Mutant of L1 protein of human papillomavirus type 18

Abstract

The present invention relates to a mutated HPV18 L1 protein (or a variant thereof), a sequence encoding the same, a method for preparing the same, and a virus-like particle comprising the same, wherein the protein (or a variant thereof) and the virus-like particle can induce the generation of neutralizing antibodies against at least two HPV types (for example, HPV18 and HPV45, or HPV18, HPV45 and HPV59), and therefore can be used to prevent infection by said at least two HPV types, and a disease caused by said infection, such as cervical cancer and condyloma acuminatum. The invention further relates to the use of the protein and the virus-like particle in the manufacture of a pharmaceutical composition or a vaccine for preventing infection by said at least two HPV types, and a disease caused by said infection, such as cervical cancer and condyloma acuminatum.

Claims

1. A mutated HPV18 L1 protein, wherein as compared with a wild type HPV18 L1 protein, (I) the mutated HPV18 L1 protein has the following mutations: (1) N-terminal truncation of 40-80 amino acids; and (2) (a) substitution of amino acid residues at positions of the wild type HPV18 L1 protein which correspond to positions 235-243 of SEQ ID NO: 1 with amino acid residues at the corresponding positions of a L1 protein of a second type of wild-type HPV; or (b) substitution of amino acid residues at positions of the wild type HPV18 L1 protein which correspond to positions 327-346 of SEQ ID NO: 1 with amino acid residues at the corresponding positions of a L1 protein of a second type of wild-type HPV; or (c) substitution of amino acid residues at positions of the wild type HPV18 L1 protein which correspond to positions 114-123 of SEQ ID NO: 1 with amino acid residues at the corresponding positions of a L1 protein of a second type of wild-type HPV; or (d) substitution of amino acid residues at positions of the wild type HPV18 L1 protein which correspond to positions 176-202 of SEQ ID NO: 1 with amino acid residues at the corresponding positions of a L1 protein of a second type of wild-type HPV, or, (II) the mutated HPV18 L1 protein has the mutations as defined in (1) and (2) (a), and further has the following mutation: (3) substitution of amino acid residues at positions of the wild type HPV18 L1 protein which correspond to positions 112-123 of SEQ ID NO: 1 with amino acid residues at the corresponding positions of a L1 protein of a third type of wild-type HPV; or, (III) the mutated HPV18 L1 protein has the mutations as defined in (1) and (2) (b), and further has the following mutation: (3) substitution of amino acid residues at positions of the wild type HPV 18 L1 protein which correspond to positions 112-123 of SEQ ID NO: 1 with amino acid residues at the corresponding positions of a L1 protein of a third type of wild-type HPV; or, (IV) the mutated HPV18 L1 protein has the mutations as defined in (1) and (2) (c), and further has the following mutation: (4) substitution of amino acid residues at positions of the wild type HPV18 L1 protein which correspond to positions 410-421 of SEQ ID NO: 1 with amino acid residues at the corresponding positions of a L1 protein of a third type of wild-type HPV; or, (V) the mutated HPV18 L1 protein has the mutations as defined in (1) and (2) (d), and further has the mutation as defined in (4); or, (VI) the mutated HPV18 L1 protein has the mutations as defined in (1), (2) (c) and (2) (a), and optionally, further has the mutation as defined in (4); wherein said corresponding positions are determined by optimal alignment of the sequences being compared.

2. An isolated nucleic acid, encoding the mutated HPV18 L1 protein according to claim 1.

3. A vector comprising the isolated nucleic acid according to claim 2.

4. A host cell comprising the isolated nucleic acid according to claim 2 and/or a vector comprising the isolated nucleic acid according to claim 2.

5. An HPV virus-like particle, comprising or consisting of the mutated HPV18 L1 protein according to claim 1.

6. A composition, comprising: the mutated HPV18 L1 protein according to claim 1, or (ii) an isolated nucleic acid encoding the mutated HPV18 L1 protein as described in (i), or (iii) a vector comprising the isolated nucleic acid as described in (ii), or (iv) a host cell comprising the isolated nucleic acid as described in (ii) and/or the vector comprising the isolated nucleic acid as described in (iii), or (v) a HPV virus-like particle comprising or consisting of the mutated HPV18 L1 protein as described in (i).

7. A pharmaceutical composition or vaccine, comprising the HPV virus-like particle according to claim 5, and optionally a pharmaceutically acceptable carrier and/or excipient.

8. A method for preparing the mutated HPV18 L1 protein according to claim 1, comprising expressing the mutated HPV18 L1 protein in a host cell, and then recovering the mutated HPV18 L1 protein from a culture of the host cell.

9. A method for preparing a vaccine, comprising combining the HPV virus-like particle according to claim 5 with a pharmaceutically acceptable carrier and/or excipient.

10. A method for preventing HPV infection or a disease caused by HPV infection, comprising administering to a subject a prophylactically effective amount of the HPV virus-like particle according to claim 5 or a pharmaceutical composition or vaccine comprising the HPV virus-like particle according to claim 5 and optionally a pharmaceutically acceptable carrier and/or excipient.

11. The mutated HPV18 L1 protein according to claim 1, wherein the mutated HPV18 L1 protein is characterized by one or more of the following items: (i) the mutated HPV18 L1 protein has 45, 50, 52, 55, 58, 60, 62, 65, 68, 70, 72, 75 or 78 amino acids truncated at N-terminal, as compared with the wild type HPV18 L1 protein; (ii) the second type of wild-type HPV is HPV45; (iii) the amino acid residues at the corresponding positions as described in (2) (a) are amino acid residues at positions 201-209 of a wild type HPV45 L1 protein; (iv) the amino acid residues at the corresponding positions as described in (2) (b) are amino acid residues at positions 293-314 of a wild type HPV45 L1 protein; (v) the amino acid residues at the corresponding positions as described in (2) (c) are amino acid residues at positions 79-89 of a wild type HPV45 L1 protein; (vi) the amino acid residues at the corresponding positions as described in (2) (d) are amino acid residues at positions 142-168 of a wild type HPV45 L1 protein; (vii) the third type of wild-type HPV is HPV59; (viii) the amino acid residues at the corresponding positions as described in (3) are amino acid residues at positions 51-62 of a wild type HPV59 L1 protein; (ix) the amino acid residues at the corresponding positions as described in (4) are amino acid residues at positions 349-360 of a wild type HPV59 L1 protein; (x) the wild type HPV18 L1 protein has an amino acid sequence as set forth in SEQ ID NO: 1; (xi) the wild type HPV45 L1 protein has an amino acid sequence as set forth in SEQ ID NO: 2; (xii) the wild type HPV59 L1 protein has an amino acid sequence as set forth in SEQ ID NO: 3.

12. The mutated HPV18 L1 protein according to claim 1, wherein the mutated HPV18 L1 protein has an amino acid sequence selected from the group consisting of: SEQ ID NO: 6, 7, 9, 13, 17, 18 and 19.

13. The isolated nucleic acid according to claim 2, wherein the isolated nucleic acid has a nucleotide sequence selected from the group consisting of: SEQ ID NO: 25, 26, 28, 32, 36, 37 and 38.

14. The pharmaceutical composition or vaccine according to claim 7, wherein the HPV virus-like particle is present in an amount effective for preventing HPV infection or a disease caused by HPV infection.

15. The pharmaceutical composition or vaccine according to claim 14, wherein the HPV infection is infection by one or more HPV types, and/or, the disease caused by HPV infection is selected from the group consisting of cervical cancer and condyloma acuminatum.

16. The pharmaceutical composition or vaccine according to claim 15, wherein the HPV infection is selected from: HPV18 infection, HPV45 infection, HPV59 infection and any combination thereof.

17. The method according to claim 8, wherein the host cell is E. coli.

18. The method according to claim 17, wherein the method comprises the steps of: expressing the mutated HPV18 L1 protein in E. coli, and then obtaining the mutated HPV18 L1 protein by purifying a lysate supernatant of the E. coli.

19. The method according to claim 10, wherein the HPV infection is infection by one or more HPV types, and/or, the disease caused by HPV infection is selected from the group consisting of cervical cancer and condyloma acuminatum.

20. The method according to claim 19, wherein the HPV infection is selected from: HPV18 infection, HPV45 infection, HPV59 infection and any combination thereof.

Description

DESCRIPTION OF DRAWINGS

(1) FIG. 1 shows the SDS-PAGE result of the purified mutated proteins in Example 1. Lane M: protein molecular weight marker; Lane 1: HPV18N65 (HPV18 L1 protein having 65 amino acids truncated at N-terminal); Lane 2: H18N65-45T1; Lane 3: H18N65-45T2; Lane 4: H18N65-45T3; Lane 5: H18N65-45T4; Lane 6: H18N65-45T5; Lane 7: HPV18N65; Lane 8: H18N65-45T3-5951; Lane 9: H18N65-45T3-5952; Lane 10: H18N65-45T3-5954; Lane 11: H18N65-45T3-5955; Lane 12: H18N65-45T4-5951; Lane 13: H18N65-45T4-5952; Lane 14: H18N65-45T4-5953; Lane 15: HPV18N65; Lane 16: H18N65-45T4-5955; Lane 17: HPV18N65; Lane 18: H18N65-45T1-5955; Lane 19: H18N65-45T2-5955; Lane 20: H18N65-45T1T3-5955. The result showed that after chromatographic purification, H18N65-45T1, H18N65-45T2, H18N65-45T3, H18N65-45T4, H18N65-45T5, H18N65-45T3-5951, H18N65-45T3-5952, H18N65-45T3-5954, H18N65-45T3-5955, H18N65-45T4-5951, H18N65-45T4-5952, H18N65-45T4-5953, H18N65-45T4-5955, H18N65-45T1-5955, H18N65-45T2-5955 and H18N65-45T1T3-5955 protein reached a purity of above 85%.

(2) FIG. 2 shows the Western Blot result of H18N65-45T1, H18N65-45T2, H18N65-45T3, H18N65-45T4, H18N65-45T5, H18N65-45T3-5951, H18N65-45T3-5952, H18N65-45T3-5954, H18N65-45T3-5955, H18N65-45T4-5951, H18N65-45T4-5952, H18N65-45T4-5953, H18N65-45T4-5955, H18N65-45T1-5955, H18N65-45T2-5955 and H18N65-45T1T3-5955 prepared in Example 1, as determined by using a broad-spectrum antibody 4B3. Lane M: protein molecular weight marker; Lane 1: HPV18N65; Lane 2: H18N65-45T1; Lane 3: H18N65-45T2; Lane 4: H18N65-45T3; Lane 5: H18N65-45T4; Lane 6: H18N65-45T5; Lane 7: HPV18N65; Lane 8: H18N65-45T3-59S1; Lane 9: H18N65-45T3-59S2; Lane 10: H18N65-45T3-59S4; Lane 11: H18N65-45T3-59S5; Lane 12: H18N65-45T4-59S1; Lane 13: H18N65-45T4-59S2; Lane 14: H18N65-45T4-59S3; Lane 15: HPV18N65; Lane 16: H18N65-45T4-59S5; Lane 17: HPV18N65; Lane 18: H18N65-45T1-59S5; Lane 19: H18N65-45T2-59S5; Lane 20: H18N65-45T1T3-59S5. The result showed that the mutated proteins H18N65-45T1, H18N65-45T2, H18N65-45T3, H18N65-45T4, H18N65-45T5, H18N65-45T3-59S1, H18N65-45T3-59S2, H18N65-45T3-59S4, H18N65-45T3-59S5, H18N65-45T4-59S1, H18N65-45T4-59S2, H18N65-45T4-59S3, H18N65-45T4-59S5, H18N65-45T1-59S5, H18N65-45T2-59S5 and H18N65-45T1T3-59S5 could be specifically recognized by the broad-spectrum antibody 4B3.

