHM-24 and FHM-25 with mutation introduced into head domains, a wild-type RSV F protein 1 before artificial mutation, and a wild-type RSV F protein 2 before artificial mutation according to an example of the present disclosure;

    [0287] FIG. 5 is a graph showing results of Synagis binding experiments of illustrative mutants FHM-24 and FHM-25 with mutation introduced into head domains, a wild-type RSV F protein 1 before artificial mutation, and a wild-type RSV F protein 2 before artificial mutation according to an example of the present disclosure;

    [0288] FIG. 6 shows SDS-PAGE diagrams of mutants FHCS-1, FHCS-2, FHCS-3, FHCS-4, FHCM-5, FHCM-1, FHCM-2, FHCM-3 and FHCM-4, with purified proteins, constructed in an example of the present disclosure;

    [0289] FIG. 7A is a comparison diagram of results of AM14 monoclonal antibody binding experiments of mutants FHCS-1, FHCS-2, FHCS-3, FHCS-4, FHCS-5, FHCM-1 and FHCM-2 constructed in an example of the present disclosure, a mutant FHM-24, wild-type RSV F protein 1 before artificial mutation, and a wild-type RSV F protein 2 before artificial mutation; and FIG. 7B is a comparison diagram of results of AM14 monoclonal antibody binding experiments of mutants FHCM-3 and FHCM-4 constructed in an example of the present disclosure, a mutant FHM-25, a wild-type RSV F protein 1 before artificial mutation, and a wild-type RSV F protein 2 before artificial mutation;

    [0290] FIG. 8A is a comparison diagram of results of Synagis binding experiments of mutants FHCS-1, FHCS-2, FHCS-3, FHCS-4, FHCS-5, FHCM-1 and FHCM-2 constructed in an example of the present disclosure, a mutant FHM-24, a wild-type RSV F protein 1 before artificial mutation, and a wild-type RSV F protein 2 before artificial mutation; and FIG. 8B is a comparison diagram of results of Synagis binding experiments of mutants FHCM-3 and FHCM-4 constructed in an example of the present disclosure, a mutant FHM-25, a wild-type RSV F protein 1 before artificial mutation, and a wild-type RSV F protein 2 before artificial mutation; and

    [0291] FIG. 9 is a comparison diagram of detection results of live RSV strain A2 neutralizing antibodies of mutants FHCS-1, FHCS-2, FHCS-3 and FHCS-4 constructed in an example of the present disclosure, a mutant FHM-24, and a wild-type RSV F protein 2 before artificial mutation.

    SEQUENCE TABLE

    [0292] Illustrative sequences provided in the present disclosure are shown in Tables 1 to 3.

    TABLE-US-00001 TABLE1 Aminoacidsequenceofwild-typeRSVFprotein Wild-type sequence nameand SEQID description No. Aminoacidsequence Full-length 1 MELLILKANAITTILTAVTFCFASGQNITEEFYQSTCSAVS aminoacid KGYLSALRTGWYTSVITIELSNIKENKCNGTDAKVKLIK sequence QELDKYKNAVTELQLLMQSTPATNNRARRELPRFMNYT ofF LNNAKKTNVTLSKKRKRRFLGFLLGVGSAIASGVAVSK proteinof VLHLEGEVNKIKSALLSTNKAVVSLSNGVSVLTSKVLDL RSVstrain KNYIDKQLLPIVNKQSCSISNIETVIEFQQKNNRLLEITRE A2(Ref: FSVNAGVTTPVSTYMLTNSELLSLINDMPITNDQKKLMS Mclellan NNVQIVRQQSYSIMSIIKEEVLAYVVQLPLYGVIDTPCW J.S.,etal. KLHTSPLCTTNTKEGSNICLTRTDRGWYCDNAGSVSFFP Science, QAETCKVQSNRVFCDTMNSLTLPSEVNLCNVDIFNPKY 2013, DCKIMTSKTDVSSSVITSLGAIVSCYGKTKCTASNKNRGI 342: IKTFSNGCDYVSNKGVDTVSVGNTLYYVNKQEGKSLYV 592-598) KGEPIINFYDPLVFPSDEFDASISQVNEKINQSLAFIRKSD ELLHNVNAGKSTTNIMITTIIIVIIVILLSLIAVGLLLYCKA RSTPVTLSKDQLSGINNIAFSN Full-length 2 MELLILKANAITTILTAVTFCFASGQNITEEFYQSTCSAVS aminoacid KGYLSALRTGWYTSVITIELSNIKENKCNGTDAKVKLIK sequence QELDKYKNAVTELQLLMQSTPPTNNRARRELPRFMNYT ofF LNNAKKTNVTLSKKRKRRFLGFLLGVGSAIASGVAVSK proteinof VLHLEGEVNKIKSALLSTNKAVVSLSNGVSVLTSKVLDL RSVstrain KNYIDKQLLPIVNKQSCSISNIETVIEFQQKNNRLLEITRE A2 FSVNAGVTTPVSTYMLTNSELLSLINDMPITNDQKKLMS (UniProtKB/ NNVQIVRQQSYSIMSIIKEEVLAYVVQLPLYGVIDTPCW Swiss-Prot: KLHTSPLCTTNTKEGSNICLTRTDRGWYCDNAGSVSFFP P03420.1) QAETCKVQSNRVFCDTMNSLTLPSEINLCNVDIFNPKYD CKIMTSKTDVSSSVITSLGAIVSCYGKTKCTASNKNRGII KTFSNGCDYVSNKGMDTVSVGNTLYYVNKQEGKSLYV KGEPIINFYDPLVFPSDEFDASISQVNEKINQSLAFIRKSD ELLHNVNAGKSTTNIMITTIIIVIIVILLSLIAVGLLLYCKA RSTPVTLSKDQLSGINNIAFSN Full-length 3 MELPIIKTNAITAILAAVTLCFASSQNITEEFYQSTCSAVS aminoacid KGYLSALRTGWYTSVITIELSNIKENKCNGTDAKVKLIK sequence QELDKYKNAVTELQLLMQSTPAANSRARRELPRFMNYT ofF LNNTKNTNVTLSKKRKRRFLGFLLGVGSAIASGIAVSKV proteinof LHLEGEVNKIKSALLSTNKAVVSLSNGVSVLTSKVLDLK RSVstrain NYIDKQLLPIVNKQSCSISNIETVIEFQQKNNRLLEITREF ON1of SVNAGVTTPVSTYMLTNSELLSLINDMPITNDQKKLMSS subtypeA NVQIVRQQSYSIMSIIKEEVLAYVVQLPLYGVIDTPCWK (GenBank: LHTSPLCTTNTKEGSNICLTRTDRGWYCDNAGSVSFFPQ QYW11427.1) AETCKVQSNRVFCDTMNSLTLPSEVNLCNIDIFNPKYDC KIMTSKTDVSSSVITSLGAIVSCYGKTKCTASNKNRGIIK TFSNGCDYVSNKGVDTVSVGNTLYYVNKQEGKSLYVK GEPIINFYDPLVFPSDEFDASISQVNEKINQSLAFIRKSDE LLHNVNAGKSTTNIMITTIIIVIIVILLALIAVGLLLYCKAR STPVTLSKDQLSGINNIAFSN Full-length 4 MELLIHRSSAIFLTLAINALHLTSSQNITEEFYQSTCSAVS aminoacid RGYLSALRTGWYTSVITIELSNIKETKCNGTDTKVKLIK sequence QELDKYKNAVTELQLLMQNTPAVNNRARREAPQYMNY ofF TINTTKNLNVSISKKRKRRFLGFLLGVGSAIASGIAVSKV proteinof LHLEGEVNKIKNALQLTNKAVVSLSNGVSVLTSRVLDLK RSVstrain NYINNQLLPMVNRQSCRISNIETVIEFQQKNSRLLEITRE BAof FSVNAGVTTPLSTYMLTNSELLSLINDMPITNDQKKLMS subtypeB SNVQIVRQQSYSIMSIIKEEVLAYVVQLPIYGVIDTPCWK (GenBank: LHTSPLCTTNIKEGSNICLTRTDRGWYCDNAGSVSFFPQ QYW12940.1) ADTCKVQSNRVFCDTMNSLTLPSEVSLCNTDIFNSKYDC KIMTSKTDISSSVITSLGAIVSCYGKTKCTASNKNRGIIKT FSNGCDYVSNKGVDTVSVGNTLYYVNKLEGKNLYVKG EPIINYYDPLVFPSDEFDASISQVNEKINQSLAFIRRSDEL LHNVNTGKSTTNIMITAIIIVIIVVLLSLIAIGLLLYCKAKN TPVTLSKDQLSGINNIAFSK Full-length 5 MELLIHRSSAIFLTLAINALYLTSSQNITEEFYQSTCSAVS aminoacid RGYFSALRTGWYTSVITIELSNIKETKCNGTDTKVKLIK sequence QELDKYKNAVTELQLLMQNTPAANNRARREAPQYMNY ofF TINTTKNLNVSISKKRKRRFLGFLLGVGSAIASGIAVSKV proteinof LHLEGEVNKIKNALLSTNKAVVSLSNGVSVLTSKVLDLK RSVof NYINNQLLPIVNQQSCRISNIETVIEFQQKNSRLLEITREF subtypeB SVNAGVTTPLSTYMLTNSELLSLINDMPITNDQKKLMSS (GenBank: NVQIVRQQSYSIMSIIKEEVLAYVVQLPIYGVIDTPCWKL AHV80758) HTSPLCTTNIKEGSNICLTRTDRGWYCDNAGSVSFFPQA DTCKVQSNRVFCDTMNSLTLPSEVSLCNTDIFNSKYDCK IMTSKTDISSSVITSLGAIVSCYGKTKCTASNKNRGIIKTF SNGCDYVSNKGVDTVSVGNTLYYVNKLEGKNLYVKGE PIINYYDPLVFPSDEFDASISQVNEKINQSLAFIRRSDELL HNVNTGKSTTNIMITAIIIVIIVVLLSLIAIGLLLYCKAKNT PVTLSKDQLSGINNIAFSK Full-length 6 MELLIHRSSAIFLTLAVNALYLTSSQNITEEFYQSTCSAVS aminoacid RGYFSALRTGWYTSVITIELSNIKETKCNGTDTKVKLIK sequence QELDKYKNAVTELQLLMQNTPAANNRARREAPQYMNY ofF TINTTKNLNVSISKKRKRRFLGFLLGVGSAIASGIAVSKV proteinof LHLEGEVNKIKNALLSTNKAVVSLSNGVSVLTSKVLDLK RSVof NYINNRLLPIVNQQSCRISNIETVIEFQQMNSRLLEITREF subtypeB SVNAGVTTPLSTYMLTNSELLSLINDMPITNDQKKLMSS (UniProtKB/ NVQIVRQQSYSIMSIIKEEVLAYVVQLPIYGVIDTPCWKL Swiss-Prot: HTSPLCTTNIKEGSNICLTRTDRGWYCDNAGSVSFFPQA P13843.1) DTCKVQSNRVFCDTMNSLTLPSEVSLCNTDIFNSKYDCK IMTSKTDISSSVITSLGAIVSCYGKTKCTASNKNRGIIKTF SNGCDYVSNKGVDTVSVGNTLYYVNKLEGKNLYVKGE PIINYYDPLVFPSDEFDASISQVNEKINQSLAFIRRSDELL HNVNTGKSTTNIMITTIIIVIIVVLLSLIAIGLLLYCKAKNT PVTLSKDQLSGINNIAFSK

    TABLE-US-00002 TABLE2 Aminoacidsequenceofwild-typeRSVFproteinbeforeartificialmutation Sequence nameand description SEQIDNo. Aminoacidsequence FproteinofRSV 7 QNITEEFYQSTCSAVSKGYLSALRTGWYTSVITIELSNIK strainA2 ENKCNGTDAKVKLIKQELDKYKNAVTELQLLMQSTPAT (SEQIDNo.1): NNRARRELPRFMNYTLNNAKKTNVTLSKKRKRRFLGFL anF0extracellular LGVGSAIASGVAVSKVLHLEGEVNKIKSALLSTNKAVVS region(withamino LSNGVSVLTSKVLDLKNYIDKQLLPIVNKQSCSISNIETVI acidsatpositions EFQQKNNRLLEITREFSVNAGVTTPVSTYMLTNSELLSLI 514to574removed NDMPITNDQKKLMSNNVQIVRQQSYSIMSIIKEEVLAYV ataC-terminal), VQLPLYGVIDTPCWKLHTSPLCTTNTKEGSNICLTRTDR andasignal GWYCDNAGSVSFFPQAETCKVQSNRVFCDTMNSLTLPS peptideremoved EVNLCNVDIFNPKYDCKIMTSKTDVSSSVITSLGAIVSCY GKTKCTASNKNRGIIKTFSNGCDYVSNKGVDTVSVGNT LYYVNKQEGKSLYVKGEPIINFYDPLVFPSDEFDASISQV NEKINQSLAFIRKSDELL FproteinofRSV 8 QNITEEFYQSTCSAVSKGYLSALRTGWYTSVITIELSNIK strainA2 ENKCNGTDAKVKLIKQELDKYKNAVTELQLLMQSTPAT (SEQIDNo.1): NNRARRELPRFMNYTLNNAKKTNVTLSKKRKRRFLGFL anF0extracellular LGVGSAIASGVAVSKVLHLEGEVNKIKSALLSTNKAVVS region(withamino LSNGVSVLTSKVLDLKNYIDKQLLPIVNKQSCSISNIETVI acidsatpositions EFQQKNNRLLEITREFSVNAGVTTPVSTYMLTNSELLSLI 530to574removed NDMPITNDQKKLMSNNVQIVRQQSYSIMSIIKEEVLAYV ataC-terminal), VQLPLYGVIDTPCWKLHTSPLCTTNTKEGSNICLTRTDR andasignal GWYCDNAGSVSFFPQAETCKVQSNRVFCDTMNSLTLPS peptideremoved EVNLCNVDIFNPKYDCKIMTSKTDVSSSVITSLGAIVSCY GKTKCTASNKNRGIIKTFSNGCDYVSNKGVDTVSVGNT LYYVNKQEGKSLYVKGEPIINFYDPLVFPSDEFDASISQV NEKINQSLAFIRKSDELLHNVNAGKSTTNIMITT FproteinofRSV 9 QNITEEFYQSTCSAVSKGYLSALRTGWYTSVITIELSNIK strainA2 ENKCNGTDAKVKLIKQELDKYKNAVTELQLLMQSTPAT (SEQIDNo.1): NNQARRELPRFMNYTLNNAKKTNVTLSKKRKSSFLGFL anF0extracellular LGVGSAIASGVAVSKVLHLEGEVNKIKSALLSTNKAVVS region(withamino LSNGVSVLTSKVLDLKNYIDKQLLPIVNKQSCSISNIETVI acidsatpositions EFQQKNNRLLEITREFSVNAGVTTPVSTYMLTNSELLSLI 514to574removed NDMPITNDQKKLMSNNVQIVRQQSYSIMSIIKEEVLAYV ataC-terminal), VQLPLYGVIDTPCWKLHTSPLCTTNTKEGSNICLTRTDR asignalpeptide GWYCDNAGSVSFFPQAETCKVQSNRVFCDTMNSLTLPS removed,mutation EVNLCNVDIFNPKYDCKIMTSKTDVSSSVITSLGAIVSCY ofacleavagesite, GKTKCTASNKNRGIIKTFSNGCDYVSNKGVDTVSVGNT andp27 LYYVNKQEGKSLYVKGEPIINFYDPLVFPSDEFDASISQV NEKINQSLAFIRKSDELL FproteinofRSV 10,11 F2polypeptide(SEQIDNo.10): strainA2 QNITEEFYQSTCSAVSKGYLSALRTGWYTSVITIELS (SEQIDNo.1): NIKENKCNGTDAKVKLIKQELDKYKNAVTELQLLMQST anF0extracellular PATNNRARR region(withamino F1polypeptide(SEQIDNo.11): acidsatpositions FLGFLLGVGSAIASGVAVSKVLHLEGEVNKIKSALLSTN 514to574removed KAVVSLSNGVSVLTSKVLDLKNYIDKQLLPIVNKQSCSIS ataC-terminal), NIETVIEFQQKNNRLLEITREFSVNAGVTTPVSTYMLINS asignalpeptide ELLSLINDMPITNDQKKLMSNNVQIVRQQSYSIMSIIKEE removed,a VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNTKEGSNICL combinationofan TRTDRGWYCDNAGSVSFFPQAETCKVQSNRVFCDTMN independentF2,and SLTLPSEVNLCNVDIFNPKYDCKIMTSKTDVSSSVITSLG p27artifically AIVSCYGKTKCTASNKNRGIIKTFSNGCDYVSNKGVDT removed VSVGNTLYYVNKQEGKSLYVKGEPIINFYDPLVFPSDEF DASISQVNEKINQSLAFIRKSDELL FproteinofRSV 12 QNITEEFYQSTCSAVSKGYLSALRTGWYTSVITIELSNIK strainA2 ENKCNGTDAKVKLIKQELDKYKNAVTELQLLMQSTPAT (SEQIDNo.1): NNQARGSGSGRSLGFLLGVGSAIASGVAVSKVLHLEGE anF0extracellular VNKIKSALLSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ region(withamino LLPIVNKQSCSISNIETVIEFQQKNNRLLEITREFSVNAGV acidsatpositions TTPVSTYMLTNSELLSLINDMPITNDQKKLMSNNVQIVR 514to574removed QQSYSIMSIIKEEVLAYVVQLPLYGVIDTPCWKLHTSPLC ataC-terminal), TTNTKEGSNICLTRTDRGWYCDNAGSVSFFPQAETCKV asignalpeptide QSNRVFCDTMNSLTLPSEVNLCNVDIFNPKYDCKIMTSK removed,F2-linker- TDVSSSVITSLGAIVSCYGKTKCTASNKNRGIIKTFSNGC F1,andalinker DYVSNKGVDTVSVGNTLYYVNKQEGKSLYVKGEPIINF replacingp27 YDPLVFPSDEFDASISQVNEKINQSLAFIRKSDELL FproteinofRSV 13 QNITEEFYQSTCSAVSKGYLSALRTGWYTSVITIELSNIK strainA2 ENKCNGTDAKVKLIKQELDKYKNAVTELQLLMQSTPAT (SEQIDNo.1): GSGSAIASGVAVSKVLHLEGEVNKIKSALLSTNKAVVSL anF0extracellular SNGVSVLTSKVLDLKNYIDKQLLPIVNKQSCSISNIETVIE region(withamino FQQKNNRLLEITREFSVNAGVTTPVSTYMLTNSELLSLIN acidsatpositions DMPITNDQKKLMSNNVQIVRQQSYSIMSIIKEEVLAYVV 514to574removed QLPLYGVIDTPCWKLHTSPLCTTNTKEGSNICLTRTDRG ataC-terminal), WYCDNAGSVSFFPQAETCKVQSNRVFCDTMNSLTLPSE asignalpeptide VNLCNVDIFNPKYDCKIMTSKTDVSSSVITSLGAIVSCY removed,F2-linker- GKTKCTASNKNRGIIKTFSNGCDYVSNKGVDTVSVGNT F1,alinker LYYVNKQEGKSLYVKGEPIINFYDPLVFPSDEFDASISQV replacingp27,and NEKINQSLAFIRKSDELL F2andF1withmore aminoacids removed FproteinofRSV 14 QNITEEFYQSTCSAVSKGYLSALRTGWYTSVITIELSNIK strainA2 ENKCNGTDAKVKLIKQELDKYKNAVTELQLLMQSTPPT (SEQIDNo.2): NNRARRELPRFMNYTLNNAKKTNVTLSKKRKRRFLGFL anF0extracellular LGVGSAIASGVAVSKVLHLEGEVNKIKSALLSTNKAVVS region(withamino LSNGVSVLTSKVLDLKNYIDKQLLPIVNKQSCSISNIETVI acidsatpositions EFQQKNNRLLEITREFSVNAGVTTPVSTYMLTNSELLSLI 514to574removed NDMPITNDQKKLMSNNVQIVRQQSYSIMSIIKEEVLAYV ataC-terminal), VQLPLYGVIDTPCWKLHTSPLCTTNTKEGSNICLTRTDR asignalpeptide GWYCDNAGSVSFFPQAETCKVQSNRVFCDTMNSLTLPS removed,ata EINLCNVDIFNPKYDCKIMTSKTDVSSSVITSLGAIVSCY C-terminal),anda GKTKCTASNKNRGIIKTFSNGCDYVSNKGMDTVSVGNT signalpeptide LYYVNKQEGKSLYVKGEPIINFYDPLVFPSDEFDASISQV removed NEKINQSLAFIRKSDELL FproteinofRSV 15 QNITEEFYQSTCSAVSKGYLSALRTGWYTSVITIELSNIK strainA2 ENKCNGTDAKVKLIKQELDKYKNAVTELQLLMQSTPPT (SEQIDNo.2): NNRARRELPRFMNYTLNNAKKTNVTLSKKRKRRFLGFL anF0extracellular LGVGSAIASGVAVSKVLHLEGEVNKIKSALLSTNKAVVS region(withamino LSNGVSVLTSKVLDLKNYIDKQLLPIVNKQSCSISNIETVI acidsatpositions EFQQKNNRLLEITREFSVNAGVTTPVSTYMLTNSELLSLI 514to574removed NDMPITNDQKKLMSNNVQIVRQQSYSIMSIIKEEVLAYV ataC-terminal), VQLPLYGVIDTPCWKLHTSPLCTTNTKEGSNICLTRTDR asignalpeptide GWYCDNAGSVSFFPQAETCKVQSNRVFCDTMNSLTLPS removed, EINLCNVDIFNPKYDCKIMTSKTDVSSSVITSLGAIVSCY GKTKCTASNKNRGIIKTFSNGCDYVSNKGMDTVSVGNT LYYVNKQEGKSLYVKGEPIINFYDPLVFPSDEFDASISQV NEKINQSLAFIRKSDELLHNVNAGKSTTNIMITT FproteinofRSV 16 QNITEEFYQSTCSAVSKGYLSALRTGWYTSVITIELSNIK strainA2 ENKCNGTDAKVKLIKQELDKYKNAVTELQLLMQSTPPT (SEQIDNo.2): NNQARRELPRFMNYTLNNAKKTNVTLSKKRKSSFLGFL anF0extracellular LGVGSAIASGVAVSKVLHLEGEVNKIKSALLSTNKAVVS region(withamino LSNGVSVLTSKVLDLKNYIDKQLLPIVNKQSCSISNIETVI acidsatpositions EFQQKNNRLLEITREFSVNAGVTTPVSTYMLTNSELLSLI 514to574removed NDMPITNDQKKLMSNNVQIVRQQSYSIMSIIKEEVLAYV ataC-terminal), VQLPLYGVIDTPCWKLHTSPLCTTNTKEGSNICLTRTDR asignalpeptide GWYCDNAGSVSFFPQAETCKVQSNRVFCDTMNSLTLPS removed,mutation EINLCNVDIFNPKYDCKIMTSKTDVSSSVITSLGAIVSCY ofacleavage GKTKCTASNKNRGIIKTFSNGCDYVSNKGMDTVSVGNT site,andp27 LYYVNKQEGKSLYVKGEPIINFYDPLVFPSDEFDASISQV NEKINQSLAFIRKSDELL FproteinofRSV 17,18 F2polypeptide(SEQIDNo.17): strainA2 QNITEEFYQSTCSAVSKGYLSALRTGWYTSVITIELSNIK (SEQIDNo.2): ENKCNGTDAKVKLIKQELDKYKNAVTELQLLMQSTPPT anF0extracellular NNRARR region(withamino F1polypeptide(SEQIDNo.18): acidsatpositions FLGFLLGVGSAIASGVAVSKVLHLEGEVNKIKSALLSTN 514to574removed KAVVSLSNGVSVLTSKVLDLKNYIDKQLLPIVNKQSCSIS ataC-terminal), NIETVIEFQQKNNRLLEITREFSVNAGVTTPVSTYMLINS asignalpeptide ELLSLINDMPITNDQKKLMSNNVQIVRQQSYSIMSIIKEE removed,a VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNTKEGSNICL combinationofan TRTDRGWYCDNAGSVSFFPQAETCKVQSNRVFCDTMN independentF1and SLTLPSEINLCNVDIFNPKYDCKIMTSKTDVSSSVITSLGA anindependentF2, IVSCYGKTKCTASNKNRGIIKTFSNGCDYVSNKGMDTV andp27artificially SVGNTLYYVNKQEGKSLYVKGEPIINFYDPLVFPSDEFD removed ASISQVNEKINQSLAFIRKSDELL FproteinofRSV 19 QNITEEFYQSTCSAVSKGYLSALRTGWYTSVITIELSNIK strainA2 ENKCNGTDAKVKLIKQELDKYKNAVTELQLLMQSTPPT (SEQIDNo.2): NNQARGSGSGRSLGFLLGVGSAIASGVAVSKVLHLEGE anF0extracellular VNKIKSALLSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ region(withamino LLPIVNKQSCSISNIETVIEFQQKNNRLLEITREFSVNAGV acidsatpositions TTPVSTYMLTNSELLSLINDMPITNDQKKLMSNNVQIVR 514to574removed QQSYSIMSIIKEEVLAYVVQLPLYGVIDTPCWKLHTSPLC ataC-terminal), TTNTKEGSNICLTRTDRGWYCDNAGSVSFFPQAETCKV asignalpeptide QSNRVFCDTMNSLTLPSEINLCNVDIFNPKYDCKIMTSK removed,F2-linker- TDVSSSVITSLGAIVSCYGKTKCTASNKNRGIIKTFSNGC F1,andalinker DYVSNKGMDTVSVGNTLYYVNKQEGKSLYVKGEPIINF replacingp27 YDPLVFPSDEFDASISQVNEKINQSLAFIRKSDELL FproteinofRSV 20 QNITEEFYQSTCSAVSKGYLSALRTGWYTSVITIELSNIK strainA2 ENKCNGTDAKVKLIKQELDKYKNAVTELQLLMQSTPPT (SEQIDNo.2): GSGSAIASGVAVSKVLHLEGEVNKIKSALLSTNKAVVSL anF0extracellular SNGVSVLTSKVLDLKNYIDKQLLPIVNKQSCSISNIETVIE region(withamino FQQKNNRLLEITREFSVNAGVTTPVSTYMLTNSELLSLIN acidsatpositions DMPITNDQKKLMSNNVQIVRQQSYSIMSIIKEEVLAYVV 514to574removed QLPLYGVIDTPCWKLHTSPLCTTNTKEGSNICLTRTDRG ataC-terminal), WYCDNAGSVSFFPQAETCKVQSNRVFCDTMNSLTLPSEI asignalpeptide NLCNVDIFNPKYDCKIMTSKTDVSSSVITSLGAIVSCYG removed,F2-linker- KTKCTASNKNRGIIKTFSNGCDYVSNKGMDTVSVGNTL F1,alinker