(3) FIGS. 3A-3S show the results of the samples comprising the protein HPV18N65 (FIG. 3A), HPV45N27 (FIG. 3B), HPV59 L1 (FIG. 3C), H18N65-45T1 (FIG. 3D), H18N65-45T2 (FIG. 3E), H18N65-45T3 (FIG. 3F), H18N65-45T4 (FIG. 3G), H18N65-45T5 (FIG. 3H), H18N65-45T3-59S1 (FIG. 3I), H18N65-45T3-59S2 (FIG. 3J), H18N65-45T3-59S4 (FIG. 3K), H18N65-45T3-59S5 (FIG. 3L), H18N65-45T4-59S1 (FIG. 3M), H18N65-45T4-59S2 (FIG. 3N), H18N65-45T4-59S3 (FIG. 3O), H18N65-45T4-59S5 (FIG. 3P), H18N65-45T1-59S5 (FIG. 3Q), H18N65-45T2-59S5 (FIG. 3R), and H18N65-45T1T3-59S5 (FIG. 3S), as analyzed by molecular sieve chromatography. The results showed that the first protein peak of the samples comprising the protein H18N65-45T1, H18N65-45T2, H18N65-45T3, H18N65-45T4, H18N65-45T5, H18N65-45T3-59S1, H18N65-45T3-59S2, H18N65-45T3-59S4, H18N65-45T3-59S5, H18N65-45T4-59S1, H18N65-45T4-59S2, H18N65-45T4-59S3, H18N65-45T4-59S5, H18N65-45T1-59S5, H18N65-45T2-59S5, or H18N65-45T1T3-59S5 appeared at about 13 min, which was comparable to that of VLP assembled by HPV18N65 L1 protein (HPV18N65 VLP), VLP assembled by HPV45N27 protein (HPV45N27 VLP) and VLP assembled by HPV59 L1 protein (HPV59 VLP). This showed that all these proteins were able to assemble into VLPs.

(4) FIGS. 4A-4S show the transmission electron microscopy (TEM) photographs (taken at 100,000× magnification, Bar=0.1 μm) of various VLP samples. FIG. 4A, VLP assembled by HPV18N65; FIG. 4B, VLP assembled by HPV45N27; FIG. 4C, VLP assembled by HPV59 L1; FIG. 4D, VLP assembled by H18N65-45T1; FIG. 4E, VLP assembled by H18N65-45T2; FIG. 4F, VLP assembled by H18N65-45T3; FIG. 4G, VLP assembled by H18N65-45T4; FIG. 4H, VLP assembled by H18N65-45T5; FIG. 4I, VLP assembled by H18N65-45T3-59S1; FIG. 4J, VLP assembled by H18N65-45T3-59S2; FIG. 4K, VLP assembled by H18N65-45T3-59S4; FIG. 4L, VLP assembled by H18N65-45T3-59S5; FIG. 4M, VLP assembled by H18N65-45T4-59S1; FIG. 4N, VLP assembled by H18N65-45T4-59S2; FIG. 4O, VLP assembled by H18N65-45T4-59S3; FIG. 4P, VLP assembled by H18N65-45T4-59S5; FIG. 4Q, VLP assembled by H18N65-45T1-59S5; FIG. 4R, VLP assembled by H18N65-45T2-59S5; FIG. 4S, VLP assembled by H18N65-45T1T3-59S5. The results showed that H18N65-45T1, H18N65-45T2, H18N65-45T3, H18N65-45T4, H18N65-45T5, H18N65-45T3-59S1, H18N65-45T3-59S2, H18N65-45T3-59S4, H18N65-45T3-59S5, H18N65-45T4-59S1, H18N65-45T4-59S2, H18N65-45T4-59S3, H18N65-45T4-59S5, H18N65-45T1-59S5, H18N65-45T2-59S5 and H18N65-45T1T3-59S5 were similar to HPV18N65, HPV45N27 and HPV59 L1, and were able to assemble into VLPs with a radius of about 30 nm.

(5) FIGS. 5A-5H show the results of sedimentation velocity analysis of HPV18N65 VLP, HPV45N27 VLP, HPV59 VLP, H18N65-45T3 VLP, H18N65-45T4 VLP, H18N65-45T3-59S1 VLP, H18N65-45T4-59S1 VLP and H18N65-45T1T3-59S5 VLP. FIG. 5A, HPV18N65 VLP; FIG. 5B, HPV45N27 VLP; FIG. 5C, HPV59 VLP; FIG. 5D, H18N65-45T3 VLP; FIG. 5E, H18N65-45T4 VLP; FIG. 5F, H18N65-45T3-59S1 VLP; FIG. 5G, H18N65-45T4-59S1 VLP; FIG. 5H, H18N65-45T1T3-59S5 VLP. The results showed that the sedimentation coefficients of H18N65-45T3 VLP, H18N65-45T4 VLP and H18N65-45T1T3-59S5 VLP were 143.7S, 173.3S and 167.1S, respectively, which was similar to that of HPV18N65 VLP, HPV45N27 VLP and HPV59 VLP (HPV18N65 VLP, 142.2S; HPV45N27 VLP, 146.5S, and HPV59 VLP, 139.3S). This showed that H18N65-45T3, H18N65-45T4 and H18N65-45T1T3-59S5 were able to assemble into virus-like particles that were similar to wild type VLP in terms of size and morphology.

(6) FIGS. 6A-6H show the detection results of thermostability of HPV18N65 VLP, HPV45N27 VLP, HPV59 VLP, H18N65-45T3 VLP, H18N65-45T4 VLP, H18N65-45T3-59S1 VLP, H18N65-45T4-59S1 VLP, and H18N65-45T1T3-59S5 VLP. FIG. 6A, HPV18N65 VLP; FIG. 6B, HPV45N27 VLP; FIG. 6C, HPV59 VLP; FIG. 6D, H18N65-45T3 VLP; FIG. 6E, H18N65-45T4 VLP; FIG. 6F, H18N65-45T3-59S1 VLP; FIG. 6G, H18N65-45T4-59S1 VLP; FIG. 6H, H18N65-45T1T3-59S5 VLP. The results showed that all the VLPs formed by these proteins had very high thermostability.

(7) FIG. 7A shows the evaluation result of immune protection of H18N65-45T1 VLP, H18N65-45T2 VLP, H18N65-45T3 VLP, H18N65-45T4 VLP and H18N65-45T5 VLP in mice of the Experimental groups, and of HPV18N65 VLP, HPV45N27 VLP, and the mixed HPV18/HPV45 VLP in mice of the Control groups. The result showed that H18N65-45T1 VLP, H18N65-45T2, VLPH18N65-45T3 VLP and H18N65-45T4 VLP each retained the activity that could induce the generation of high-titer neutralizing antibodies against HPV18 in mice, and their protective effects against HPV18 were comparable to that of HPV18N65 VLP alone and that of the mixed HPV18/HPV45 VLP, and their activities that could induce the generation of neutralizing antibodies against HPV45 in mice were higher than that of HPV18N65 VLP alone. In particular, H18N65-45T3 VLP and H18N65-45T4 VLP each could induce the generation of high-titer neutralizing antibodies against HPV45 and HPV18 in mice; and their protective effects against HPV18 were comparable to that of HPV18N65 VLP alone and that of the mixed HPV18/HPV45 VLP, and were significantly higher than that of HPV45N27 VLP alone; and their protective effects against HPV45 were comparable to that of HPV45N27 VLP alone and that of the mixed HPV18/HPV45 VLP, and were significantly higher than that of HPV18N65 VLP alone. This showed that after mutation, H18N65-45T1 VLP, H18N65-45T2, VLPH18N65-45T3 VLP and H18N65-45T4 VLP retained their immunogenicity against HPV18, and their immunogenicity against HPV45 is also improved as compared to HPV18N65 VLP. Especially, H18N65-45T3 VLP and H18N65-45T4 VLP had good cross-immunogenicity and cross-protection against HPV18 and HPV45, and could be used as effective vaccines for preventing HPV18 infection and/or HPV45 infection, and could be used in place of a mixed vaccine comprising HPV18 VLP and HPV45 VLP.

(8) FIG. 7B shows the evaluation result of immune protection of H18N65-45T3-59S1 VLP, H18N65-45T3-59S2 VLP, H18N65-45T3-59S4 VLP, H18N65-45T3-59S5 VLP, H18N65-45T4-59S1 VLP, H18N65-45T4-59S2 VLP, H18N65-45T4-59S3 VLP and H18N65-45T4-59S5 VLP in mice of the Experimental groups, and of HPV18N65 VLP, HPV45N27 VLP, HPV59 VLP and the mixed HPV18/HPV45/HPV59 VLP in mice of the Control groups. The result showed that H18N65-45T3-59S1 VLP and H18N65-45T4-59S1 VLP each could induce the generation of high-titer neutralizing antibodies against HPV18, HPV45 and HPV59 in mice; and their protective effects against HPV18 were comparable to that of HPV18N65 VLP alone and that of the mixed HPV18/HPV45/HPV59 VLP, and were significantly higher than that of HPV45N27 VLP alone and that of HPV59 VLP alone; and their protective effects against HPV45 were comparable to that of HPV45N27 VLP alone and that of the mixed HPV18/HPV45/HPV59 VLP, and were significantly higher than that of HPV18N65 VLP alone and that of HPV59 VLP alone; and their protective effects against HPV59 were comparable to that of HPV59 VLP alone and that of the mixed HPV18/HPV45/HPV59 VLP, and were significantly higher than that of HPV45N27 VLP alone and that of HPV18N65 VLP alone. This showed that H18N65-45T3-59S1 VLP and H18N65-45T4-59S1 VLP had good cross-immunogenicity and cross-protection against HPV18, HPV45 and HPV59, and could be used as effective vaccines for preventing HPV18 infection, HPV45 infection and/or HPV59 infection, and could be used in place of a mixed vaccine comprising HPV18 VLP, HPV45 VLP and HPV59 VLP.

(9) FIGS. 8A-8B show the result of neutralizing antibody titer in mouse serum after vaccination of mice with H18N65-45T4 VLP. FIG. 8A: Group of dose of 10 μg (at an immunizing dose of 10 μg, using aluminum adjuvant); FIG. 8B: Group of dose of 1 μg (at an immunizing dose of 1 μg, using aluminum adjuvant). The result showed that H18N65-45T4 VLP could induce the generation of high-titer neutralizing antibodies against HPV18 in mice, and its protective effect was comparable to that of HPV18N65 VLP alone and that of the mixed HPV18/HPV45 VLP at the same dose, and was significantly superior to that of HPV45N27 VLP alone at the same dose; and it could induce the generation of high-titer neutralizing antibodies against HPV45 in mice, and its protective effect was comparable to that of HPV45N27 VLP alone and that of the mixed HPV18/HPV45 VLP at the same dose, and was significantly superior to that of HPV18N65 VLP alone at the same dose. This showed that H18N65-45T4 VLP had good cross-immunogenicity and cross-protection against HPV18 and HPV45.

(10) FIG. 8C-8D show the result of neutralizing antibody titer in mouse serum after vaccination of mice with H18N65-45T3-59S1 VLP, H18N65-45T4-59S1 VLP, H18N65-45T1-59S5 VLP, H18N65-45T2-59S5 VLP OR H18N65-45T1T3-59S5 VLP. FIG. 8C: Group of dose of 10 μg (at an immunizing dose of 10 μg, using aluminum adjuvant); FIG. 8D: Group of dose of 1 μg (at an immunizing dose of 1 μg, using aluminum adjuvant). The result showed that H18N65-45T3-59S1 VLP, H18N65-45T4-59S1 VLP, H18N65-45T1-59S5 VLP, H18N65-45T2-59S5 VLP and H18N65-45T1T3-59S5 VLP could induce the generation of high-titer neutralizing antibodies against HPV18 in mice, and their protective effects were comparable to that of HPV18N65 VLP alone and that of the mixed HPV18/HPV45/HPV59 VLP at the same dose, and were significantly superior to that of HPV45N27 VLP alone or that of HPV59 VLP alone at the same dose; and H18N65-45T3-59S1 VLP, H18N65-45T4-59S1 VLP, H18N65-45T1-59S5 VLP, H18N65-45T2-59S5 VLP and H18N65-45T1T3-59S5 VLP could induce the generation of high-titer neutralizing antibodies against HPV45 in mice, and their protective effects were comparable to that of HPV45N27 VLP alone and that of the mixed HPV18/HPV45/HPV59 VLP at the same dose, and were significantly superior to that of HPV18N65 VLP alone or that of HPV59 VLP alone at the same dose; and H18N65-45T3-59S1 VLP, H18N65-45T4-59S1 VLP, H18N65-45T1-59S5 VLP, H18N65-45T2-59S5 VLP and H18N65-45T1T3-59S5 VLP could induce the generation of high-titer neutralizing antibodies against HPV59 in mice, and their protective effects were comparable to that of HPV59 VLP alone or that of the mixed HPV18/HPV45/HPV59 VLP at the same dose, and were significantly superior to that of HPV18N65 VLP alone or that of HPV45N27 VLP alone at the same dose. This showed that H18N65-45T3-59S1 VLP, H18N65-45T4-59S1 VLP, H18N65-45T1-59S5 VLP, H18N65-45T2-59S5 VLP and H18N65-45T1T3-59S5 VLP had good cross-immunogenicity and cross-protection against HPV18, HPV45 and HPV59.

SEQUENCE INFORMATION

(11) Some of the sequences involved in the invention are provided in the following Table 1.

(12) TABLE-US-00001 TABLE 1 Description of sequences SEQ ID NO: Description 1 wild type HPV18 L1 protein 2 wild type HPV45 Ll protein 3 wild type HPV59 Ll protein 4 the mutated HPV18 L1 protein comprising Segment 1 of HPV45 L1 protein, H18N65-45T1 5 the mutated HPV18 L1 protein comprising Segment 2 of HPV45 L1 protein, H18N65-45T2 6 the mutated HPV18 L1 protein comprising Segment 3 of HPV45 L1 protein, H18N65-45T3 7 the mutated HPV18 L1 protein comprising Segment 4 of HPV45 L1 protein, H18N65-45T4 8 the mutated HPV18 L1 protein comprising Segment 5 of HPV45 L1 protein, H18N65-45T5 9 the mutated HPV18 L1 protein comprising Segment 3 of HPV45 L1 protein and Segment 1 of HPV59 L1 protein, H18N65-45T3-59S1 10 the mutated HPV18 L1 protein comprising Segment 3 of HPV45 L1 protein and Segment 2 of HPV59 L1 protein, H18N65-45T3-59S2 11 the mutated HPV18 L1 protein comprising Segment 3 of HPV45 L1 protein and Segment 4 of HPV59 L1 protein, H18N65-45T3-59S4 12 the mutated HPV18 L1 protein comprising Segment 3 of HPV45 L1 protein and Segment 5 of HPV59 L1 protein, H18N65-45T3-59S5 13 the mutated HPV18 L1 protein comprising Segment 4 of HPV45 L1 protein and Segment 1 of HPV59 L1 protein, H18N65-45T4-59S1 14 the mutated HPV18 L1 protein comprising Segment 4 of HPV45 L1 protein and Segment 2 of HPV59 L1 protein, H18N65-45T4-59S2 15 the mutated HPV18 L1 protein comprising Segment 4 of HPV45 L1 protein and Segment 3 of HPV59 L1 protein, H18N65-45T4-59S3 16 the mutated HPV18 L1 protein comprising Segment 4 of HPV45 L1 protein and Segment 5 of HPV59 L1 protein, H18N65-45T4-59S5 17 the mutated HPV18 L1 protein comprising Segment 1 of HPV45 L1 protein and Segment 5 of HPV59 L1 protein, H18N65-45T1-59S5 18 the mutated HPV18 L1 protein comprising Segment 2 of HPV45 L1 protein and Segment 5 of HPV59 L1 protein, H18N65-45T2-59S5 19 the mutated HPV18 L1 protein comprising Segment 1 and Segment 3 of HPV45 L1 protein and Segment 5 of HPV59 L1 protein, H18N65-45T1T3-59S5 20 the DNA sequence encoding SEQ ID NO: 1 21 the DNA sequence encoding SEQ ID NO: 2 22 the DNA sequence encoding SEQ ID NO: 3 23 the DNA sequence encoding SEQ ID NO: 4 24 the DNA sequence encoding SEQ ID NO: 5 25 the DNA sequence encoding SEQ ID NO: 6 26 the DNA sequence encoding SEQ ID NO: 7 27 the DNA sequence encoding SEQ ID NO: 8 28 the DNA sequence encoding SEQ ID NO: 9 29 the DNA sequence encoding SEQ ID NO: 10 30 the DNA sequence encoding SEQ ID NO: 11 31 the DNA sequence encoding SEQ ID NO: 12 32 the DNA sequence encoding SEQ ID NO: 13 33 the DNA sequence encoding SEQ ID NO: 14 34 the DNA sequence encoding SEQ ID NO: 15 35 the DNA sequence encoding SEQ ID NO: 16 36 the DNA sequence encoding SEQ ID NO: 17 37 the DNA sequence encoding SEQ ID NO: 18 38 the DNA sequence encoding SEQ ID NO: 19 39 the sequence of the amino acid residues at positions 79-89 of wild type HPV45 L1 protein, Segment 1 of HPV45 L1 protein 40 the sequence of the amino acid residues at positions 142-168 of wild type HPV45 L1 protein, Segment 2 of HPV45 L1 protein 41 the sequence of the amino acid residues at positions 201-209 of wild type HPV45 L1 protein, Segment 3 of HPV45 L1 protein 42 the sequence of the amino acid residues at positions 293-314 of wild type HPV45 L1 protein, Segment 4 of HPV45 L1 protein 43 the sequence of the amino acid residues at positions 51-62 of wild type HPV59 L1 protein, Segment 1 of HPV59 L1 protein 44 the sequence of the amino acid residues at positions 349-360 of wild type HPV59 L1 protein, Segment 5 of HPV59 L1 protein 109 the sequence of the amino acid residues at positions 379-387 of wild type HPV45 L1 protein, Segment 5 of HPV45 L1 protein 110 the sequence of the amino acid residues at positions 122-143 of wild type HPV59 L1 protein, Segment 2 of HPV59 L1 protein 111 the sequence of the amino acid residues at positions 264-290 of wild type HPV59 L1 protein, Segment 4 of HPV59 L1 protein 112 the sequence of the amino acid residues at positions 170-181 of wild type HPV59 L1 protein, Segment 3 of HPV59 L1 protein 113 the HPV18 L1 protein having 65 amino acids truncated at N-terminal, HPV18N65 114 the DNA sequence encoding SEQ ID NO: 113 115 the HPV45 L1 protein having 27 amino acids truncated at N-terminal, HPV45N27 116 the DNA sequence encoding SEQ ID NO: 115

(13) TABLE-US-00002 Sequence 1 (SEQ ID NO: 1): MCLYTRVLILHYHLLPLYGPLYHPQPLPLHSILVYMVHIIICGHYIILF LRNVNVFPIFLQMALWRPSDNTVYLPPPSVARVVNTDDYVTRTSIFYHA GSSRLLTVGNPYFRVPAGGGNKQDIPKVSAYQYRVFRVQLPDPNKFGLP DTSIYNPETQRLVWACAGVEIGRGQPLGVGLSGHPFYNKLDDTESSHAA TSNVSEDVRDNVSVDYKQTQLCILGCAPAIGEHWAKGTACKSRPLSQGD CPPLELKNTVLEDGDMVDTGYGAMDFSTLQDTKCEVPLDICQSICKYPD YLQMSADPYGDSMFFCLRREQLFARHFWNRAGTMGDTVPQSLYIKGTGM RASPGSCVYSPSPSGSIVTSDSQLFNKPYWLHKAQGHNNGVCWHNQLFV TVVDTTRSTNLTICASTQSPVPGQYDATKFKQYSRHVEEYDLQFIFQLC TITLTADVMSYIHSMNSSILEDWNFGVPPPPTTSLVDTYRFVQSVAIAC QKDAAPAENKDPYDKLKFWNVDLKEKFSLDLDQYPLGRKELVQAGLRRK PTIGPRKRSAPSATTASKPAKRVRVRARK

(14) TABLE-US-00003 Sequence 2 (SEQ ID NO: 2): MAHNIIYGHGIIIFLKNVNVFPIFLQMALWRPSDSTVYLPPPSVARVVS TDDYVSRTSIFYHAGSSRLLTVGNPYFRVVPNGAGNKQAVPKVSAYQYR VFRVALPDPNKFGLPDSTIYNPETQRLVWACVGMEIGRGQPLGIGLSGH PFYNKLDDTESAHAATAVITQDVRDNVSVDYKQTQLCILGCVPAIGEHW AKGTLCKPAQLQPGDCPPLELKNTIIEDGDMVDTGYGAMDFSTLQDTKC EVPLDICQSICKYPDYLQMSADPYGDSMFFCLRREQLFARHFWNRAGVM GDTVPTDLYIKGTSANMRETPGSCVYSPSPSGSIITSDSQLFNKPYWLH KAQGHNNGICWHNQLFVTVVDTTRSTNLTLCASTQNPVPSTYDPTKFKQ YSRHVEEYDLQFIFQLCTITLTAEVMSYIHSMNSSILENWNFGVPPPPT TSLVDTYRFVQSVAVTCQKDTTPPEKQDPYDKLKFWTVDLKEKFSSDLD QYPLGRKFLVQAGLRRRPTIGPRKRPAASTSTASTASRPAKRVRIRSKK