YYVNKQEGKSLYVKGEPIINFYDPLVFPSDEFDASISQVN replacingF2andF1 EKINQSLAFIRKSDELL withmoreaminoacids removed FproteinofRSV 21 QNITEEFYQSTCSAVSKGYLSALRTGWYTSVITIELSNIK strainON1 ENKCNGTDAKVKLIKQELDKYKNAVTELQLLMQSTPAA (SEQIDNo.3): NSRARRELPRFMNYTLNNTKNTNVTLSKKRKRRFLGFL anF0extracellular LGVGSAIASGIAVSKVLHLEGEVNKIKSALLSTNKAVVS region(withamino LSNGVSVLTSKVLDLKNYIDKQLLPIVNKQSCSISNIETVI acidsatpositions EFQQKNNRLLEITREFSVNAGVTTPVSTYMLTNSELLSLI 514to574removed NDMPITNDQKKLMSSNVQIVRQQSYSIMSIIKEEVLAYV ataC-terminal), VQLPLYGVIDTPCWKLHTSPLCTTNTKEGSNICLTRTDR asignalpeptide GWYCDNAGSVSFFPQAETCKVQSNRVFCDTMNSLTLPS removed EVNLCNIDIFNPKYDCKIMTSKTDVSSSVITSLGAIVSCY GKTKCTASNKNRGIIKTFSNGCDYVSNKGVDTVSVGNT LYYVNKQEGKSLYVKGEPIINFYDPLVFPSDEFDASISQV NEKINQSLAFIRKSDELL FproteinofRSV 22 QNITEEFYQSTCSAVSKGYLSALRTGWYTSVITIELSNIK strainON1 ENKCNGTDAKVKLIKQELDKYKNAVTELQLLMQSTPAA (SEQIDNo.3): NSRARRELPRFMNYTLNNTKNTNVTLSKKRKRRFLGFL anF0extracellular LGVGSAIASGIAVSKVLHLEGEVNKIKSALLSTNKAVVS region(withamino LSNGVSVLTSKVLDLKNYIDKQLLPIVNKQSCSISNIETVI acidsatpositions EFQQKNNRLLEITREFSVNAGVTTPVSTYMLTNSELLSLI 530to574removed NDMPITNDQKKLMSSNVQIVRQQSYSIMSIIKEEVLAYV ataC-terminal), VQLPLYGVIDTPCWKLHTSPLCTTNTKEGSNICLTRTDR asignalpeptide GWYCDNAGSVSFFPQAETCKVQSNRVFCDTMNSLTLPS removed EVNLCNIDIFNPKYDCKIMTSKTDVSSSVITSLGAIVSCY GKTKCTASNKNRGIIKTFSNGCDYVSNKGVDTVSVGNT LYYVNKQEGKSLYVKGEPIINFYDPLVFPSDEFDASISQV NEKINQSLAFIRKSDELLHNVNAGKSTTNIMITT FproteinofRSV 23 QNITEEFYQSTCSAVSKGYLSALRTGWYTSVITIELSNIK strainON1 ENKCNGTDAKVKLIKQELDKYKNAVTELQLLMQSTPAA (SEQIDNo.3): NSQARRELPRFMNYTLNNTKNTNVTLSKKRKSSFLGFL anF0extracellular LGVGSAIASGIAVSKVLHLEGEVNKIKSALLSTNKAVVS region(withamino LSNGVSVLTSKVLDLKNYIDKQLLPIVNKQSCSISNIETVI acidsatpositions EFQQKNNRLLEITREFSVNAGVTTPVSTYMLTNSELLSLI 514to574removed NDMPITNDQKKLMSSNVQIVRQQSYSIMSIIKEEVLAYV ataC-terminal), VQLPLYGVIDTPCWKLHTSPLCTTNTKEGSNICLTRTDR asignalpeptide GWYCDNAGSVSFFPQAETCKVQSNRVFCDTMNSLTLPS removed,mutationof EVNLCNIDIFNPKYDCKIMTSKTDVSSSVITSLGAIVSCY acleavagesite, GKTKCTASNKNRGIIKTFSNGCDYVSNKGVDTVSVGNT andp27 LYYVNKQEGKSLYVKGEPIINFYDPLVFPSDEFDASISQV NEKINQSLAFIRKSDELL FproteinofRSV 24,25 F2polypeptide(SEQIDNo.24): strainON1 QNITEEFYQSTCSAVSKGYLSALRTGWYTSVITIELSNIK (SEQIDNo.3): ENKCNGTDAKVKLIKQELDKYKNAVTELQLLMQSTPAA anF0extracellular NSRARR region(withamino F1polypeptide(SEQIDNo.25): acidsatpositions FLGFLLGVGSAIASGIAVSKVLHLEGEVNKIKSALLSTNK 514to574removed AVVSLSNGVSVLTSKVLDLKNYIDKQLLPIVNKQSCSISN ataC-terminal), IETVIEFQQKNNRLLEITREFSVNAGVTTPVSTYMLINSE asignalpeptide LLSLINDMPITNDQKKLMSSNVQIVRQQSYSIMSIIKEEV removed,a LAYVVQLPLYGVIDTPCWKLHTSPLCTTNTKEGSNICLT combinationofan RTDRGWYCDNAGSVSFFPQAETCKVQSNRVFCDTMNSL independentF1and TLPSEVNLCNIDIFNPKYDCKIMTSKTDVSSSVITSLGAIV anindependentF2, SCYGKTKCTASNKNRGIIKTFSNGCDYVSNKGVDTVSV andp27artificially GNTLYYVNKQEGKSLYVKGEPIINFYDPLVFPSDEFDASI removed SQVNEKINQSLAFIRKSDELL FproteinofRSV 26 QNITEEFYQSTCSAVSKGYLSALRTGWYTSVITIELSNIK strainON1 ENKCNGTDAKVKLIKQELDKYKNAVTELQLLMQSTPAA (SEQIDNo.3): NSQARGSGSGRSLGFLLGVGSAIASGIAVSKVLHLEGEV anF0extracellular NKIKSALLSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQL region(withamino LPIVNKQSCSISNIETVIEFQQKNNRLLEITREFSVNAGVT acidsatpositions TPVSTYMLTNSELLSLINDMPITNDQKKLMSSNVQIVRQ 514to574removed QSYSIMSIIKEEVLAYVVQLPLYGVIDTPCWKLHTSPLCT ataC-terminal), TNTKEGSNICLTRTDRGWYCDNAGSVSFFPQAETCKVQ asignalpeptide SNRVFCDTMNSLTLPSEVNLCNIDIFNPKYDCKIMTSKT removed,F2-linker- DVSSSVITSLGAIVSCYGKTKCTASNKNRGIIKTFSNGCD F1,andalinker YVSNKGVDTVSVGNTLYYVNKQEGKSLYVKGEPIINFY replacingp27 DPLVFPSDEFDASISQVNEKINQSLAFIRKSDELL FproteinofRSV 27 QNITEEFYQSTCSAVSKGYLSALRTGWYTSVITIELSNIK strainON1 ENKCNGTDAKVKLIKQELDKYKNAVTELQLLMQSTPAA (SEQIDNo.3): GSGSAIASGIAVSKVLHLEGEVNKIKSALLSTNKAVVSLS anF0extracellular NGVSVLTSKVLDLKNYIDKQLLPIVNKQSCSISNIETVIEF region(withamino QQKNNRLLEITREFSVNAGVTTPVSTYMLTNSELLSLIN acidsatpositions DMPITNDQKKLMSSNVQIVRQQSYSIMSIIKEEVLAYVV 514to574removed QLPLYGVIDTPCWKLHTSPLCTTNTKEGSNICLTRTDRG ataC-terminal), WYCDNAGSVSFFPQAETCKVQSNRVFCDTMNSLTLPSE asignalpeptide VNLCNIDIFNPKYDCKIMTSKTDVSSSVITSLGAIVSCYG removed,F2-linker- KTKCTASNKNRGIIKTFSNGCDYVSNKGVDTVSVGNTL F1,alinker YYVNKQEGKSLYVKGEPIINFYDPLVFPSDEFDASISQVN replacingp27,andF2 EKINQSLAFIRKSDELL andF1withmore aminoacidsremoved FproteinofRSV 28 QNITEEFYQSTCSAVSRGYLSALRTGWYTSVITIELSNIK strainBAofsubtype ETKCNGTDTKVKLIKQELDKYKNAVTELQLLMQNTPAV B(SEQIDNo.4): NNRARREAPQYMNYTINTTKNLNVSISKKRKRRFLGFL anF0extracellular LGVGSAIASGIAVSKVLHLEGEVNKIKNALQLTNKAVVS region(withamino LSNGVSVLTSRVLDLKNYINNQLLPMVNRQSCRISNIET acidsatpositions VIEFQQKNSRLLEITREFSVNAGVTTPLSTYMLTNSELLS 514to574removed LINDMPITNDQKKLMSSNVQIVRQQSYSIMSIIKEEVLAY ataC-terminal),and VVQLPIYGVIDTPCWKLHTSPLCTTNIKEGSNICLTRTDR asignalpeptide GWYCDNAGSVSFFPQADTCKVQSNRVFCDTMNSLTLPS removed EVSLCNTDIFNSKYDCKIMTSKTDISSSVITSLGAIVSCYG KTKCTASNKNRGIIKTFSNGCDYVSNKGVDTVSVGNTL YYVNKLEGKNLYVKGEPIINYYDPLVFPSDEFDASISQV NEKINQSLAFIRRSDELL FproteinofRSV 29 QNITEEFYQSTCSAVSRGYLSALRTGWYTSVITIELSNIK strainBAofsubtype ETKCNGTDTKVKLIKQELDKYKNAVTELQLLMQNTPAV B(SEQIDNo.4): NNRARREAPQYMNYTINTTKNLNVSISKKRKRRFLGFL anF0extracellular LGVGSAIASGIAVSKVLHLEGEVNKIKNALQLTNKAVVS region(withamino LSNGVSVLTSRVLDLKNYINNQLLPMVNRQSCRISNIET acidsatpositions VIEFQQKNSRLLEITREFSVNAGVTTPLSTYMLTNSELLS 530to574removed LINDMPITNDQKKLMSSNVQIVRQQSYSIMSIIKEEVLAY ataC-terminal),and VVQLPIYGVIDTPCWKLHTSPLCTTNIKEGSNICLTRTDR asignalpeptide GWYCDNAGSVSFFPQADTCKVQSNRVFCDTMNSLTLPS removed EVSLCNTDIFNSKYDCKIMTSKTDISSSVITSLGAIVSCYG KTKCTASNKNRGIIKTFSNGCDYVSNKGVDTVSVGNTL YYVNKLEGKNLYVKGEPIINYYDPLVFPSDEFDASISQV NEKINQSLAFIRRSDELLHNVNTGKSTTNIMITA FproteinofRSV 30 QNITEEFYQSTCSAVSRGYLSALRTGWYTSVITIELSNIK strainBAofsubtype ETKCNGTDTKVKLIKQELDKYKNAVTELQLLMQNTPAV B(SEQIDNo.4): NNQARREAPQYMNYTINTTKNLNVSISKKRKSSFLGFLL anF0extracellular GVGSAIASGIAVSKVLHLEGEVNKIKNALQLTNKAVVSL region(withamino SNGVSVLTSRVLDLKNYINNQLLPMVNRQSCRISNIETVI acidsatpositions EFQQKNSRLLEITREFSVNAGVTTPLSTYMLTNSELLSLI 514to574removed NDMPITNDQKKLMSSNVQIVRQQSYSIMSIIKEEVLAYV ataC-terminal), VQLPIYGVIDTPCWKLHTSPLCTTNIKEGSNICLTRTDRG asignalpeptide WYCDNAGSVSFFPQADTCKVQSNRVFCDTMNSLTLPSE removed,mutationof VSLCNTDIFNSKYDCKIMTSKTDISSSVITSLGAIVSCYG acleavagesite, KTKCTASNKNRGIIKTFSNGCDYVSNKGVDTVSVGNTL andp27 YYVNKLEGKNLYVKGEPIINYYDPLVFPSDEFDASISQV NEKINQSLAFIRRSDELL FproteinofRSV 31,32 F2polypeptide(SEQIDNo.31): strainBAofsubtype QNITEEFYQSTCSAVSRGYLSALRTGWYTSVITIELSNIK B(SEQIDNo.4): ETKCNGTDTKVKLIKQELDKYKNAVTELQLLMQNTPAV anF0extracellular NNRARR region(withamino F1polypeptide(SEQIDNo.32): acidsatpositions FLGFLLGVGSAIASGIAVSKVLHLEGEVNKIKNALQLTN 514to574removed KAVVSLSNGVSVLTSRVLDLKNYINNQLLPMVNRQSCRI ataC-terminal), SNIETVIEFQQKNSRLLEITREFSVNAGVTTPLSTYMLIN asignalpeptide SELLSLINDMPITNDQKKLMSSNVQIVRQQSYSIMSIIKE removed,a EVLAYVVQLPIYGVIDTPCWKLHTSPLCTTNIKEGSNICL combinationofan TRTDRGWYCDNAGSVSFFPQADTCKVQSNRVFCDTMN independentF1an SLTLPSEVSLCNTDIFNSKYDCKIMTSKTDISSSVITSLGAI independentF2,and VSCYGKTKCTASNKNRGIIKTFSNGCDYVSNKGVDTVS p27artificially VGNTLYYVNKLEGKNLYVKGEPIINYYDPLVFPSDEFDA removed SISQVNEKINQSLAFIRRSDELL FproteinofRSV 33 QNITEEFYQSTCSAVSRGYLSALRTGWYTSVITIELSNIK strainBAofsubtype ETKCNGTDTKVKLIKQELDKYKNAVTELQLLMQNTPAV B(SEQIDNo.4): NNQARGSGSGRSLGFLLGVGSAIASGIAVSKVLHLEGEV anF0extracellular NKIKNALQLTNKAVVSLSNGVSVLTSRVLDLKNYINNQL region(withamino LPMVNRQSCRISNIETVIEFQQKNSRLLEITREFSVNAGV acidsatpositions TTPLSTYMLTNSELLSLINDMPITNDQKKLMSSNVQIVR 514to574removed QQSYSIMSIIKEEVLAYVVQLPIYGVIDTPCWKLHTSPLC ataC-terminal), TTNIKEGSNICLTRTDRGWYCDNAGSVSFFPQADTCKVQ asignalpeptide SNRVFCDTMNSLTLPSEVSLCNTDIFNSKYDCKIMTSKT removed,F2-linker- DISSSVITSLGAIVSCYGKTKCTASNKNRGIIKTFSNGCD F1,andalinker YVSNKGVDTVSVGNTLYYVNKLEGKNLYVKGEPIINYY replacingp27 DPLVFPSDEFDASISQVNEKINQSLAFIRRSDELL FproteinofRSV 34 QNITEEFYQSTCSAVSRGYLSALRTGWYTSVITIELSNIK strainBAofsubtype ETKCNGTDTKVKLIKQELDKYKNAVTELQLLMQNTPAV B(SEQIDNo.4): GSGSAIASGIAVSKVLHLEGEVNKIKNALQLTNKAVVSL anF0extracellular SNGVSVLTSRVLDLKNYINNQLLPMVNRQSCRISNIETVI region(withamino EFQQKNSRLLEITREFSVNAGVTTPLSTYMLTNSELLSLI acidsatpositions NDMPITNDQKKLMSSNVQIVRQQSYSIMSIIKEEVLAYV 514to574removed VQLPIYGVIDTPCWKLHTSPLCTTNIKEGSNICLTRTDRG ataC-terminal),and WYCDNAGSVSFFPQADTCKVQSNRVFCDTMNSLTLPSE asignalpeptide VSLCNTDIFNSKYDCKIMTSKTDISSSVITSLGAIVSCYG removed,F2-linker- KTKCTASNKNRGIIKTFSNGCDYVSNKGVDTVSVGNTL F1,alinker YYVNKLEGKNLYVKGEPIINYYDPLVFPSDEFDASISQV replacingp27,and NEKINQSLAFIRRSDELL F2andF1withmore aminoacidsremoved