(15) TABLE-US-00004 Sequence 3 (SEQ ID NO: 3): MALWRSSDNKVYLPPPSVAKVVSTDEYVTRTSIFYHAGSSRLLTVGHPY FKVPKGGNGRQDVPKVSAYQYRVFRVKLPDPNKFGLPDNTVYDPNSQRL VWACVGVEIGRGQPLGVGLSGHPLYNKLDDTENSHVASAVDTKDTRDNV SVDYKQTQLCIIGCVPAIGEHWTKGTACKPTTVVQGDCPPLELINTPIE DGDMVDTGYGAMDFKLLQDNKSEVPLDICQSICKYPDYLQMSADAYGDS MFFCLRREQVFARHFWNRSGTMGDQLPESLYIKGTDIRANPGSYLYSPS PSGSVVTSDSQLFNKPYWLHKAQGLNNGICWHNQLFLTVVDTTRSTNLS VCASTTSSIPNVYTPTSFKEYARHVEEFDLQFIFQLCKITLTTEVMSYI HNMNTTILEDWNFGVTPPPTASLVDTYRFVQSAAVTCQKDTAPPVKQDP YDKLKFWPVDLKERFSADLDQFPLGRKFLLQLGARPKPTIGPRKRAAPA PTSTPSPKRVKRRKSSRK

(16) TABLE-US-00005 Sequence 4 (SEQ ID NO: 4): MRPSDNTVYLPPPSVARVVNTDDYVTRTSIFYHAGSSRLLTVGNPYFRV VPNGAGNKQAVPKVSAYQYRVFRVQLPDPNKFGLPDTSIYNPETQRLVW ACAGVEIGRGQPLGVGLSGHPFYNKLDDTESSHAATSNVSEDVRDNVSV DYKQTQLCILGCAPAIGEHWAKGTACKSRPLSQGDCPPLELKNTVLEDG DMVDTGYGAMDFSTLQDTKCEVPLDICQSICKYPDYLQMSADPYGDSMF FCLRREQLFARHFWNRAGTMGDTVPQSLYIKGTGMRASPGSCVYSPSPS GSIVTSDSQLFNKPYWLHKAQGHNNGVCWHNQLFVTVVDTTRSTNLTIC ASTQSPVPGQYDATKFKQYSRHVEEYDLQFIFQLCTITLTADVMSYIHS MNSSILEDWNFGVPPPPTTSLVDTYRFVQSVAIACQKDAAPAENKDPYD KLKFWNVDLKEKFSLDLDQYPLGRKFLVQAGLRRKPTIGPRKRSAPSAT TASKPAKRVRVRARK

(17) TABLE-US-00006 Sequence 5 (SEQ ID NO: 5): MRPSDNTVYLPPPSVARVVNTDDYVTRTSIFYHAGSSRLLTVGNPYFRV PAGGGNKQDIPKVSAYQYRVFRVQLPDPNKFGLPDTSIYNPETQRLVWA CAGVEIGRGQPLGIGLSGHPFYNKLDDTESAHAATAVITQDVRDNVSVD YKQTQLCILGCAPAIGEHWAKGTACKSRPLSQGDCPPLELKNTVLEDGD MVDTGYGAMDFSTLQDTKCEVPLDICQSICKYPDYLQMSADPYGDSMFF CLRREQLFARHFWNRAGTMGDTVPQSLYIKGTGMRASPGSCVYSPSPSG SIVTSDSQLFNKPYWLHKAQGHNNGVCWHNQLFVTVVDTTRSTNLTICA STQSPVPGQYDATKFKQYSRHVEEYDLQFIFQLCTITLTADVMSYIHSM NSSILEDWNFGVPPPPTTSLVDTYRFVQSVAIACQKDAAPAENKDPYDK LKFWNVDLKEKFSLDLDQYPLGRKFLVQAGLRRKPTIGPRKRSAPSATT ASKPAKRVRVRARK

(18) TABLE-US-00007 Sequence 6 (SEQ ID NO: 6): MRPSDNTVYLPPPSVARVVNTDDYVTRTSIFYHAGSSRLLTVGNPYFRV PAGGGNKQDIPKVSAYQYRVFRVQLPDPNKFGLPDTSIYNPETQRLVWA CAGVEIGRGQPLGVGLSGHPFYNKLDDTESSHAATSNVSEDVRDNVSVD YKQTQLCILGCAPAIGEHWAKGTLCKPAQLQPGDCPPLELKNTVLEDGD MVDTGYGAMDFSTLQDTKCEVPLDICQSICKYPDYLQMSADPYGDSMFF CLRREQLFARHFWNRAGTMGDTVPQSLYIKGTGMRASPGSCVYSPSPSG SIVTSDSQLFNKPYWLHKAQGHNNGVCWHNQLFVTVVDTTRSTNLTICA STQSPVPGQYDATKFKQYSRHVEEYDLQFIFQLCTITLTADVMSYIHSM NSSILEDWNFGVPPPPTTSLVDTYRFVQSVAIACQKDAAPAENKDPYDK LKFWNVDLKEKFSLDLDQYPLGRKFLVQAGLRRKPTIGPRKRSAPSATT ASKPAKRVRVRARK

(19) TABLE-US-00008 Sequence 7 (SEQ ID NO: 7): MRPSDNTVYLPPPSVARVVNTDDYVTRTSIFYHAGSSRLLTVGNPYFRV PAGGGNKQDIPKVSAYQYRVFRVQLPDPNKFGLPDTSIYNPETQRLVWA CAGVEIGRGQPLGVGLSGHPFYNKLDDTESSHAATSNVSEDVRDNVSVD YKQTQLCILGCAPAIGEHWAKGTACKSRPLSQGDCPPLELKNTVLEDGD MVDTGYGAMDFSTLQDTKCEVPLDICQSICKYPDYLQMSADPYGDSMFF CLRREQLFARHFWNRAGVMGDTVPTDLYIKGTSANMRETPGSCVYSPSP SGSIVTSDSQLFNKPYWLHKAQGHNNGVCWHNQLFVTVVDTTRSTNLTI CASTQSPVPGQYDATKFKQYSRHVEEYDLQFIFQLCTITLTADVMSYIH SMNSSILEDWNFGVPPPPTTSLVDTYRFVQSVAIACQKDAAPAENKDPY DKLKFWNVDLKEKFSLDLDQYPLGRKFLVQAGLRRKPTIGPRKRSAPSA TTASKPAKRVRVRARK

(20) TABLE-US-00009 Sequence 8(SEQ ID NO: 8):  MRPSDNTVYLPPPSVARVVNTDDYVTRTSIFYHAGSSRLLTVGNPYFRV  PAGGGNKQDIPKVSAYQYRVFRVQLPDPNKFGLPDTSIYNPETQRLVWA CAGVEIGRGQPLGVGLSGHPFYNKLDDTESSHAATSNVSEDVRDNVSVD YKQTQLCILGCAPAIGEHWAKGTACKSRPLSQGDCPPLELKNTVLEDGD MVDTGYGAMDFSTLQDTKCEVPLDICQSICKYPDYLQMSADPYGDSMFF CLRREQLFARHFWNRAGTMGDTVPQSLYIKGTGMRASPGSCVYSPSPSG SIVTSDSQLFNKPYWLHKAQGHNNGVCWHNQLFVTVVDTTRSTNLTICA STQNPVPSTYDPTKFKQYSRHVEEYDLQFIFQLCTITLTADVMSYIHSM  NSSILEDWNFGVPPPPTTSLVDTYRFVQSVAIACQKDAAPAENKDPYDK LKFWNVDLKEKFSLDLDQYPLGRKFLVQAGLRRKPTIGPRKRSAPSATT ASKPAKRVRVRARK 

(21) TABLE-US-00010 Sequence 9 (SEQ ID NO: 9):  MRPSDNTVYLPPPSVARVVNTDDYVTRTSIFYHAGSSRLLTVGNPYFKV PKGGNGRQDVPKVSAYQYRVFRVQLPDPNKFGLPDTSIYNPETQRLVWA CAGVEIGRGQPLGVGLSGHPFYNKLDDTESSHAATSNVSEDVRDNVSVD YKQTQLCILGCAPAIGEHWAKGTLCKPAQLQPGDCPPLELKNTVLEDGD MVDTGYGAMDFSTLQDTKCEVPLDICQSICKYPDYLQMSADPYGDSMFF CLRREQLFARHFWNRAGTMGDTVPQSLYIKGTGMRASPGSCVYSPSPSG SIVTSDSQLFNKPYWLHKAQGHNNGVCWHNQLFVTVVDTTRSTNLTICA STQSPVPGQYDATKFKQYSRHVEEYDLQFIFQLCTITLTADVMSYIHSM NSSILEDWNFGVPPPPTTSLVDTYRFVQSVAIACQKDAAPAENKDPYDK LKFWNVDLKEKFSLDLDQYPLGRKFLVQAGLRRKPTIGPRKRSAPSATT ASKPAKRVRVRARK

(22) TABLE-US-00011 Sequence 10 (SEQ ID NO: 10):  MRPSDNTVYLPPPSVARVVNTDDYVTRTSIFYHAGSSRLLTVGNPYFRV PAGGGNKQDIPKVSAYQYRVFRVQLPDPNKFGLPDTSIYNPETQRLVWA CAGVEIGRGQPLGVGLSGHPLYNKLDDTENSHVASAVDTKDTRDNVSVD YKQTQLCILGCAPAIGEHWAKGTLCKPAQLQPGDCPPLELKNTVLEDGD MVDTGYGAMDFSTLQDTKCEVPLDICQSICKYPDYLQMSADPYGDSMFF CLRREQLFARHFWNRAGTMGDTVPQSLYIKGTGMRASPGSCVYSPSPSG SIVTSDSQLFNKPYWLHKAQGHNNGVCWHNQLFVTVVDTTRSTNLTICA STQSPVPGQYDATKFKQYSRHVEEYDLQFIFQLCTITLTADVMSYIHSM NSSILEDWNFGVPPPPTTSLVDTYRFVQSVAIACQKDAAPAENKDPYDK LKFWNVDLKEKFSLDLDQYPLGRKFLVQAGLRRKPTIGPRKRSAPSATT ASKPAKRVRVRARK

(23) TABLE-US-00012 Sequence 11 (SEQ ID NO: 11):  MRPSDNTVYLPPPSVARVVNTDDYVTRTSIFYHAGSSRLLTVGNPYFRV PAGGGNKQDIPKVSAYQYRVFRVQLPDPNKFGLPDTSIYNPETQRLVWA CAGVEIGRGQPLGVGLSGHPFYNKLDDTESSHAATSNVSEDVRDNVSVD YKQTQLCILGCAPAIGEHWAKGTLCKPAQLQPGDCPPLELKNTVLEDGD MVDTGYGAMDFSTLQDTKCEVPLDICQSICKYPDYLQMSADPYGDSMFF CLRREQLFARHFWNRSGTMGDQLPESLYIKGTDIRANPGSYLYSPSPSG SIVTSDSQLFNKPYWLHKAQGHNNGVCWHNQLFVTVVDTTRSTNLTICA STQSPVPGQYDATKFKQYSRHVEEYDLQFIFQLCTITLTADVMSYIHSM NSSILEDWNFGVPPPPTTSLVDTYRFVQSVAIACQKDAAPAENKDPYDK LKFWNVDLKEKFSLDLDQYPLGRKFLVQAGLRRKPTIGPRKRSAPSATT ASKPAKRVRVRARK