    TABLE-US-00003 TABLE3 Illustrativeaminoacidsequenceofmutantofwild-typeRSVFprotein (afterartificialmutation) SEQ Mutant ID Introduced ID No. mutation Aminoacidsequence FHS-1 35 227N.fwdarw.M QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKMNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-2 36 227N.fwdarw.G QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKGNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-3 37 227N.fwdarw.A QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKANRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-4 38 227N.fwdarw.V QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKVNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-5 39 227N.fwdarw.L QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKLNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-6 40 227N.fwdarw.I QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKINRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-7 41 227N.fwdarw.W QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKWNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-8 42 227N.fwdarw.F QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKFNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-9 43 227N.fwdarw.P QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKPNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-10 44 227N.fwdarw.S QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKSNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-11 45 227N.fwdarw.T QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKTNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-12 46 227N.fwdarw.C QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKCNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-13 47 227N.fwdarw.Y QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKYNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-14 48 227N.fwdarw.Q QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKQNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-15 49 227N.fwdarw.D QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKDNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-16 50 227N.fwdarw.E QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKENRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-17 51 227N.fwdarw.K QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKKNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-18 52 227N.fwdarw.R QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKRNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-19 53 227N.fwdarw.H QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKHNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-20 54 230L.fwdarw.M QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRMLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-21 55 230L.fwdarw.G QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRGLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-22 56 230L.fwdarw.A QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRALEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVG NTLYYVNKQEGKSLYVKGEPIINFYDPLVFP SDEFDASISQVNEKINQSLAFIRKSDELL FHS-23 57 230L.fwdarw.V QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRVLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-24 58 230L.fwdarw.I QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRILEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-25 59 230L.fwdarw.W QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRWLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-26 60 230L.fwdarw.F QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRFLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-27 61 230L.fwdarw.P QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRPLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-28 62 230L.fwdarw.S QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRSLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-29 63 230L.fwdarw.T QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRTLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-30 64 230L.fwdarw.C QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRCLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-31 65 230L.fwdarw.Y QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRYLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-32 66 230L.fwdarw.Q QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRQLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-33 67 230L.fwdarw.N QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRNLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-34 68 230L.fwdarw.D QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRDLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-35 69 230L.fwdarw.E QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRELEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-36 70 230L.fwdarw.K QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRKLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-37 71 230L.fwdarw.R QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRRLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-38 72 230L.fwdarw.H QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRHLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-39 73 234T.fwdarw.L QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI LREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-40 74 234T.fwdarw.I QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI IREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-41 75 234T.fwdarw.M QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI MREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-42 76 234T.fwdarw.F QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI FREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-43 77 234T.fwdarw.Y QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI YREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-44 78 234T.fwdarw.W QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI WREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-45 79 238S.fwdarw.L QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFLVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-46 80 238S.fwdarw.I QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFIVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-47 81 238S.fwdarw.M QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFMVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-48 82 238S.fwdarw.F QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFFVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-49 83 238S.fwdarw.Y QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFYVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-50 84 238S.fwdarw.W QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFWVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-51 85 82E.fwdarw.L QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQLLDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-52 86 82E.fwdarw.I QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQILDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-53 87 82E.fwdarw.M QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQMLDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-54 88 82E.fwdarw.F QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQFLDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-55 89 82E.fwdarw.Y QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQYLDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-56 90 82E.fwdarw.W QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQWLDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-57 91 86F.fwdarw.W QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKWK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-58 92 89A.fwdarw.L QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NLVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-59 93 89A.fwdarw.I QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NIVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-60 94 89A.fwdarw.M QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NMVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-61 95 89A.fwdarw.F QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NFVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-62 96 89A.fwdarw.Y QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NYVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-63 97 89A.fwdarw.W QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NWVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-64 98 92E.fwdarw.L QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTLLQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-65 99 92E.fwdarw.I QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTILQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-66 100 92E.fwdarw.M QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTMLQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-67 101 92E.fwdarw.F QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTFLQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-68 102 92E.fwdarw.Y QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTYLQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-69 103 92E.fwdarw.W QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTWLQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-70 104 93L.fwdarw.M QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTEMQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-71 105 93L.fwdarw.F QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTEFQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-72 106 93L.fwdarw.Y QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTEYQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-73 107 93L.fwdarw.W QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTEWQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-74 108 96L.fwdarw.M QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLMMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-75 109 96L.fwdarw.F QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLFMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-76 110 96L.fwdarw.Y QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLYMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-77 111 96L.fwdarw.W QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLWMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-78 112 193L.fwdarw.M QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVMDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-79 113 193L.fwdarw.F QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVFDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-80 114 193L.fwdarw.Y QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVYDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-81 115 193L.fwdarw.W QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVWDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-82 116 195L.fwdarw.M QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDMKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-83 117 195L.fwdarw.F QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDFKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-84 118 195L.fwdarw.Y QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDYKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-85 119 195L.fwdarw.W QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDWKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-86 120 199I.fwdarw.M QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYMDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-87 121 199I.fwdarw.F QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYFDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-88 122 199I.fwdarw.Y QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYYDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-89 123 199I.fwdarw.W QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYWDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-90 124 190S.fwdarw.V QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTVKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-91 125 141L.fwdarw.M QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF MLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-92 126 141L.fwdarw.F QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF FLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-93 127 141L.fwdarw.Y QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF YLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-94 128 141L.fwdarw.W QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF WLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-95 129 373L.fwdarw.M QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSMTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-96 130 373L.fwdarw.F QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSFTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-97 131 373L.fwdarw.Y QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSYTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHS-98 132 373L.fwdarw.W QNITEEFYQSTCSAVSKGYLSALRTGWYTSV ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSWTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHM-1 133 227N.fwdarw.F, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 230L.fwdarw.F ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKFNRFLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHM-2 134 227N.fwdarw.M, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 82E.fwdarw.L ITIELSNIKENKCNGTDAKVKLIKQLLDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKMNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHM-3 135 227N.fwdarw.M, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 89A.fwdarw.L ITIELSNIKENKCNGTDAKVKLIKQELDKYK NLVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKMNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHM-4 136 227N.fwdarw.M, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 93L.fwdarw.F ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTEFQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKMNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHM-5 137 227N.fwdarw.M, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 96L.fwdarw.M ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLMMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKMNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHM-6 138 230L.fwdarw.F, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 89A.fwdarw.F ITIELSNIKENKCNGTDAKVKLIKQELDKYK NFVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRFLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHM-7 139 230L.fwdarw.F, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 82E.fwdarw.F ITIELSNIKENKCNGTDAKVKLIKQFLDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRFLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHM-8 140 230L.fwdarw.F, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 93L.fwdarw.F ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTEFQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRFLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHM-9 141 230L.fwdarw.F, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 238S.fwdarw.M ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRFLEI TREFMVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHM-10 142 227N.fwdarw.M, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 234T.fwdarw.F ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKMNRLLEI FREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHM-11 143 82E.fwdarw.F, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 234T.fwdarw.F ITIELSNIKENKCNGTDAKVKLIKQFLDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI FREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHM-12 144 193L.fwdarw.M, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 195L.fwdarw.M ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVMDMKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHM-13 145 190S.fwdarw.V, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 193L.fwdarw.M, ITIELSNIKENKCNGTDAKVKLIKQELDKYK 195L.fwdarw.M NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTVKVMDMKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHM-14 146 193L.fwdarw.M, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 199I.fwdarw.M ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVMDLKNYMDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHM-15 147 193L.fwdarw.M, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 199I.fwdarw.F ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVMDLKNYFDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHM-16 148 141L.fwdarw.M, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 373L.fwdarw.M ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF MLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSMTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHM-17 149 141L.fwdarw.F, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 373L.fwdarw.F ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF FLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSFTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHM-18 150 82E.fwdarw.C, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 224Q.fwdarw.C ITIELSNIKENKCNGTDAKVKLIKQCLDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFCQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHM-19 151 92E.fwdarw.C, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 238S.fwdarw.C ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTCLQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFCVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHM-20 152 79I.fwdarw.C, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 220V.fwdarw.C ITIELSNIKENKCNGTDAKVKLCKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETCIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHM-21 153 97M.fwdarw.C, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 2911.fwdarw.C ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLCQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSCIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHM-22 154 93L.fwdarw.C, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 234T.fwdarw.C ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTECQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI CREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHM-23 155 92E.fwdarw.C, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 234T.fwdarw.C ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTCLQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKNNRLLEI CREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHM-24 156 227N.fwdarw.M, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 230L.fwdarw.F ITIELSNIKENKCNGTDAKVKLIKQELDKYK NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKMNRFLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHM-25 157 190S.fwdarw.V, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 193L.fwdarw.M, ITIELSNIKENKCNGTDAKVKLIKQELDKYK 227N.fwdarw.M, NAVTELQLLMQSTPATNNQARGSGSGRSLGF 230L.fwdarw.F LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTVKVMDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKMNRFLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKSDEL L FHCS-1 158 227N.fwdarw.M, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 230L.fwdarw.F, ITIELSNIKENKCNGTDAKVKLIKQELDKYK 509S.fwdarw.M NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKMNRFLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKMDEL L FHCS-2 159 227N.fwdarw.M, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 230L.fwdarw.F, ITIELSNIKENKCNGTDAKVKLIKQELDKYK 509S.fwdarw.F NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKMNRFLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKFDEL L FHCS-3 160 227N.fwdarw.M, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 230L.fwdarw.F, ITIELSNIKENKCNGTDAKVKLIKQELDKYK 509S.fwdarw.L NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKMNRFLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAFIRKLDEL L FHCS-4 161 227N.fwdarw.M, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 230L.fwdarw.F, ITIELSNIKENKCNGTDAKVKLIKQELDKYK 505F.fwdarw.W NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKMNRFLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAWIRKSDEL L FHCS-5 162 227N.fwdarw.M, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 230L.fwdarw.F, ITIELSNIKENKCNGTDAKVKLIKQELDKYK 502S.fwdarw.Y NAVTELQLLMQSTPATNNQARGSGSGRSLGF LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKMNRFLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLSEVNLCNVDIFNP KYDCKIMTSKTDVSSSVITSLGAIVSCYGKT KCTASNKNRGIIKTFSNGCDYVSNKGVDTVS VGNTLYYVNKQEGKSLYVKGEPIINFYDPLV FPSDEFDASISQVNEKINQYLAFIRKSDELL FHCM-1 163 227N.fwdarw.M, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 230L.fwdarw.F, ITIELSNIKENKCNGTDAKVKLIKQELDKYK 505F.fwdarw.W, NAVTELQLLMQSTPATNNQARGSGSGRSLGF 509S.fwdarw.L LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKMNRFLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAWIRKLDEL L FHCM-2 164 227N.fwdarw.M, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 230L.fwdarw.F, ITIELSNIKENKCNGTDAKVKLIKQELDKYK 501Q.fwdarw.M, NAVTELQLLMQSTPATNNQARGSGSGRSLGF 509S.fwdarw.F LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL LSTNKAVVSLSNGVSVLTSKVLDLKNYIDKQ LLPIVNKQSCSISNIETVIEFQQKMNRFLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINMSLAFIRKFDEL L FHCM-3 165 227N.fwdarw.M, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 230L.fwdarw.F, ITIELSNIKENKCNGTDAKVKLIKQELDKYK 190S.fwdarw.V, NAVTELQLLMQSTPATNNQARGSGSGRSLGF 193L.fwdarw.M, LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL 505F.fwdarw.W, LSTNKAVVSLSNGVSVLTVKVMDLKNYIDKQ 509S.fwdarw.L LLPIVNKQSCSISNIETVIEFQQKMNRFLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINQSLAWIRKLDEL L FHCM-4 166 227N.fwdarw.M, QNITEEFYQSTCSAVSKGYLSALRTGWYTSV 230L.fwdarw.F, ITIELSNIKENKCNGTDAKVKLIKQELDKYK 190S.fwdarw.V, NAVTELQLLMQSTPATNNQARGSGSGRSLGF 193L.fwdarw.M, LLGVGSAIASGVAVSKVLHLEGEVNKIKSAL 501Q.fwdarw.M, LSTNKAVVSLSNGVSVLTVKVMDLKNYIDKQ 509S.fwdarw.F LLPIVNKQSCSISNIETVIEFQQKMNRFLEI TREFSVNAGVTTPVSTYMLTNSELLSLINDM PITNDQKKLMSNNVQIVRQQSYSIMSIIKEE VLAYVVQLPLYGVIDTPCWKLHTSPLCTTNT KEGSNICLTRTDRGWYCDNAGSVSFFPQAET CKVQSNRVFCDTMNSLTLPSEVNLCNVDIFN PKYDCKIMTSKTDVSSSVITSLGAIVSCYGK TKCTASNKNRGIIKTFSNGCDYVSNKGVDTV SVGNTLYYVNKQEGKSLYVKGEPIINFYDPL VFPSDEFDASISQVNEKINMSLAFIRKFDEL L