(24) TABLE-US-00013 Sequence 12 (SEQ ID NO: 12):  MRPSDNTVYLPPPSVARVVNTDDYVTRTSIFYHAGSSRLLTVGNPYFRV PAGGGNKQDIPKVSAYQYRVFRVQLPDPNKFGLPDTSIYNPETQRLVWA CAGVEIGRGQPLGVGLSGHPFYNKLDDTESSHAATSNVSEDVRDNVSVD YKQTQLCILGCAPAIGEHWAKGTLCKPAQLQPGDCPPLELKNTVLEDGD MVDTGYGAMDFSTLQDTKCEVPLDICQSICKYPDYLQMSADPYGDSMFF CLRREQLFARHFWNRAGTMGDTVPQSLYIKGTGMRASPGSCVYSPSPSG SIVTSDSQLFNKPYWLHKAQGHNNGVCWHNQLFVTVVDTTRSTNLTICA STTSSIPNVYTPTSFKQYSRHVEEYDLQFIFQLCTITLTADVMSYIHSM NSSILEDWNFGVPPPPTTSLVDTYRFVQSVAIACQKDAAPAENKDPYDK LKFWNVDLKEKFSLDLDQYPLGRKFLVQAGLRRKPTIGPRKRSAPSATT ASKPAKRVRVRARK

(25) TABLE-US-00014 Sequence 13 (SEQ ID NO: 13):  MRPSDNTVYLPPPSVARVVNTDDYVTRTSIFYHAGSSRLLTVGNPYFKV PKGGNGRQDVPKVSAYQYRVFRVQLPDPNKFGLPDTSIYNPETQRLVWA CAGVEIGRGQPLGVGLSGHPFYNKLDDTESSHAATSNVSEDVRDNVSVD YKQTQLCILGCAPAIGEHWAKGTACKSRPLSQGDCPPLELKNTVLEDGD MVDTGYGAMDFSTLQDTKCEVPLDICQSICKYPDYLQMSADPYGDSMFF CLRREQLFARHFWNRAGVMGDTVPTDLYIKGTSANMRETPGSCVYSPSP SGSIVTSDSQLFNKPYWLHKAQGHNNGVCWHNQLFVTVVDTTRSTNLTI CASTQSPVPGQYDATKFKQYSRHVEEYDLQFIFQLCTITLTADVMSYIH SMNSSILEDWNFGVPPPPTTSLVDTYRFVQSVAIACQKDAAPAENKDPY DKLKFWNVDLKEKFSLDLDQYPLGRKFLVQAGLRRKPTIGPRKRSAPSA TTASKPAKRVRVRARK

(26) TABLE-US-00015 Sequence 14 (SEQ ID NO: 14):  MRPSDNTVYLPPPSVARVVNTDDYVTRTSIFYHAGSSRLLTVGNPYFRV PAGGGNKQDIPKVSAYQYRVFRVQLPDPNKFGLPDTSIYNPETQRLVWA CAGVEIGRGQPLGVGLSGHPLYNKLDDTENSHVASAVDTKDTRDNVSVD YKQTQLCILGCAPAIGEHWAKGTACKSRPLSQGDCPPLELKNTVLEDGD MVDTGYGAMDFSTLQDTKCEVPLDICQSICKYPDYLQMSADPYGDSMFF CLRREQLFARHFWHNQLFVTVVDTTRSTNLTICASTQSPVPGQYDATKF KQYSRHVEEYDLQFIFQLCTITLTADVMSYIHSMNSSILEDWNFGVPPP PTTSLVDTYRFVQSVAIACQKDAAPAENKDPYDKLKFWNVDLKEKFSLD LDQYPLGRKFLVQAGLRRKPTIGPRKRSAPSATTASKPAKRVRVRARK

(27) TABLE-US-00016 Sequence 15 (SEQ ID NO: 15):  MRPSDNTVYLPPPSVARVVNTDDYVTRTSIFYHAGSSRLLTVGNPYFRV PAGGGNKQDIPKVSAYQYRVFRVQLPDPNKFGLPDTSIYNPETQRLVWA CAGVEIGRGQPLGVGLSGHPFYNKLDDTESSHAATSNVSEDVRDNVSVD YKQTQLCILGCAPAIGEHWTKGTACKPTTVVQGDCPPLELKNTVLEDGD MVDTGYGAMDFSTLQDTKCEVPLDICQSICKYPDYLQMSADPYGDSMFF CLRREQLFARHFWNRAGVMGDTVPTDLYIKGTSANMRETPGSCVYSPSP SGSIVTSDSQLFNKPYWLHKAQGHNNGVCWHNQLFVTVVDTTRSTNLTI CASTQSPVPGQYDATKFKQYSRHVEEYDLQFIFQLCTITLTADVMSYIH SMNSSILEDWNFGVPPPPTTSLVDTYRFVQSVAIACQKDAAPAENKDPY DKLKFWNVDLKEKFSLDLDQYPLGRKFLVQAGLRRKPTIGPRKRSAPSA TTASKPAKRVRVRARK

(28) TABLE-US-00017 Sequence 16 (SEQ ID NO: 16):  MRPSDNTVYLPPPSVARVVNTDDYVTRTSIFYHAGSSRLLTVGNPYFRV PAGGGNKQDIPKVSAYQYRVFRVQLPDPNKFGLPDTSIYNPETQRLVWA CAGVEIGRGQPLGVGLSGHPFYNKLDDTESSHAATSNVSEDVRDNVSVD YKQTQLCILGCAPAIGEHWAKGTACKSRPLSQGDCPPLELKNTVLEDGD MVDTGYGAMDFSTLQDTKCEVPLDICQSICKYPDYLQMSADPYGDSMFF CLRREQLFARHFWNRAGVMGDTVPTDLYIKGTSANMRETPGSCVYSPSP SGSIVTSDSQLFNKPYWLHKAQGHNNGVCWHNQLFVTVVDTTRSTNLTI CASTTSSIPNVYTPTSFKQYSRHVEEYDLQFIFQLCTITLTADVMSYIH SMNSSILEDWNFGVPPPPTTSLVDTYRFVQSVAIACQKDAAPAENKDPY DKLKFWNVDLKEKFSLDLDQYPLGRKFLVQAGLRRKPTIGPRKRSAPSA TTASKPAKRVRVRARK

(29) TABLE-US-00018 Sequence 17 (SEQ ID NO: 17):  MRPSDNTVYLPPPSVARVVNTDDYVTRTSIFYHAGSSRLLTVGNPYFRV VPNGAGNKQAVPKVSAYQYRVFRVQLPDPNKFGLPDTSIYNPETQRLVW ACAGVEIGRGQPLGVGLSGHPFYNKLDDTESSHAATSNVSEDVRDNVSV DYKQTQLCILGCAPAIGEHWAKGTACKSRPLSQGDCPPLELKNTVLEDG DMVDTGYGAMDFSTLQDTKCEVPLDICQSICKYPDYLQMSADPYGDSMF FCLRREQLFARHFWNRAGTMGDTVPQSLYIKGTGMRASPGSCVYSPSPS GSIVTSDSQLFNKPYWLHKAQGHNNGVCWHNQLFVTVVDTTRSTNLTIC ASTTSSIPNVYTPTSFKQYSRHVEEYDLQFIFQLCTITLTADVMSYIHS MNSSILEDWNFGVPPPPTTSLVDTYRFVQSVAIACQKDAAPAENKDPYD KLKFWNVDLKEKFSLDLDQYPLGRKFLVQAGLRRKPTIGPRKRSAPSAT TASKPAKRVRVRARK

(30) TABLE-US-00019 Sequence 18 (SEQ ID NO: 18):  MRPSDNTVYLPPPSVARVVNTDDYVTRTSIFYHAGSSRLLTVGNPYFRV PAGGGNKQDIPKVSAYQYRVFRVQLPDPNKFGLPDTSIYNPETQRLVWA CAGVEIGRGQPLGIGLSGHPFYNKLDDTESAHAATAVITQDVRDNVSVD YKQTQLCILGCAPAIGEHWAKGTACKSRPLSQGDCPPLELKNTVLEDGD MVDTGYGAMDFSTLQDTKCEVPLDICQSICKYPDYLQMSADPYGDSMFF CLRREQLFARHFWNRAGTMGDTVPQSLYIKGTGMRASPGSCVYSPSPSG SIVTSDSQLFNKPYWLHKAQGHNNGVCWHNQLFVTVVDTTRSTNLTICA STTSSIPNVYTPTSFKQYSRHVEEYDLQFIFQLCTITLTADVMSYIHSM NSSILEDWNFGVPPPPTTSLVDTYRFVQSVAIACQKDAAPAENKDPYDK LKFWNVDLKEKFSLDLDQYPLGRKFLVQAGLRRKPTIGPRKRSAPSATT ASKPAKRVRVRARK

(31) TABLE-US-00020 Sequence 19 (SEQ ID NO: 19):  MRPSDNTVYLPPPSVARVVNTDDYVTRTSIFYHAGSSRLLTVGNPYFRV VPNGAGNKQAVPKVSAYQYRVFRVQLPDPNKFGLPDTSIYNPETQRLVW ACAGVEIGRGQPLGVGLSGHPFYNKLDDTESSHAATSNVSEDVRDNVSV DYKQTQLCILGCAPAIGEHWAKGTLCKPAQLQPGDCPPLELKNTVLEDG DMVDTGYGAMDFSTLQDTKCEVPLDICQSICKYPDYLQMSADPYGDSMF FCLRREQLFARHFWNRAGTMGDTVPQSLYIKGTGMRASPGSCVYSPSPS GSIVTSDSQLFNKPYWLHKAQGHNNGVCWHNQLFVTVVDTTRSTNLTIC ASTTSSIPNVYTPTSFKQYSRHVEEYDLQFIFQLCTITLTADVMSYIHS MNSSILEDWNFGVPPPPTTSLVDTYRFVQSVAIACQKDAAPAENKDPYD KLKFWNVDLKEKFSLDLDQYPLGRKFLVQAGLRRKPTIGPRKRSAPSAT TASKPAKRVRVRARK

(32) TABLE-US-00021 Sequence 20 (SEQ ID NO: 20):  ATGTGCCTGTATACACGGGTCCTGATATTACATTACCATCTACTACCTC TGTATGGCCCATTGTATCACCCACAGCCCCTGCCTCTACACAGTATATT GGTATACATGGTACACATTATTATTTGTGGCCATTATATTATTTTATTC CTAAGAAACGTAAACGTGTTCCCTATTTTTTTGCAGATGGCTTTGTGGC GGCCTAGTGACAATACCGTATATCTTCCACCTCCTTCTGTGGCAAGAGT TGTAAATACCGATGATTACGTGACTCGCACAAGCATATTTTATCATGCT GGCAGCTCTAGATTATTAACTGTTGGTAATCCATATTTTAGGGTTCCTG CAGGTGGTGGCAATAAGCAGGATATTCCTAAGGTTTCTGCATACCAATA TAGAGTATTTAGGGTGCAGTTACCTGACCCAAATAAATTTGGTTTACCT GATACTAGTATTTATAATCCTGAAACACAACGTTTAGTGTGGGCCTGTG CTGGAGTGGAAATTGGCCGTGGTCAGCCTTTAGGTGTTGGCCTTAGTGG GCATCCATTTTATAATAAATTAGATGACACTGAAAGTTCCCATGCCGCC ACGTCTAATGTTTCTGAGGACGTTAGGGACAATGTGTCTGTAGATTATA AGCAGACACAGTTATGTATTTTGGGCTGTGCCCCTGCTATTGGGGAACA CTGGGCTAAAGGCACTGCTTGTAAATCGCGTCCTTTATCACAGGGCGAT TGCCCCCCTTTAGAACTTAAAAACACAGTTTTGGAAGATGGTGATATGG TAGATACTGGATATGGTGCCATGGACTTTAGTACATTGCAAGATACTAA ATGTGAGGTACCATTGGATATTTGTCAGTCTATTTGTAAATATCCTGAT TATTTACAAATGTCTGCAGATCCTTATGGGGATTCCATGTTTTTTTGCT TACGGCGTGAGCAGCTTTTTGCTAGGCATTTTTGGAATAGAGCAGGTAC TATGGGTGACACTGTGCCTCAATCCTTATATATTAAAGGCACAGGTATG CGTGCTTCACCTGGCAGCTGTGTGTATTCTCCCTCTCCAAGTGGCTCTA TTGTTACCTCTGACTCCCAGTTGTTTAATAAACCATATTGGTTACATAA GGCACAGGGTCATAACAATGGTGTTTGCTGGCATAATCAATTATTTGTT ACTGTGGTAGATACCACTCGCAGTACCAATTTAACAATATGTGCTTCTA CACAGTCTCCTGTACCTGGGCAATATGATGCTACCAAATTTAAGCAGTA TAGCAGACATGTTGAGGAATATGATTTGCAGTTTATTTTTCAGTTGTGT ACTATTACTTTAACTGCAGATGTTATGTCCTATATTCATAGTATGAATA GCAGTATTTTAGAGGATTGGAACTTTGGTGTTCCCCCCCCGCCAACTAC TAGTTTGGTGGATACATATCGTTTTGTACAATCTGTTGCTATTGCCTGT CAAAAGGATGCTGCACCGGCTGAAAATAAGGATCCCTATGATAAGTTAA AGTTTTGGAATGTGGATTTAAAGGAAAAGTTTTCTTTAGACTTAGATCA ATATCCCCTTGGACGTAAATTTTTGGTTCAGGCTGGATTGCGTCGCAAG CCCACCATAGGCCCTCGCAAACGTTCTGCTCCATCTGCCACTACGGCTT CTAAACCTGCCAAGCGTGTGCGTGTACGTGCCAGGAAGTAA

(33) TABLE-US-00022 Sequence 21 (SEQ ID NO: 21):  ATGGCACACAATATTATTTATGGCCATGGTACTATTATTTTCCTAAAAA ACGTAAACGTATTCCCTATTTTTTTGCAGATGGCCCTGTGGAGGCCCAG CGACAGCACCGTGTACCTGCCCCCCCCCAGCGTGGCCAGGGTGGTGAGC ACCGACGACTACGTGAGCAGGACCAGCATCTTCTACCACGCCGGCAGCA GCAGGCTGCTGACCGTGGGCAACCCCTACTTCAGGGTGGTGCCCAACGG CGCCGGCAACAAGCAGGCCGTGCCCAAGGTGAGCGCCTACCAGTACAGG GTGTTCAGGGTGGCCCTGCCCGACCCCAACAAGTTCGGCCTGCCCGACA GCACCATCTACAACCCCGAGACCCAGAGGCTGGTGTGGGCCTGCGTGGG CATGGAGATCGGCAGGGGCCAGCCCCTGGGCATCGGCCTGAGCGGCCAC CCCTTCTACAACAAGCTGGACGACACCGAGAGCGCCCACGCCGCCACCG CCGTGATCACCCAGGACGTGAGGGACAACGTGAGCGTGGACTACAAGCA GACCCAGCTGTGCATCCTGGGCTGCGTGCCCGCCATCGGCGAGCACTGG GCCAAGGGCACCCTGTGCAAGCCCGCCCAGCTGCAGCCCGGCGACTGCC CCCCCCTGGAGCTGAAGAACACCATCATCGAGGACGGCGACATGGTGGA CACCGGCTACGGCGCCATGGACTTCAGCACCCTGCAGGACACCAAGTGC GAGGTGCCCCTGGACATCTGCCAGAGCATCTGCAAGTACCCCGACTACC TGCAGATGAGCGCCGACCCCTACGGCGACAGCATGTTCTTCTGCCTGAG GAGGGAGCAGCTGTTCGCCAGGCACTTCTGGAACAGGGCCGGCGTGATG GGCGACACCGTGCCCACCGACCTGTACATCAAGGGCACCAGCGCCAACA TGAGGGAGACCCCCGGCAGCTGCGTGTACAGCCCCAGCCCCAGCGGCAG CATCATCACCAGCGACAGCCAGCTGTTCAACAAGCCCTACTGGCTGCAC AAGGCCCAGGGCCACAACAACGGCATCTGCTGGCACAACCAGCTGTTCG TGACCGTGGTGGACACCACCAGGAGCACCAACCTGACCCTGTGCGCCAG CACCCAGAACCCCGTGCCCAGCACCTACGACCCCACCAAGTTCAAGCAG TACAGCAGGCACGTGGAGGAGTACGACCTGCAGTTCATCTTCCAGCTGT GCACCATCACCCTGACCGCCGAGGTGATGAGCTACATCCACAGCATGAA CAGCAGCATCCTGGAGAACTGGAACTTCGGCGTGCCCCCCCCCCCCACC ACCAGCCTGGTGGACACCTACAGGTTCGTGCAGAGCGTGGCCGTGACCT GCCAGAAGGACACCACCCCCCCCGAGAAGCAGGACCCCTACGACAAGCT GAAGTTCTGGACCGTGGACCTGAAGGAGAAGTTCAGCAGCGACCTGGAC CAGTACCCCCTGGGCAGGAAGTTCCTGGTGCAGGCCGGCCTGAGGAGGA GGCCCACCATCGGCCCCAGGAAGAGGCCCGCCGCCAGCACCAGCACCGC CAGCACCGCCAGCAGGCCCGCCAAGAGGGTGAGGATCAGGAGCAAGAAG TGA

(34) TABLE-US-00023 Sequence 22 (SEQ ID NO: 22):  ATGGCCCTGTGGAGGAGCAGCGACAACAAGGTGTACCTGCCCCCCCCCA GCGTGGCCAAGGTGGTGAGCACCGACGAGTACGTGACCAGGACCAGCAT CTTCTACCACGCCGGCAGCAGCAGGCTGCTGACCGTGGGCCACCCCTAC TTCAAGGTGCCCAAGGGCGGCAACGGCAGGCAGGACGTGCCCAAGGTGA GCGCCTACCAGTACAGGGTGTTCAGGGTGAAGCTGCCCGACCCCAACAA GTTCGGCCTGCCCGACAACACCGTGTACGACCCCAACAGCCAGAGGCTG GTGTGGGCCTGCGTGGGCGTGGAGATCGGCAGGGGCCAGCCCCTGGGCG TGGGCCTGAGCGGCCACCCCCTGTACAACAAGCTGGACGACACCGAGAA CAGCCACGTGGCCAGCGCCGTGGACACCAAGGACACCAGGGACAACGTG AGCGTGGACTACAAGCAGACCCAGCTGTGCATCATCGGCTGCGTGCCCG CCATCGGCGAGCACTGGACCAAGGGCACCGCCTGCAAGCCCACCACCGT GGTGCAGGGCGACTGCCCCCCCCTGGAGCTGATCAACACCCCCATCGAG GACGGCGACATGGTGGACACCGGCTACGGCGCCATGGACTTCAAGCTGC TGCAGGACAACAAGAGCGAGGTGCCCCTGGACATCTGCCAGAGCATCTG CAAGTACCCCGACTACCTGCAGATGAGCGCCGACGCCTACGGCGACAGC ATGTTCTTCTGCCTGAGGAGGGAGCAGGTGTTCGCCAGGCACTTCTGGA ACAGGAGCGGCACCATGGGCGACCAGCTGCCCGAGAGCCTGTACATCAA GGGCACCGACATCAGGGCCAACCCCGGCAGCTACCTGTACAGCCCCAGC CCCAGCGGCAGCGTGGTGACCAGCGACAGCCAGCTGTTCAACAAGCCCT ACTGGCTGCACAAGGCCCAGGGCCTGAACAACGGCATCTGCTGGCACAA CCAGCTGTTCCTGACCGTGGTGGACACCACCAGGAGCACCAACCTGAGC GTGTGCGCCAGCACCACCAGCAGCATCCCCAACGTGTACACCCCCACCA GCTTCAAGGAGTACGCCAGGCACGTGGAGGAGTTCGACCTGCAGTTCAT CTTCCAGCTGTGCAAGATCACCCTGACCACCGAGGTGATGAGCTACATC CACAACATGAACACCACCATCCTGGAGGACTGGAACTTCGGCGTGACCC CCCCCCCCACCGCCAGCCTGGTGGACACCTACAGGTTCGTGCAGAGCGC CGCCGTGACCTGCCAGAAGGACACCGCCCCCCCCGTGAAGCAGGACCCC TACGACAAGCTGAAGTTCTGGCCCGTGGACCTGAAGGAGAGGTTCAGCG CCGACCTGGACCAGTTCCCCCTGGGCAGGAAGTTCCTGCTGCAGCTGGG CGCCAGGCCCAAGCCCACCATCGGCCCCAGGAAGAGGGCCGCCCCCGCC CCCACCAGCACCCCCAGCCCCAAGAGGGTGAAGAGGAGGAAGAGCAGCA GGAAGTGA

(35) TABLE-US-00024 Sequence 23 (SEQ ID NO: 23): ATGCGGCCTAGTGACAATACCGTATATCTTCCACCTCCTTCTGTGGCAA GAGTTGTAAATACCGATGATTACGTGACTAGGACCAGCATCTTCTACCA CGCCGGCAGCAGCAGGCTGCTGACCGTGGGCAACCCCTACTTCAGGGTG GTGCCCAACGGCGCCGGCAACAAGCAGGCCGTGCCCAAGGTGAGCGCCT ACCAGTACAGGGTGTTCAGGGTGCAGTTACCTGACCCAAATAAATTTGG TTTACCTGATACTAGTATTTATAATCCTGAAACACAACGTTTAGTGTGG GCCTGTGCTGGAGTGGAAATTGGCCGTGGTCAGCCTTTAGGTGTTGGCC TTAGTGGGCATCCATTTTATAATAAATTAGATGACACTGAAAGTTCCCA TGCCGCCACGTCTAATGTTTCTGAGGACGTTAGGGACAATGTGTCTGTA GATTATAAGCAGACACAGTTATGTATTTTGGGCTGTGCCCCTGCTATTG GGGAACACTGGGCTAAAGGCACTGCTTGTAAATCGCGTCCTTTATCACA GGGCGATTGCCCCCCTTTAGAACTTAAAAACACAGTTTTGGAAGATGGT GATATGGTAGATACTGGATATGGTGCCATGGACTTTAGTACATTGCAAG ATACTAAATGTGAGGTACCATTGGATATTTGTCAGTCTATTTGTAAATA TCCTGATTATTTACAAATGTCTGCAGATCCTTATGGGGATTCCATGTTT TTTTGCTTACGGCGTGAGCAGCTTTTTGCTAGGCATTTTTGGAATAGAG CAGGTACTATGGGTGACACTGTGCCTCAATCCTTATATATTAAAGGCAC AGGTATGCGTGCTTCACCTGGCAGCTGTGTGTATTCTCCCTCTCCAAGT GGCTCTATTGTTACCTCTGACTCCCAGTTGTTTAATAAACCATATTGGT TACATAAGGCACAGGGTCATAACAATGGTGTTTGCTGGCATAATCAATT ATTTGTTACTGTGGTAGATACCACTCGCAGTACCAATTTAACAATATGT GCTTCTACACAGTCTCCTGTACCTGGGCAATATGATGCTACCAAATTTA AGCAGTATAGCAGACATGTTGAGGAATATGATTTGCAGTTTATTTTTCA GTTGTGTACTATTACTTTAACTGCAGATGTTATGTCCTATATTCATAGT ATGAATAGCAGTATTTTAGAGGATTGGAACTTTGGTGTTCCCCCCCCGC CAACTACTAGTTTGGTGGATACATATCGTTTTGTACAATCTGTTGCTAT TGCCTGTCAAAAGGATGCTGCACCGGCTGAAAATAAGGATCCCTATGAT AAGTTAAAGTTTTGGAATGTGGATTTAAAGGAAAAGTTTTCTTTAGACT TAGATCAATATCCCCTTGGACGTAAATTTTTGGTTCAGGCTGGATTGCG TCGCAAGCCCACCATAGGCCCTCGCAAACGTTCTGCTCCATCTGCCACT ACGGCTTCTAAACCTGCCAAGCGTGTGCGTGTACGTGCCAGGAAGTAA