    DETAILED DESCRIPTION OF THE EMBODIMENTS

    [0293] A mutant of a respiratory syncytial virus (RSV) fusion (F) protein is disclosed in the present disclosure. Those skilled in the art can refer to contents herein to appropriately improve process parameters for implementation. It is specifically noted that all similar substitutions and modifications will be apparent to those skilled in the art and are deemed to be included in the present disclosure. It will be apparent to those skilled in the art that modifications or suitable variations and combinations of what is described herein can be made to practice and apply the disclosed technology without departing from the content, spirit and scope of the present disclosure.

    [0294] In the present disclosure, scientific and technical terms used herein have the meanings commonly understood by those skilled in the art, unless otherwise specified. Definitions of common terms in molecular biology can be found in Lewin's GENES XII, Jocelyn E. Krebs/Elliott S. Goldstein/Stephen T. Kilpatrick, by Jones & Bartlett Publishers, 2018.

    [0295] Throughout the specification and claims, unless explicitly indicated otherwise, the term comprise, or its variations of include or contain, and the like, will be understood to include stated elements or components but not to exclude other elements or components. The terms a, an, and the include plural designators. The term plurality means two or more. The terms such as, for example, and the like are intended to refer to illustrative embodiments and are not intended to limit the scope of the present disclosure.

    [0296] In order to enable those skilled in the art to better understand the technical solutions of the present disclosure, the present disclosure is described in further detail below in combination with specific examples.

    I. DEFINITION

    [0297] The term wild-type, which may also be referred to as natural, native, refers to mutation that does not occur unnaturally. In some embodiments, illustrative instances of a wild-type RSV F protein are, for example, sequences shown in SEQ ID No. 1-6.

    [0298] The term mutation refers to a change in a sequence (for example, nucleotide or amino acid sequence) relative to a corresponding sequence in a native, wild-type, standard, or reference form (that is, a non-mutated sequence). Amino acid mutation generally includes a substitution, a deletion or an insertion of an amino acid residue. The mutation can be artificial or naturally formed.

    [0299] The terms peptide, polypeptide and protein are used interchangeably and generally refer to peptides and proteins with amino acids covalently linked by peptide bonds. The term protein encompasses purified natural products, or products that may be produced partially or wholly by using recombinant or synthetic techniques. The terms peptide and protein may refer to aggregates of proteins, such as dimers or other multimers, fusion proteins, protein variants, or their derivatives. The terms also include proteins with modifications, for example, proteins modified by glycosylation, acetylation, phosphorylation, PEGylation, and ubiquitination. A protein may include amino acids that are not encoded by a codon of a nucleic acid. A protein may have an amino acid sequence with a sufficient length to produce of a tertiary and/or quaternary structure with higher levels.

    [0300] The term cleavage site for a protease refers to a specific amino acid sequence motif within amino acids of a protein or a fusion protein. Within an amino acid sequence of a protein or a fusion protein, the protein or the fusion protein is cleaved by a specific protease that recognizes the amino acid sequence. In some embodiments, instances of the specific protease are, for example, furin, trypsin, factor Xa, thrombin, or cathepsin L.

    [0301] The term F0 precursor protein refers to that an RSV F protein is first translated in vivo into a single chain inactive precursor protein F0 consisting of 574 amino acids. A signal peptide consisting of amino acids at positions 1 to 25 at an N-terminus of the F0 protein is removed by a signal peptidase. Furthermore, there are two furin cleavage sites at amino acids at positions 109/110 and 136/137 of the F0. Through the protease cleavage, a polypeptide with amino acids at positions 110 to 136 (referred to as p27) is removed, and an F1 consisting of amino acids at positions 137 to 574 and an F2 consisting of amino acids at positions 26 to 109 are generated.

    [0302] The terms head domain and tail domain are defined in order to describe the structure of the RSV F protein visually in the present disclosure. The amino acids at positions 1 to 485 are defined as the head domain. The amino acid at position 486 to the C-terminus of the F protein are defined as the tail domain. The approximate spatial positions of the head domain and the tail domain are shown in FIG. 1A.

    [0303] The term virus-like particles (VLPs) or pseudoviruses refers to a polyprotein structures composed of corresponding native viral structural proteins, but lacking the entire viral genome or part of the viral genome, particularly replicative and infectious components of the viral genome. Thus the virus-like particles are not replicative and infectious. The polyprotein structure closely mimics its corresponding native viral particles in morphology and size and can be spontaneously formed upon recombinant expression of the viral structural proteins.

    [0304] The term protein delivery vector refers to a protein that can act as a carrier for an epitope peptide, that is, it can insert an epitope peptide into a specific position (for example, inside the protein, an N-terminal or a C-terminal), such that the epitope peptide can be presented and thus recognized by antibodies or an immune system.

    [0305] The term immunogenicity refers to an ability of a substance to elicit, cause, stimulate or induce an immune response in an animal against a particular antigen, in the presence or absence of an adjuvant.

    [0306] The term conjugate generally refers to any substance formed by joining together two or more independent moieties. For example, a conjugate may include a substance in which one polypeptide is linked to another polypeptide or polypeptides. When part of the conjugate is a protein, the conjugate is a protein conjugate.

    II. MUTATION DESIGN OF A HEAD DOMAIN OF AN RSV F PROTEIN STABILIZES THE F PROTEIN IN A PREFUSION TRIMER STATE

    [0307] It has been reported that most of highly active neutralizing antibodies against the RSV F protein target a prefusion conformation (Ref: Battles M. B., McLellan J. S. Nat. Rev. Microbiol., 2019, 17:233-245; Gilman M. S. A. et al. PLoS Pathog., 2015, 11:e1005035; Harshbarger W., et al. MAbs, 2021, 13:1955812.), and a neutralizing antibody titer induced by an F protein trimer in a prefusion conformation is much higher than that induced by an F protein trimer in a postfusion conformation (Ref: Battles M. B., McLellan J. S. Nat. Rev. Microbiol., 2019, 17:233-245; Krarup A. et al. Nat. Commun., 2015, 6:8143), such that the F protein trimer in the prefusion conformation is a preferred antigen for development of recombinant subunit vaccines. However, the F protein trimer in the prefusion conformation is in a metastable state and is prone to a transition from the prefusion conformation (shown in FIG. 1A) to the postfusion conformation (shown in FIG. 1B). One of key issues in the development of the recombinant subunit vaccines is to stabilize the F protein in a prefusion trimer state through modification of a key site.

    [0308] Spatial structures of a prefusion RSV F protein trimer and a postfusion RSV F protein trimer are obtained by X-ray crystal structure analysis (Ref: McLellan J. S., et al. Science, 2013, 340:1113-1117. McLellan J. S., et al. J. Virol., 2011, 85:7788-7796.), and submitted to a protein data bank (PDB). Spatial structure coordinates of the prefusion trimer and the postfusion trimer can be downloaded from the PDB according to PDB identifiers 4jhw and 3rrr. By using commonly used protein structure alignment software, such as Pymol and Chimera, the prefusion conformation and the postfusion conformation are subjected to structural superposition, as shown in FIG. 2. As can be seen from a structural alignment result of FIG. 2, from the prefusion conformation to the postfusion conformation, RR1 of the head domain and RR2 located at the C-terminal of the head domain and the tail domain undergo a large conformational change. In this case, 1 and 5 of the head domain also correspondingly undergo obvious conformational movement. Thus conformational changes of 1 and 5 are coupled with a conformational change of RR1. Based on the above structure analysis, stability mutation is introduced into an 1 or 5 region to prevent the conformational changes of 1 and 5. The conformational change of RR1 is prevented through coupling regulation. Thus the F protein can be stabilized in a prefusion conformation state. Furthermore, stability mutation can alternatively be introduced directly into an interaction region involved in RR1 to prevent the transition from the prefusion conformation to the postfusion conformation. For this reason, the present disclosure introduces mutation into an 1 helix (amino acids at positions 73 to 96) or an 5 helix (amino acids at positions 216 to 240) or directly into the interaction region involved in RR1, or any combination of the mutation. Interactions between residues are enhanced to stabilize conformations of 1, 5, or RR1, and thus the F protein is stabilized in the prefusion trimer state. Some illustrative single-site mutation design is shown in Table 4. The single-site mutation can be combined arbitrarily and can have similar or stronger stabilizing effects. Some illustrative mutation combinations are shown in Table 5.