(36) TABLE-US-00025 Sequence 24 (SEQ ID NO: 24): ATGCGGCCTAGTGACAATACCGTATATCTTCCACCTCCTTCTGTGGCAA GAGTTGTAAATACCGATGATTACGTGACTCGCACAAGCATATTTTATCA TGCTGGCAGCTCTAGATTATTAACTGTTGGTAATCCATATTTTAGGGTT CCTGCAGGTGGTGGCAATAAGCAGGATATTCCTAAGGTTTCTGCATACC AATATAGAGTATTTAGGGTGCAGTTACCTGACCCAAATAAATTTGGTTT ACCTGATACTAGTATTTATAATCCTGAAACACAACGTTTAGTGTGGGCC TGTGCTGGAGTGGAGATCGGCAGGGGCCAGCCCCTGGGCATCGGCCTGA GCGGCCACCCCTTCTACAACAAGCTGGACGACACCGAGAGCGCCCACGC CGCCACCGCCGTGATCACCCAGGACGTGAGGGACAACGTGAGCGTGGAC TACAAGCAGACCCAGCTGTGCATCCTGGGCTGCGCCCCTGCTATTGGGG AACACTGGGCTAAAGGCACTGCTTGTAAATCGCGTCCTTTATCACAGGG CGATTGCCCCCCTTTAGAACTTAAAAACACAGTTTTGGAAGATGGTGAT ATGGTAGATACTGGATATGGTGCCATGGACTTTAGTACATTGCAAGATA CTAAATGTGAGGTACCATTGGATATTTGTCAGTCTATTTGTAAATATCC TGATTATTTACAAATGTCTGCAGATCCTTATGGGGATTCCATGTTTTTT TGCTTACGGCGTGAGCAGCTTTTTGCTAGGCATTTTTGGAATAGAGCAG GTACTATGGGTGACACTGTGCCTCAATCCTTATATATTAAAGGCACAGG TATGCGTGCTTCACCTGGCAGCTGTGTGTATTCTCCCTCTCCAAGTGGC TCTATTGTTACCTCTGACTCCCAGTTGTTTAATAAACCATATTGGTTAC ATAAGGCACAGGGTCATAACAATGGTGTTTGCTGGCATAATCAATTATT TGTTACTGTGGTAGATACCACTCGCAGTACCAATTTAACAATATGTGCT TCTACACAGTCTCCTGTACCTGGGCAATATGATGCTACCAAATTTAAGC AGTATAGCAGACATGTTGAGGAATATGATTTGCAGTTTATTTTTCAGTT GTGTACTATTACTTTAACTGCAGATGTTATGTCCTATATTCATAGTATG AATAGCAGTATTTTAGAGGATTGGAACTTTGGTGTTCCCCCCCCGCCAA CTACTAGTTTGGTGGATACATATCGTTTTGTACAATCTGTTGCTATTGC CTGTCAAAAGGATGCTGCACCGGCTGAAAATAAGGATCCCTATGATAAG TTAAAGTTTTGGAATGTGGATTTAAAGGAAAAGTTTTCTTTAGACTTAG ATCAATATCCCCTTGGACGTAAATTTTTGGTTCAGGCTGGATTGCGTCG CAAGCCCACCATAGGCCCTCGCAAACGTTCTGCTCCATCTGCCACTACG GCTTCTAAACCTGCCAAGCGTGTGCGTGTACGTGCCAGGAAGTAA

(37) TABLE-US-00026 Sequence 25 (SEQ ID NO: 25): ATGCGGCCTAGTGACAATACCGTATATCTTCCACCTCCTTCTGTGGCAA GAGTTGTAAATACCGATGATTACGTGACTCGCACAAGCATATTTTATCA TGCTGGCAGCTCTAGATTATTAACTGTTGGTAATCCATATTTTAGGGTT CCTGCAGGTGGTGGCAATAAGCAGGATATTCCTAAGGTTTCTGCATACC AATATAGAGTATTTAGGGTGCAGTTACCTGACCCAAATAAATTTGGTTT ACCTGATACTAGTATTTATAATCCTGAAACACAACGTTTAGTGTGGGCC TGTGCTGGAGTGGAAATTGGCCGTGGTCAGCCTTTAGGTGTTGGCCTTA GTGGGCATCCATTTTATAATAAATTAGATGACACTGAAAGTTCCCATGC CGCCACGTCTAATGTTTCTGAGGACGTTAGGGACAATGTGTCTGTAGAT TATAAGCAGACACAGTTATGTATTTTGGGCTGTGCCCCCGCCATCGGCG AGCACTGGGCCAAGGGCACCCTGTGCAAGCCCGCCCAGCTGCAGCCCGG CGACTGCCCCCCCCTGGAGCTGAAGAACACCGTTTTGGAAGATGGTGAT ATGGTAGATACTGGATATGGTGCCATGGACTTTAGTACATTGCAAGATA CTAAATGTGAGGTACCATTGGATATTTGTCAGTCTATTTGTAAATATCC TGATTATTTACAAATGTCTGCAGATCCTTATGGGGATTCCATGTTTTTT TGCTTACGGCGTGAGCAGCTTTTTGCTAGGCATTTTTGGAATAGAGCAG GTACTATGGGTGACACTGTGCCTCAATCCTTATATATTAAAGGCACAGG TATGCGTGCTTCACCTGGCAGCTGTGTGTATTCTCCCTCTCCAAGTGGC TCTATTGTTACCTCTGACTCCCAGTTGTTTAATAAACCATATTGGTTAC ATAAGGCACAGGGTCATAACAATGGTGTTTGCTGGCATAATCAATTATT TGTTACTGTGGTAGATACCACTCGCAGTACCAATTTAACAATATGTGCT TCTACACAGTCTCCTGTACCTGGGCAATATGATGCTACCAAATTTAAGC AGTATAGCAGACATGTTGAGGAATATGATTTGCAGTTTATTTTTCAGTT GTGTACTATTACTTTAACTGCAGATGTTATGTCCTATATTCATAGTATG AATAGCAGTATTTTAGAGGATTGGAACTTTGGTGTTCCCCCCCCGCCAA CTACTAGTTTGGTGGATACATATCGTTTTGTACAATCTGTTGCTATTGC CTGTCAAAAGGATGCTGCACCGGCTGAAAATAAGGATCCCTATGATAAG TTAAAGTTTTGGAATGTGGATTTAAAGGAAAAGTTTTCTTTAGACTTAG ATCAATATCCCCTTGGACGTAAATTTTTGGTTCAGGCTGGATTGCGTCG CAAGCCCACCATAGGCCCTCGCAAACGTTCTGCTCCATCTGCCACTACG GCTTCTAAACCTGCCAAGCGTGTGCGTGTACGTGCCAGGAAGTAA

(38) TABLE-US-00027 Sequence 26 (SEQ ID NO: 26): ATGCGGCCTAGTGACAATACCGTATATCTTCCACCTCCTTCTGTGGCAA GAGTTGTAAATACCGATGATTACGTGACTCGCACAAGCATATTTTATCA TGCTGGCAGCTCTAGATTATTAACTGTTGGTAATCCATATTTTAGGGTT CCTGCAGGTGGTGGCAATAAGCAGGATATTCCTAAGGTTTCTGCATACC AATATAGAGTATTTAGGGTGCAGTTACCTGACCCAAATAAATTTGGTTT ACCTGATACTAGTATTTATAATCCTGAAACACAACGTTTAGTGTGGGCC TGTGCTGGAGTGGAAATTGGCCGTGGTCAGCCTTTAGGTGTTGGCCTTA GTGGGCATCCATTTTATAATAAATTAGATGACACTGAAAGTTCCCATGC CGCCACGTCTAATGTTTCTGAGGACGTTAGGGACAATGTGTCTGTAGAT TATAAGCAGACACAGTTATGTATTTTGGGCTGTGCCCCTGCTATTGGGG AACACTGGGCTAAAGGCACTGCTTGTAAATCGCGTCCTTTATCACAGGG CGATTGCCCCCCTTTAGAACTTAAAAACACAGTTTTGGAAGATGGTGAT ATGGTAGATACTGGATACGGCGCCATGGACTTCAGCACCCTGCAGGACA CCAAGTGCGAGGTGCCCCTGGACATCTGCCAGAGCATCTGCAAGTACCC CGACTACCTGCAGATGAGCGCCGACCCCTACGGCGACAGCATGTTCTTC TGCCTGAGGAGGGAGCAGCTGTTCGCCAGGCACTTCTGGAACAGGGCCG GCGTGATGGGCGACACCGTGCCCACCGACCTGTACATCAAGGGCACCAG CGCCAACATGAGGGAGACCCCCGGCAGCTGCGTGTACAGCCCCAGCCCC AGCGGCAGCATCGTTACCTCTGACTCCCAGTTGTTTAATAAACCATATT GGTTACATAAGGCACAGGGTCATAACAATGGTGTTTGCTGGCATAATCA ATTATTTGTTACTGTGGTAGATACCACTCGCAGTACCAATTTAACAATA TGTGCTTCTACACAGTCTCCTGTACCTGGGCAATATGATGCTACCAAAT TTAAGCAGTATAGCAGACATGTTGAGGAATATGATTTGCAGTTTATTTT TCAGTTGTGTACTATTACTTTAACTGCAGATGTTATGTCCTATATTCAT AGTATGAATAGCAGTATTTTAGAGGATTGGAACTTTGGTGTTCCCCCCC CGCCAACTACTAGTTTGGTGGATACATATCGTTTTGTACAATCTGTTGC TATTGCCTGTCAAAAGGATGCTGCACCGGCTGAAAATAAGGATCCCTAT GATAAGTTAAAGTTTTGGAATGTGGATTTAAAGGAAAAGTTTTCTTTAG ACTTAGATCAATATCCCCTTGGACGTAAATTTTTGGTTCAGGCTGGATT GCGTCGCAAGCCCACCATAGGCCCTCGCAAACGTTCTGCTCCATCTGCC ACTACGGCTTCTAAACCTGCCAAGCGTGTGCGTGTACGTGCCAGGAAGT AA