    TABLE-US-00004 TABLE 4 Illustrative single-site mutation design for head domain of RSV F protein Mutant Introduced Mutant Introduced number mutation number mutation FHS-1 227N.fwdarw.M FHS-2 227N.fwdarw.G FHS-3 227N.fwdarw.A FHS-4 227N.fwdarw.V FHS-5 227N.fwdarw.L FHS-6 227N.fwdarw.I FHS-7 227N.fwdarw.W FHS-8 227N.fwdarw.F FHS-9 227N.fwdarw.P FHS-10 227N.fwdarw.S FHS-11 227N.fwdarw.T FHS-12 227N.fwdarw.C FHS-13 227N.fwdarw.Y FHS-14 227N.fwdarw.Q FHS-15 227N.fwdarw.D FHS-16 227N.fwdarw.E FHS-17 227N.fwdarw.K FHS-18 227N.fwdarw.R FHS-19 227N.fwdarw.H FHS-20 230L.fwdarw.M FHS-21 230L.fwdarw.G FHS-22 230L.fwdarw.A FHS-23 230L.fwdarw.V FHS-24 230L.fwdarw.I FHS-25 230L.fwdarw.W FHS-26 230L.fwdarw.F FHS-27 230L.fwdarw.P FHS-28 230L.fwdarw.S FHS-29 230L.fwdarw.T FHS-30 230L.fwdarw.C FHS-31 230L.fwdarw.Y FHS-32 230L.fwdarw.Q FHS-33 230L.fwdarw.N FHS-34 230L.fwdarw.D FHS-35 230L.fwdarw.E FHS-36 230L.fwdarw.K FHS-37 230L.fwdarw.R FHS-38 230L.fwdarw.H FHS-39 234T.fwdarw.L FHS-40 234T.fwdarw.I FHS-41 234T.fwdarw.M FHS-42 234T.fwdarw.F FHS-43 234T.fwdarw.Y FHS-44 234T.fwdarw.W FHS-45 238S.fwdarw.L FHS-46 238S.fwdarw.I FHS-47 238S.fwdarw.M FHS-48 238S.fwdarw.F FHS-49 238S.fwdarw.Y FHS-50 238S.fwdarw.W FHS-51 82E.fwdarw.L FHS-52 82E.fwdarw.I FHS-53 82E.fwdarw.M FHS-54 82E.fwdarw.F FHS-55 82E.fwdarw.Y FHS-56 82E.fwdarw.W FHS-57 86F.fwdarw.W FHS-58 89A.fwdarw.L FHS-59 89A.fwdarw.I FHS-60 89A.fwdarw.M FHS-61 89A.fwdarw.F FHS-62 89A.fwdarw.Y FHS-63 89A.fwdarw.W FHS-64 92E.fwdarw.L FHS-65 92E.fwdarw.I FHS-66 92E.fwdarw.M FHS-67 92E.fwdarw.F FHS-68 92E.fwdarw.Y FHS-69 92E.fwdarw.W FHS-70 93L.fwdarw.M FHS-71 93L.fwdarw.F FHS-72 93L.fwdarw.Y FHS-73 93L.fwdarw.W FHS-74 96L.fwdarw.M FHS-75 96L.fwdarw.F FHS-76 96L.fwdarw.Y FHS-77 96L.fwdarw.W FHS-78 193L.fwdarw.M FHS-79 193L.fwdarw.F FHS-80 193L.fwdarw.Y FHS-81 193L.fwdarw.W FHS-82 195L.fwdarw.M FHS-83 195L.fwdarw.F FHS-84 195L.fwdarw.Y FHS-85 195L.fwdarw.W FHS-86 199I.fwdarw.M FHS-87 199I.fwdarw.F FHS-88 199I.fwdarw.Y FHS-89 199I.fwdarw.W FHS-90 190S.fwdarw.V FHS-91 141L.fwdarw.M FHS-92 141L.fwdarw.F FHS-93 141L.fwdarw.Y FHS-94 141L.fwdarw.W FHS-95 373L.fwdarw.M FHS-96 373L.fwdarw.F FHS-97 373L.fwdarw.Y FHS-98 373L.fwdarw.W

    TABLE-US-00005 TABLE 5 Illustrative site mutation combination design for head domain of RSV F protein Mutant Introduced number mutation FHM-1 227N.fwdarw.F, 230L.fwdarw.F FHM-2 227N.fwdarw.M, 82E.fwdarw.L FHM-3 227N.fwdarw.M, 89A.fwdarw.L FHM-4 227N.fwdarw.M, 93L.fwdarw.F FHM-5 227N.fwdarw.M, 96L.fwdarw.M FHM-6 230L.fwdarw.F, 89A.fwdarw.F FHM-7 230L.fwdarw.F, 82E.fwdarw.F FHM-8 230L.fwdarw.F, 93L.fwdarw.F FHM-9 230L.fwdarw.F, 238S.fwdarw.M FHM-10 227N.fwdarw.M, 234T.fwdarw.F FHM-11 82E.fwdarw.F, 234T.fwdarw.F FHM-12 193L.fwdarw.M, 195L.fwdarw.M FHM-13 190S.fwdarw.V, 193L.fwdarw.M, 195L.fwdarw.M FHM-14 193L.fwdarw.M, 199I.fwdarw.M FHM-15 193L.fwdarw.M, 199I.fwdarw.F FHM-16 141L.fwdarw.M, 373L.fwdarw.M FHM-17 141L.fwdarw.F, 373L.fwdarw.F FHM-18 82E.fwdarw.C, 224Q.fwdarw.C FHM-19 92E.fwdarw.C, 238S.fwdarw.C FHM-20 79I.fwdarw.C, 220V.fwdarw.C FHM-21 97M.fwdarw.C, 291I.fwdarw.C FHM-22 93L.fwdarw.C, 234T.fwdarw.C FHM-23 92E.fwdarw.C, 234T.fwdarw.C FHM-24 227N.fwdarw.M, 230L.fwdarw.F FHM-25 227N.fwdarw.M, 230L.fwdarw.F, 190S.fwdarw.V, 193L.fwdarw.M

    III. EXAMPLE

    Example 1 Plasmid Construction, Recombinant Expression and Effect Validation of a Mutant Designed by Introducing Artificial Mutation into a Head Domain (Table 4 and Table 5)

    [0309] A natural RSV F protein is a homotrimer. In order to facilitate secretion expression of a protein in the example, amino acids at positions 514 to 574 including a transmembrane region and an intracellular region were removed from an F0 full-length sequence. Such a region plays an important role in formation of an F protein trimer. In order to assemble the cut sequence to form a trimer, a C-terminal of the cut sequence was linked to an exogenous trimerization domain (a T4 Foldon trimerization motif was used in the example) via a short linker (with a sequence being SAIG). Some studies showed that removing a sequence of p27 from an F0 contributed to trimer formation (Krarup A. et al. Nat. Commun., 2015, 6:8143.). In the example, a sequence (RARRELPRFMNYTLNNAKKTNVTLSKKRKRRF) at positions 107 to 138 containing p27 and a furin cleavage site was replaced by a linker QARGSGSGRS sequence. Moreover, a signal peptide MELLILKANAITTILTAVTFCFASG composed of amino acids at positions 1 to 25 at an N-terminal of the sequence was replaced by a signal peptide sequence MDMRVPAQLLGLLLLWLRGARC suitable for a recombinant expression system HEK293, so as to promote secretion expression of a recombinant protein. On this basis, corresponding mutants of the RSV F protein were constructed according to site mutation of the head domain designed in Table 4 and Table 5. Amino acid sequences of the constructed mutants (excluding the signal peptide, T4 foldon and a linker) were shown in SEQ ID No. 35-157.

    [0310] According to the amino acid sequences shown in SEQ ID No. 35-157, the signal peptide suitable for the recombinant expression system HEK293, the T4 foldon trimerization motif, the linker, and an additional tag sequence useful for purification were added. Codon optimization was performed according to a codon bias of the HEK293 to form corresponding coding gene sequences. The coding gene sequences were constructed in a whole gene synthesis or point mutation mode. A Kozak sequence GCCACC was added at a front end of each sequence and then connected to a recombinant expression vector pcDNA3.1 to form a recombinant expression plasmid. An Expi293 transient expression system (Thermo Fisher) was used to perform recombinant expression on the constructed mutants of the prefusion RSV F protein, and cell secretion supernatant was collected on the 5th day after transfection. As a control, a wild-type RSV F protein before artificial mutation (referred to as wild-type RSV F protein 1 before artificial mutation) was subjected recombinant expression according to SEQ ID No. 12 based on the above method. Moreover, an RSV strain A2 F protein (Catalog Number: 11049-V08B) was purchased from Sino Biological, Inc. as another control (referred to as wild-type RSV F protein 2 before artificial mutation), whose amino acid sequence was shown in SEQ ID No. 15.

    [0311] In order to verify whether introduction of the above mutation can stabilize the RSV F protein in a prefusion trimer state, the example used a prefusion trimer-specifically binding monoclonal antibody AM14 to detect binding activity of the above constructed mutants with the AM14 antibody through an enzyme-linked immunosorbent assay (ELISA) experimental method. The binding activity of the constructed mutants was compared with AM14 monoclonal antibody binding activity of the wild-type RSV F protein 1 before artificial mutation and the wild-type RSV F protein 2 before artificial mutation. Published literature showed that the AM14 antibody binded to an epitope region at an interface between two adjacent monomers in the prefusion F protein trimer (Ref: Gilman M. S. A. et al. PLoS Pathog., 2015, 11:e1005035; Harshbarger W., et al. MAbs, 2021, 13:1955812.), and the epitope was disrupted when the F protein trimer transitions from the prefusion conformation to the postfusion conformation. Thus the AM14 antibody cannot bind to the postfusion F protein. The AM14 antibody was typically used for specific recognition and detection of a prefusion F protein trimer structure (Ref: Gilman M. S. A. et al. PLoS Pathog., 2015, 11:e1005035; Harshbarger W., et al. MAbs, 2021, 13:1955812.). Furthermore, it has been reported that the AM14 monoclonal antibody binding activity was positively correlated with a neutralizing antibody titer produced by immunized animals (Ref: Che Y. et al. Sci. Transl. Med., 2023, 15:eade6422.). Moreover, in order to verify whether the constructed mutants can be expressed, a His-tag antibody was used to detect expression of the mutants in the example.

    [0312] An ELISA process of the transfection supernatant was as follows:

    (1) Assay of Binding Activity of AM14 Monoclonal Antibody

    [0313] The AM14 monoclonal antibody (Wuhan Chemstan Biotechnology Co., Ltd.) was diluted to 1 g/ml by using a coating buffer (Na.sub.2CO.sub.3NaHCO.sub.3 solution, pH9.6) and coated a 96-well ELISA plate at 100 l/well at 4 C. for 8 h to 12 h. The plate was washed by phosphate buffered saline with tween (PBST). A blocking buffer (20% calf serum, 0.2% yeast extract, the PBST was dissolved) was added at 100 l/well and blocked at 37 C. for 2 h. The plate was washed by the PBST. 10-fold diluted cell secretion supernatant of centrifuged recombinant expression mutants (including the wild-type RSV F protein 1 before artificial mutation) and 2 g/ml wild-type RSV F protein 2 before artificial mutation were added at 100 l/well and incubated at 37 C. for 1 h. The plate was washed by the PBST. Diluted RSV F protein rabbit monoclonal antibodies labeled with horseradish peroxidase (1:1000, Sino Biological, Inc.) were added at 100 l/well and incubated at 37 C. for 1 h. The plate was washed by the PBST. Substrate solutions A and B were added in sequence for color development at room temperature for 5 min. A stop solution C was added to terminate chromogenic reaction. Optical density (OD) values were read at wavelengths of 450 nm and 630 nm.

    (2) Assay of His-Tag Antibody Binding Activity

    [0314] Cell secretion supernatant of the centrifuged recombinant expression mutants (including the wild-type RSV F protein 1 before artificial mutation) was 10-fold diluted by using the coating buffer and then coated a 96-well ELISA plate. 2 g/ml wild-type RSV F protein 2 before artificial mutation was used as a control well. Conditions were 100 l/well at 4 C. for 8 h to 12 h. The plate was washed by the PBST. The blocking buffer was added at 100 l/well and blocked at 37 C. for 2 h. The plate was washed by the PBST. Diluted His-tag antibodies (1:2000 dilution, Sino Biological, Inc.) were added at 100 l/well and incubated at 37 C. for 1 h. The plate was washed by the PBST. Diluted goat anti-mouse secondary antibodies labeled with horseradish peroxidase (1:10000, ZSBIO) were added at 100 l/well and incubated at 37 C. for 1 h. The plate was washed by the PBST. The substrate solutions A and B were added in sequence for color development at room temperature for 5 min. The stop solution C was added to terminate chromogenic reaction. Optical density (OD) values were read at wavelengths of 450 nm and 630 nm.

    [0315] ELISA results are shown in Table 6, Table 7 and Table 8. The results show that His-tag antibody assay of most mutants has high OD values, indicating that most of the constructed mutants can be effectively expressed. Table 6 shows that the wild-type RSV F protein 1 before artificial mutation and the wild-type RSV F protein 2 before artificial mutation have no AM14 monoclonal antibody binding activity. Table 7 and Table 8 show that a plurality of single-site mutants at the head domain and a plurality of multi-site combination mutants at the head domain can bind to the AM14 monoclonal antibodies. The mutants include mutants FHS-1, FHS-4, FHS-5, FHS-6, FHS-11, FHS-12, FHS-14, FHS-26, FHS-45, FHS-46, FHS-47, FHS-48, FHS-49, FHS-50, FHS-65, FHS-66, FHS-67, FHS-68, FHS-69, FHS-76, FHS-78, FHS-84, FHS-87, FHS-88, FHM-1, FHM-2, FHM-5, FHM-9, FHM-12, FHM-13, FHM-15, FHM-16, FHM-18, FHM-19, FHM-20, FHM-22, FHM-23, FHM-24 and FHM-25. The OD values of the AM14 monoclonal antibody assay of these single-site mutants or multi-site combination mutants were significantly higher than those of the wild-type RSV F protein 1 before artificial mutation and the wild-type RSV F protein 2 before artificial mutation, indicating that the RSV F protein is successfully stabilized in the prefusion trimer state by introducing the above mutation.

    TABLE-US-00006 TABLE 6 Binding activity of wild-type RSV F protein 1 before artificial mutation and wild-type RSV F protein 2 before artificial mutation with AM14 monoclonal antibody and His-tag antibody AM14 AM14 monoclonal His-tag monoclonal His-tag Protein antibody antibody Protein antibody antibody name (OD value) (OD value) name (OD value) (OD value) Wild-type 0.014 0.999 Wild-type 0.010 1.448 RSV F protein RSV F protein 1 before 2 before artificial artificial mutation mutation

    TABLE-US-00007 TABLE 7 Binding activity of illustrative mutant of single-site mutation introduced into head domain of RSV F protein with AM14 monoclonal antibody and His-tag antibody AM14 monoclonal His-tag Mutant antibody antibody number (OD value) (OD value) FHS-1 2.030 0.963 FHS-2 0.012 0.249 FHS-3 0.076 0.554 FHS-4 2.256 1.452 FHS-5 2.239 1.640 FHS-6 2.646 1.253 FHS-7 0.011 0.364 FHS-8 0.013 0.343 FHS-9 0.009 0.079 FHS-10 0.069 0.228 FHS-11 2.043 1.009 FHS-12 1.238 1.208 FHS-13 0.013 0.161 FHS-14 2.612 0.968 FHS-15 0.091 0.351 FHS-16 0.009 0.112 FHS-17 0.009 0.137 FHS-18 0.009 0.109 FHS-19 0.007 0.187 FHS-20 0.009 0.195 FHS-21 0.008 0.121 FHS-22 0.010 0.115 FHS-23 0.008 0.326 FHS-24 0.010 0.503 FHS-25 0.007 0.109 FHS-26 0.841 1.751 FHS-27 0.005 0.097 FHS-28 0.008 0.107 FHS-29 0.007 0.099 FHS-30 0.009 0.095 FHS-31 0.007 0.102 FHS-32 0.008 0.099 FHS-33 0.005 0.071 FHS-34 0.007 0.103 FHS-35 0.007 0.103 FHS-36 0.009 0.123 FHS-37 0.007 0.111 FHS-38 0.008 0.097 FHS-39 0.007 0.365 FHS-40 0.009 0.571 FHS-41 0.006 0.368 FHS-42 0.010 0.185 FHS-43 0.006 0.349 FHS-44 0.039 0.494 FHS-45 0.120 1.135 FHS-46 2.410 1.630 FHS-47 1.317 1.462 FHS-48 2.422 0.732 FHS-49 1.957 0.721 FHS-50 2.296 1.013 FHS-51 0.006 0.353 FHS-52 0.007 0.291 FHS-53 0.008 0.348 FHS-54 0.012 0.111 FHS-55 0.009 2.217 FHS-56 0.009 0.117 FHS-57 0.007 0.118 FHS-58 0.011 0.372 FHS-59 0.006 0.804 FHS-60 0.007 0.229 FHS-61 0.008 0.721 FHS-62 0.010 0.136 FHS-63 0.006 0.139 FHS-64 0.008 1.194 FHS-65 1.732 0.789 FHS-66 1.750 1.093 FHS-67 1.859 0.537 FHS-68 1.989 0.937 FHS-69 1.875 0.750 FHS-70 0.007 0.233 FHS-71 0.012 0.489 FHS-72 0.007 0.132 FHS-73 0.016 0.181 FHS-74 0.023 1.149 FHS-75 0.028 0.778 FHS-76 0.821 0.322 FHS-77 0.060 0.687 FHS-78 1.235 1.200 FHS-79 0.008 0.500 FHS-80 0.005 0.504 FHS-81 0.008 0.349 FHS-82 0.063 0.958 FHS-83 0.064 1.740 FHS-84 1.006 1.549 FHS-85 0.008 0.217 FHS-86 0.062 1.172 FHS-87 0.239 1.536 FHS-88 2.296 0.623 FHS-89 0.006 0.675 FHS-90 0.007 2.138 FHS-91 0.008 0.810 FHS-92 0.012 1.159 FHS-93 0.009 0.422 FHS-94 0.009 0.714 FHS-95 0.007 1.721 FHS-96 0.011 0.866 FHS-97 0.006 0.878 FHS-98 0.007 0.288

    TABLE-US-00008 TABLE 8 Binding activity of illustrative mutant of multi-site combination mutation introduced into head domain of RSV F protein with AM14 monoclonal antibody and His-tag antibody AM14 monoclonal His-tag antibody antibody Mutant number (OD value) (OD value) FHM-1 2.288 0.489 FHM-2 2.228 1.585 FHM-3 0.070 0.359 FHM-4 0.028 0.183 FHM-5 1.964 0.982 FHM-6 0.005 0.098 FHM-7 0.007 0.196 FHM-8 0.016 0.465 FHM-9 2.121 1.446 FHM-10 0.010 0.136 FHM-11 0.006 0.139 FHM-12 2.128 1.708 FHM-13 1.070 0.443 FHM-14 0.008 0.458 FHM-15 1.505 1.720 FHM-16 0.260 1.280 FHM-17 0.006 0.654 FHM-18 1.797 0.870 FHM-19 0.367 0.228 FHM-20 0.607 0.586 FHM-21 0.007 0.159 FHM-22 1.978 1.765 FHM-23 1.887 1.620 FHM-24 1.610 0.932 FHM-25 2.419 1.728

    Example 2 Purification, Identification and Biological Activity Evaluation of Illustrative Mutants FHM-24 and FHM-25 with Mutation Introduced into the Head Domain Expressed in Example 1

    [0316] Recombinant expression plasmids of constructed illustrative mutants FHM-24 and FHM-25 (corresponding amino acid sequences SEQ ID No. 156 and SEQ ID No. 157) with the mutation introduced into the head domain of the RSV F protein were transiently expressed by the Expi293 expression system (Thermo Fisher). Secretion supernatant was collected on the 5th day after transfection. Affinity chromatography purification was performed on the transfection supernatant by using a nickel column (HisTrap FF crude pre-packed column). An equilibration buffer (20 mM phosphate-150 mM sodium chloride, pH 7.4) was used for equilibrating the chromatographic column. A sample was loaded. A target protein was collected after elution in 20% and 50% elution buffers (20 mM phosphate-150 mM sodium chloride-500 mM imidazole, pH 7.4). The purified protein was subjected to 12% SDS-PAGE analysis. Results were shown in FIG. 3. A molecular weight corresponding to a target protein monomer band was about 50 kD to 70 kD, which was basically consistent with a theoretical value, indicating that the mutants FHM-24 and FHM-25 can be expressed correctly.

    [0317] After purified samples of the mutants FHM-24 and FHM-25 were obtained, the binding activity of the two mutants with the AM14 monoclonal antibody was detected by antibody binding experiments by using prefusion trimer specific-monoclonal antibody AM14 to verify whether the mutants were successfully stabilized in prefusion trimer state. Moreover, folding of the mutants FHM-24 and FHM-25 was further verified by using Synagis. Synagis can bind specifically to epitope II of the RSV F protein. The prefusion F protein and the postfusion F protein contain epitope II in a same conformation. Thus Synagis can bind to the prefusion F protein and the postfusion F protein. In the experiment, the wild-type RSV F protein 1 before artificial mutation and the wild-type RSV F protein 2 before artificial mutation were used as controls. A specific operation process of the experiment is as follows: the purified mutants FHM-24 and FHM-25, the wild-type RSV F protein 1 before artificial mutation, and the wild-type RSV F protein 2 before artificial mutation were subjected to 2-fold serial dilutions from initial concentrations of 80 g/ml (corresponding primary antibody was AM14 monoclonal antibody) and 10 g/ml (corresponding primary antibody was Synagis) by using a coating buffer and coated an ELISA plate at 100 l/well. 3 duplicate wells were made for each dilution. Coating was performed for 8 h to 12 h at 4 C. Blank wells were use as a negative control. The plate was washed by a PBST. A blocking buffer was added and blocked at 37 C. for 2 h. The plate was washed by the PBST. Diluted AM14 monoclonal antibodies (0.1 ug/ml) or Synagis (2 ug/ml) was added at 100 l/well and incubated at 37 C. for 1 h. The plate was washed by the PBST. Diluted goat anti-human secondary antibody labeled with horseradish peroxidase (1:10000) was added at 100 l/well and incubated at 37 C. for 1 h. The plate was washed by the PBST. Substrate solutions A and B were added in sequence for color development at room temperature for 5 min. A stop solution C was added to terminate chromogenic reaction. OD values were read at wavelengths of 450 nm and 630 nm. Mean OD.sub.450/630 nm values of the duplicate wells with different concentrations were calculated, and a standard deviation was calculated. With protein concentrations as an abscissa and OD.sub.450/630 nm mean values corresponding to concentrations as an ordinate, a broken line graph was drawn by Origin software.

    [0318] Results of antibody binding experiments were shown in FIG. 4 and FIG. 5. The results of the AM14 monoclonal antibody binding experiment (FIG. 4) show that the wild-type RSV F protein 1 before artificial mutation and the wild-type RSV F protein 2 before artificial mutation have no AM14 monoclonal antibody binding activity, and the illustrative mutants FHM-24 and FHM-25 with the mutation introduced into the head domain exhibit AM14 monoclonal antibody binding activity. The results indicate that the RSV F protein can be stabilized in the prefusion trimer state by introducing the stabilizing mutation into the head domain, to prevent the transition conformational from a prefusion conformation to a postfusion conformation. The results of the Synagis binding experiment (FIG. 5) show that the mutants FHM-24 and FHM-25, the wild-type RSV F protein 1 before artificial mutation and the wild-type RSV F protein 2 before artificial mutation all have Synagis binding activity. All the four proteins can fold correctly.

    Example 3: On the Basis of Introducing Mutation into a Head Domain of an RSV F Protein, Mutation Design of a Tail Domain is Added to Further Enhance Stability of a Prefusion RSV F Protein Trimer, and Further to Improve Immunogenicity

    [0319] Stability of a prefusion RSV F protein trimer structure has an important influence on the immunogenicity of the trimer as an antigen. On the basis of the mutation introduced into the head domain in the above examples, mutation was also introduced in a tail domain of the F protein in the example. By enhancing an interaction between triple helix bundles in RR2, the stability of the F protein trimer structure was improved, and AM14 monoclonal antibody binding activity was enhanced, so as to further improve the immunogenicity. It was reported that the AM14 monoclonal antibody binding activity positively correlated with a neutralizing antibody titer produced by immunized animals (Ref: Che Y. et al. Sci. Transl. Med., 2023, 15:eade6422.). Thus, in the example, single-site mutation or a multi-site mutation combination was introduced into RR2 of the tail domain of the RSV F protein to further enhance the stability of the prefusion F protein trimer structure by enhancing the interaction between the triple helix bundles in RR2. In this example, mutants FHM-24 (mutation of 227N.fwdarw.M and 230L.fwdarw.F was introduced in the head domain) and FHM-25 (mutation of 227N.fwdarw.M, 230L.fwdarw.F, 190S.fwdarw.V and 193L.fwdarw.M was introduced into the head domain) of mutation introduced into the head domain were taken as instances to further enhance the stability of the RSV F protein trimer by further introducing the stability mutation into the tail domain. Some illustrative single-site mutation design of the tail domain is shown in Table 9. The single-site mutation of the tail domain can be combined arbitrarily and can have similar or stronger stabilizing effects. Some illustrative mutation combinations are shown in Table 10 and Table 11.

    TABLE-US-00009 TABLE 9 Illustrative single-site mutation design introduced into the tail domain based on the mutant FHM-24, to further enhance the stability of the prefusion RSV F protein trimer Mutation Single-site mutation Mutant introduced into introduced into number the head domain the tail domain FHCS-1 227N.fwdarw.M, 230L.fwdarw.F 509S.fwdarw.M FHCS-2 227N.fwdarw.M, 230L.fwdarw.F 509S.fwdarw.F FHCS-3 227N.fwdarw.M, 230L.fwdarw.F 509S.fwdarw.L FHCS-4 227N.fwdarw.M, 230L.fwdarw.F 505F.fwdarw.W FHCS-5 227N.fwdarw.M, 230L.fwdarw.F 502S.fwdarw.Y

    TABLE-US-00010 TABLE 10 Illustrative site mutation combination design introduced into the tail domain based on the mutant FHM-24, to further enhance the stability of the prefusion RSV F protein trimer Mutation Site mutation combination Mutant introduced into introduced into number the head domain the tail domain FHCM-1 227N.fwdarw.M, 230L.fwdarw.F 505F.fwdarw.W, 509S.fwdarw.L FHCM-2 227N.fwdarw.M, 230L.fwdarw.F 501Q.fwdarw.M, 509S.fwdarw.F

    TABLE-US-00011 TABLE 11 Illustrative site mutation design introduced into the tail domain based on the mutant FHM-25, to further enhance the stability of the prefusion RSV F protein trimer Mutation Site mutation combination Mutant introduced into introduced into number the head domain the tail domain FHCM-3 227N.fwdarw.M, 230L.fwdarw.F, 505F.fwdarw.W, 509S.fwdarw.L 190S.fwdarw.V, 193L.fwdarw.M FHCM-4 227N.fwdarw.M, 230L.fwdarw.F, 501Q.fwdarw.M, 509S.fwdarw.F 190S.fwdarw.V, 193L.fwdarw.M

    Example 4 Plasmid Construction, Recombinant Expression, Purification and Biological Activity Validation of the Mutant Designed in Example 3

    [0320] In order to verify whether the mutation introduced into the tail domain can further enhance the stability of the prefusion RSV F protein trimer, the example used a prefusion F protein trimer-specifically binding monoclonal antibody AM14 to assay antibody binding activity of the mutant designed in Example 3. The antibody binding activity was compared with AM14 antibody binding activity of the mutants FHM-24 or FHM-25 without mutation introduced into the tail domain, the wild-type RSV F protein 1 before artificial mutation and the wild-type RSV F protein 2 before artificial mutation to evaluate an enhancement effect of the mutation introduced into the tail domain on the stability of the prefusion RSV F protein trimer.

    [0321] First, amino acid sequences corresponding to the mutants of the RSV F protein designed in Example 3 were SEQ ID No. 158-166 in sequence. According to the corresponding amino acid sequences, the signal peptide suitable for the recombinant expression system HEK293, the T4 foldon trimerization motif, the linker, and an additional tag sequence useful for purification were added. Codon optimization was performed according to a codon bias of the HEK293 to form corresponding coding gene sequences. Recombinant expression plasmids were constructed by using the plasmid construction and recombinant expression processes described in Example 1. Recombinant expression of the mutants was performed by using an Expi293 transient expression system. Cell secretion supernatant was collected on the 5th day after transfection.

    [0322] Furthermore, all mutants designed in Example 3 were purified through a purification method described in Example 1, to obtain a purified mutant protein. The purified protein was subjected to 12% SDS-PAGE analysis. Results were shown in FIG. 6. A molecular weight corresponding to a target protein monomer band was about 50 kD to 70 kD, indicating that the mutants can be expressed correctly.

    [0323] Finally, AM14 monoclonal antibody binding activity was assayed for a purified mutant sample obtained by purification through the ELISA described in Example 2 by using a prefusion RSV F protein trimer-specifically binding monoclonal antibody AM14. The AM14 monoclonal antibody binding activity was compared with AM14 monoclonal antibody binding activity of the mutants FHM-24 or FHM-25 without mutation introduced into the tail domain, the wild-type RSV F protein 1 before artificial mutation, and the wild-type RSV F protein 2 before artificial mutation to evaluate the enhancement effect of the mutation introduced into the tail domain on the stability of the prefusion RSV F protein trimer. Moreover, folding of the constructed mutants was further verified by using Synagis.

    [0324] Results of an AM14 monoclonal antibody binding experiment were shown in FIG. 7A and FIG. 7B. The results show that the wild-type RSV F protein 1 before artificial mutation and the wild-type RSV F protein 2 before artificial mutation have no AM14 monoclonal antibody binding activity. The F protein mutants with mutation introduced into the head domain and the tail domain have significantly increased AM14 monoclonal antibody binding activity compared with the mutants (FHM-24, FHM-25) without mutation introduced into the tail domain, the wild-type RSV F protein 1 before artificial mutation, and the wild-type RSV F protein 2 before artificial mutation. The results indicate that the mutation introduced into the tail domain can further enhance the stability of the prefusion RSV F protein trimer structure. Results of a Synagis binding experiment were shown in FIG. 8A and FIG. 8B. The results show that the wild-type RSV F protein 1 before artificial mutation, the wild-type RSV F protein 2 before artificial mutation, the F protein mutants without mutation introduced into the tail domain, and the mutants with stability mutation introduced into the tail domain have strong Synagis binding activity, indicating that these proteins can fold correctly.

    Example 5 an Accelerated Stability Test Study of Illustrative Constructed Mutants of an RSV F Protein

    [0325] The stability of the constructed mutants of the RSV F protein is an important characteristic to be paid attention when the mutants are used in vaccine development as antigens. In the example, a thermal accelerated stability test study was carried out on constructed illustrative mutants FHCS-1, FHCS-2 and FHCS-4 of the RSV F protein. Mutant samples obtained after expression purification were stored at 37 C. for 2 weeks and 4 weeks respectively. Meanwhile, same samples were stored at 80 C. for 2 weeks and 4 weeks as controls. Stability samples were subjected to 2-fold serial dilutions from initial concentrations of 10 g/ml (corresponding primary antibody was AM14 monoclonal antibody) and 2.5 g/ml (corresponding primary antibody was Synagis) by using a coating buffer and coated an ELISA plate at 100 l/well. 2 duplicate wells were prepared for each dilution. AM14 monoclonal antibody binding activity and Synagis binding activity of the samples were assayed through the ELISA method described in Example 2.

    [0326] The results of a thermal accelerated stability test were shown in Table 12, Table 13, Table 14, and Table 15. The results show that after the mutants of the RSV F protein constructed in the present disclosure are placed at 37 C. for 4 weeks, the AM14 monoclonal antibody binding activity and the Synagis binding activity do not change obviously compared with the samples placed at 80 C., indicating that prefusion trimer structures of the mutants of the RSV F protein constructed in the present disclosure are still stable after being placed at 37 C. for 4 weeks.

    TABLE-US-00012 TABLE 12 Thermal accelerated stability result of illustrative mutant of RSV F protein at 3 antibody binding activity (OD Value)(per sample was assayed in duplicate) Protein concentration (ng/ml) 80 C. text missing or illegible when filed Solutionname 10000 5000 2500 1250 625 312.5 10000 500text missing or illegible when filed FHCS-1 1.821 0.158 0.021 0.006 0.003 0.004 2.278 0.27text missing or illegible when filed 2.117 0.167 0.030 0.006 0.005 0.008 2.472 0.24text missing or illegible when filed FHCS-2 1.878 0.203 0.020 0.009 0.005 0.005 2.142 0.35text missing or illegible when filed 2.147 0.382 0.024 0.013 0.006 0.005 1.887 0.35text missing or illegible when filed FHCS-4 2.128 0.404 0.040 0.013 0.007 0.004 2.290 0.81text missing or illegible when filed 1.929 0.294 0.028 0.011 0.007 0.007 2.206 0.74text missing or illegible when filed text missing or illegible when filed indicates data missing or illegible when filed

    TABLE-US-00013 TABLE 13 Thermal accelerated stability result of illustrative mutant of RSV F protein at 37 C. for 2 weeks-Synagis binding activity (OD Value)(per sample was assayed in duplicate) Protein concentration (ng/ml) 80 C. 37 C. 2 W Solution name 2500 1250 625 312.5 156.3 78.13 2500 1250 625 312.5 156.3 78.13 FHCS-1 1.355 0.357 0.170 0.340 0.027 0.009 1.882 0.478 0.123 0.051 0.017 0.010 1.347 0.350 0.181 0.043 0.008 0.010 1.430 0.493 0.125 0.050 0.018 0.015 FHCS-2 1.650 0.572 0.209 0.089 0.028 0.014 1.605 0.780 0.211 0.084 0.037 0.018 1.908 0.767 0.208 0.108 0.032 0.015 1.846 0.704 0.227 0.095 0.036 0.022 FHCS-4 0.825 0.252 0.094 0.052 0.021 0.009 1.461 0.535 0.187 0.074 0.033 0.018 0.743 0.209 0.066 0.033 0.017 0.011 1.581 0.540 0.166 0.071 0.024 0.015

    TABLE-US-00014 TABLE 14 Thermal accelerated stability result of illustrative mutant of RSV F protein at 37 C. for 4 weeks-AM14 monoclonal antibody binding activity (OD Value)(per sample was assayed in duplicate) Protein concentration (ng/ml) 80 C. 37 C. 4 W Solutionname 10000 5000 2500 1250 625 312.5 10000 5000 2500 1250 625 312.5 FHCS-1 1.775 0.902 0.034 0.011 0.004 0.003 1.872 1.166 0.109 0.018 0.006 0.005 2.066 0.775 0.039 0.015 0.007 0.006 2.167 1.332 0.096 0.024 0.008 0.005 FHCS-2 2.012 0.985 0.051 0.017 0.006 0.005 2.110 1.949 0.450 0.025 0.005 0.005 2.112 0.796 0.056 0.020 0.008 0.007 2.225 1.480 0.185 0.019 0.006 0.005 FHCS-4 0.825 0.252 0.008 0.007 0.005 0.008 1.761 0.665 0.045 0.007 0.006 0.004 0.743 0.209 0.007 0.006 0.006 0.006 2.009 0.438 0.019 0.008 0.007 0.007

    TABLE-US-00015 TABLE 15 Thermal accelerated stability result of illustrative mutant of RSV F protein at 37 C. for 4 weeks-Synagis binding activity (OD Value)(per sample was assayed in duplicate) Protein concentration (ng/ml) 80 C. 37 C. 4 W Solution name 2500 1250 625 312.5 156.3 78.13 2500 1250 625 312.5 156.3 78.13 FHCS-1 1.504 0.635 0.161 0.065 0.021 0.010 1.909 1.168 0.324 0.142 0.055 0.025 1.612 0.642 0.154 0.053 0.019 0.010 2.109 1.221 0.612 0.168 0.062 0.032 FHCS-2 1.783 0.767 0.409 0.109 0.043 0.016 1.823 0.995 0.420 0.154 0.069 0.030 1.799 0.928 0.307 0.097 0.038 0.016 1.837 0.910 0.400 0.143 0.058 0.028 FHCS-4 0.947 0.321 0.094 0.032 0.017 0.009 1.803 1.048 0.378 0.138 0.050 0.019 0.991 0.328 0.110 0.028 0.016 0.008 1.777 0.985 0.389 0.120 0.048 0.025

    Example 6 Evaluation of Animal Immunization Effects of Illustrative Constructed Mutants of an RSV F Protein

    [0327] In order to verify whether the immunization effects of the mutants of the RSV F protein constructed in the present disclosure is significantly enhanced, animal immunization effect evaluation studies were carried out on mutants FHCS-1, FHCS-2, FHCS-3 and FHCS-4 in the example. The immunization effects were compared with immunization effects of a wild-type RSV F protein before artificial mutation and a mutant protein FHM-24 with mutation only introduced into a head domain.

    [0328] A specific experimental method was as follows: a purified illustrative mutant of the F protein and the wild-type RSV F protein 2 before artificial mutation were mixed with an aluminum hydroxide adjuvant separately, and then a polyinosinic solution was added to prepare vaccines. BALB/c mice (purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., SPF grade, female, 6 weeks to 8 weeks old) were immunized with the prepared vaccines, and each group had 10 mice. The mice were immunized intramuscularly at week 0 and week 2 with 0.5 ug/mouse antigen and 0.1 ml/mouse immune volume. Blood samples were collected 1 week after the last immunization. A live RSV strain A2 neutralization experiment was used to assay a neutralizing antibody level in immune serum. Specifically, a maintenance medium (DMEM medium containing 2% fetal bovine serum and 1% streptomycin) was used for 2-fold serial dilutions on the serum. Diluted RSV A2 viruses were added at 100 TCID.sub.50/well. Duplicate wells were made for each piece of serum. A virus control well without serum and a cell control well without serum and viruses were set. Incubation was performed at 4 C. for 2 h. A Hep2 cell suspension was added at 2*104 cells per well and cultured in 37 C., in a 5% CO2 incubator for 3 days. Neutralization results were determined through an immunofluorescence method. A serum dilution multiple lower than 50% of a mean value of fluorescent spots in the virus control well was taken as a neutralizing antibody titer of the serum.

    [0329] Results were shown in FIG. 9, compared with the wild-type RSV F protein before artificial mutation, the mutant had an increased neutralizing antibody titer after mutation was introduced into the head domain. A geometric mean titer (GMT) of neutralizing antibodies increased from 147 to 239. Furthermore, after the mutation was introduced into the head domain and the tail domain, the neutralizing antibody titer was further increased significantly. The GMT increased to 588 to 1261, which was 4 times to 8.58 times higher than that of the wild-type RSV F protein before artificial mutation. The results show that the RSV F protein is stabilized in the prefusion trimer structure state by introducing the mutation into the head domain and the tail domain. Immunogenicity of the protein is obviously improved. Candidate antigens with obvious technical advantages and immunoactivity advantages are provided for development of recombinant RSV vaccines.

    [0330] What are described above are merely preferred embodiments of the present disclosure. It should be pointed out that those of ordinary skill in the art can make several improvements and modifications without departing from the principles of the present disclosure, and these improvements and modifications should also be deemed as falling within the scope of protection of the present disclosure.