(39) TABLE-US-00028 Sequence 27 (SEQ ID NO: 27): ATGCGGCCTAGTGACAATACCGTATATCTTCCACCTCCTTCTGTGGCAA GAGTTGTAAATACCGATGATTACGTGACTCGCACAAGCATATTTTATCA TGCTGGCAGCTCTAGATTATTAACTGTTGGTAATCCATATTTTAGGGTT CCTGCAGGTGGTGGCAATAAGCAGGATATTCCTAAGGTTTCTGCATACC AATATAGAGTATTTAGGGTGCAGTTACCTGACCCAAATAAATTTGGTTT ACCTGATACTAGTATTTATAATCCTGAAACACAACGTTTAGTGTGGGCC TGTGCTGGAGTGGAAATTGGCCGTGGTCAGCCTTTAGGTGTTGGCCTTA GTGGGCATCCATTTTATAATAAATTAGATGACACTGAAAGTTCCCATGC CGCCACGTCTAATGTTTCTGAGGACGTTAGGGACAATGTGTCTGTAGAT TATAAGCAGACACAGTTATGTATTTTGGGCTGTGCCCCTGCTATTGGGG AACACTGGGCTAAAGGCACTGCTTGTAAATCGCGTCCTTTATCACAGGG CGATTGCCCCCCTTTAGAACTTAAAAACACAGTTTTGGAAGATGGTGAT ATGGTAGATACTGGATATGGTGCCATGGACTTTAGTACATTGCAAGATA CTAAATGTGAGGTACCATTGGATATTTGTCAGTCTATTTGTAAATATCC TGATTATTTACAAATGTCTGCAGATCCTTATGGGATTCCATGTTTTTTT GCTTACGGCGTGAGCAGCTTTTTGCTAGGCATTTTTGGAATAGAGCAGG TACTATGGGTGACACTGTGCCTCAATCCTTATATATTAAAGGCACAGGT ATGCGTGCTTCACCTGGCAGCTGTGTGTATTCTCCCTCTCCAAGTGGCT CTATTGTTACCTCTGACTCCCAGTTGTTTAATAAACCATATTGGTTACA TAAGGCACAGGGTCATAACAATGGTGTTTGCTGGCATAATCAATTATTT GTTACTGTGGTAGATACCACTCGCAGTACCAATTTAACAATATGCGCCA GCACCCAGAACCCCGTGCCCAGCACCTACGACCCCACCAAGTTCAAGCA GTACAGCAGGCACGTGGAGGAGTACGACCTGCAGTTCATCTTCCAGCTG TGCACCATCACCCTGACCGCCGATGTTATGTCCTATATTCATAGTATGA ATAGCAGTATTTTAGAGGATTGGAACTTTGGTGTTCCCCCCCCGCCAAC TACTAGTTTGGTGGATACATATCGTTTTGTACAATCTGTTGCTATTGCC TGTCAAAAGGATGCTGCACCGGCTGAAAATAAGGATCCCTATGATAAGT TAAAGTTTTGGAATGTGGATTTAAAGGAAAAGTTTTCTTTAGACTTAGA TCAATATCCCCTTGGACGTAAATTTTTGGTTCAGGCTGGATTGCGTCGC AAGCCCACCATAGGCCCTCGCAAACGTTCTGCTCCATCTGCCACTACGG CTTCTAAACCTGCCAAGCGTGTGCGTGTACGTGCCAGGAAGTAA

(40) TABLE-US-00029 Sequence 28 (SEQ ID NO: 28): ATGCGGCCTAGTGACAATACCGTATATCTTCCACCTCCTTCTGTGGCAA GAGTTGTAAATACCGATGATTACGTGACTCGCACAAGCATATTTTATCA TGCTGGCAGCTCTAGATTATTAACTGTTGGTAATCCATATTTCAAGGTG CCCAAGGGCGGCAACGGCAGGCAGGACGTGCCCAAGGTGAGCGCATACC AATATAGAGTATTTAGGGTGCAGTTACCTGACCCAAATAAATTTGGTTT ACCTGATACTAGTATTTATAATCCTGAAACACAACGTTTAGTGTGGGCC TGTGCTGGAGTGGAAATTGGCCGTGGTCAGCCTTTAGGTGTTGGCCTTA GTGGGCATCCATTTTATAATAAATTAGATGACACTGAAAGTTCCCATGC CGCCACGTCTAATGTTTCTGAGGACGTTAGGGACAATGTGTCTGTAGAT TATAAGCAGACACAGTTATGTATTTTGGGCTGTGCCCCCGCCATCGGCG AGCACTGGGCCAAGGGCACCCTGTGCAAGCCCGCCCAGCTGCAGCCCGG CGACTGCCCCCCCCTGGAGCTGAAGAACACCGTTTTGGAAGATGGTGAT ATGGTAGATACTGGATATGGTGCCATGGACTTTAGTACATTGCAAGATA CTAAATGTGAGGTACCATTGGATATTTGTCAGTCTATTTGTAAATATCC TGATTATTTACAAATGTCTGCAGATCCTTATGGGGATTCCATGTTTTTT TGCTTACGGCGTGAGCAGCTTTTTGCTAGGCATTTTTGGAATAGAGCAG GTACTATGGGTGACACTGTGCCTCAATCCTTATATATTAAAGGCACAGG TATGCGTGCTTCACCTGGCAGCTGTGTGTATTCTCCCTCTCCAAGTGGC TCTATTGTTACCTCTGACTCCCAGTTGTTTAATAAACCATATTGGTTAC ATAAGGCACAGGGTCATAACAATGGTGTTTGCTGGCATAATCAATTATT TGTTACTGTGGTAGATACCACTCGCAGTACCAATTTAACAATATGTGCT TCTACACAGTCTCCTGTACCTGGGCAATATGATGCTACCAAATTTAAGC AGTATAGCAGACATGTTGAGGAATATGATTTGCAGTTTATTTTTCAGTT GTGTACTATTACTTTAACTGCAGATGTTATGTCCTATATTCATAGTATG AATAGCAGTATTTTAGAGGATTGGAACTTTGGTGTTCCCCCCCCGCCAA CTACTAGTTTGGTGGATACATATCGTTTTGTACAATCTGTTGCTATTGC CTGTCAAAAGGATGCTGCACCGGCTGAAAATAAGGATCCCTATGATAAG TTAAAGTTTTGGAATGTGGATTTAAAGGAAAAGTTTTCTTTAGACTTAG ATCAATATCCCCTTGGACGTAAATTTTTGGTTCAGGCTGGATTGCGTCG CAAGCCCACCATAGGCCCTCGCAAACGTTCTGCTCCATCTGCCACTACG GCTTCTAAACCTGCCAAGCGTGTGCGTGTACGTGCCAGGAAGTAA

(41) TABLE-US-00030 Sequence 29 (SEQ ID NO: 29): ATGCGGCCTAGTGACAATACCGTATATCTTCCACCTCCTTCTGTGGCAA GAGTTGTAAATACCGATGATTACGTGACTCGCACAAGCATATTTTATCA TGCTGGCAGCTCTAGATTATTAACTGTTGGTAATCCATATTTTAGGGTT CCTGCAGGTGGTGGCAATAAGCAGGATATTCCTAAGGTTTCTGCATACC AATATAGAGTATTTAGGGTGCAGTTACCTGACCCAAATAAATTTGGTTT ACCTGATACTAGTATTTATAATCCTGAAACACAACGTTTAGTGTGGGCC TGTGCTGGCGTGGAGATCGGCAGGGGCCAGCCCCTGGGCGTGGGCCTGA GCGGCCACCCCCTGTACAACAAGCTGGACGACACCGAGAACAGCCACGT GGCCAGCGCCGTGGACACCAAGGACACCAGGGACAACGTGAGCGTGGAC TACAAGCAGACCCAGCTGTGCATCTTGGGCTGTGCCCCCGCCATCGGCG AGCACTGGGCCAAGGGCACCCTGTGCAAGCCCGCCCAGCTGCAGCCCGG CGACTGCCCCCCCCTGGAGCTGAAGAACACCGTTTTGGAAGATGGTGAT ATGGTAGATACTGGATATGGTGCCATGGACTTTAGTACATTGCAAGATA CTAAATGTGAGGTACCATTGGATATTTGTCAGTCTATTTGTAAATATCC TGATTATTTACAAATGTCTGCAGATCCTTATGGGGATTCCATGTTTTTT TGCTTACGGCGTGAGCAGCTTTTTGCTAGGCATTTTTGGAATAGAGCAG GTACTATGGGTGACACTGTGCCTCAATCCTTATATATTAAAGGCACAGG TATGCGTGCTTCACCTGGCAGCTGTGTGTATTCTCCCTCTCCAAGTGGC TCTATTGTTACCTCTGACTCCCAGTTGTTTAATAAACCATATTGGTTAC ATAAGGCACAGGGTCATAACAATGGTGTTTGCTGGCATAATCAATTATT TGTTACTGTGGTAGATACCACTCGCAGTACCAATTTAACAATATGTGCT TCTACACAGTCTCCTGTACCTGGGCAATATGATGCTACCAAATTTAAGC AGTATAGCAGACATGTTGAGGAATATGATTTGCAGTTTATTTTTCAGTT GTGTACTATTACTTTAACTGCAGATGTTATGTCCTATATTCATAGTATG AATAGCAGTATTTTAGAGGATTGGAACTTTGGTGTTCCCCCCCCGCCAA CTACTAGTTTGGTGGATACATATCGTTTTGTACAATCTGTTGCTATTGC CTGTCAAAAGGATGCTGCACCGGCTGAAAATAAGGATCCCTATGATAAG TTAAAGTTTTGGAATGTGGATTTAAAGGAAAAGTTTTCTTTAGACTTAG ATCAATATCCCCTTGGACGTAAATTTTTGGTTCAGGCTGGATTGCGTCG CAAGCCCACCATAGGCCCTCGCAAACGTTCTGCTCCATCTGCCACTACG GCTTCTAAACCTGCCAAGCGTGTGCGTGTACGTGCCAGGAAGTAA

(42) TABLE-US-00031 Sequence 30 (SEQ ID NO: 30): ATGCGGCCTAGTGACAATACCGTATATCTTCCACCTCCTTCTGTGGCAA GAGTTGTAAATACCGATGATTACGTGACTCGCACAAGCATATTTTATCA TGCTGGCAGCTCTAGATTATTAACTGTTGGTAATCCATATTTTAGGGTT CCTGCAGGTGGTGGCAATAAGCAGGATATTCCTAAGGTTTCTGCATACC AATATAGAGTATTTAGGGTGCAGTTACCTGACCCAAATAAATTTGGTTT ACCTGATACTAGTATTTATAATCCTGAAACACAACGTTTAGTGTGGGCC TGTGCTGGAGTGGAAATTGGCCGTGGTCAGCCTTTAGGTGTTGGCCTTA GTGGGCATCCATTTTATAATAAATTAGATGACACTGAAAGTTCCCATGC CGCCACGTCTAATGTTTCTGAGGACGTTAGGGACAATGTGTCTGTAGAT TATAAGCAGACACAGTTATGTATTTTGGGCTGTGCCCCCGCCATCGGCG AGCACTGGGCCAAGGGCACCCTGTGCAAGCCCGCCCAGCTGCAGCCCGG CGACTGCCCCCCCCTGGAGCTGAAGAACACCGTTTTGGAAGATGGTGAT ATGGTAGATACTGGATATGGTGCCATGGACTTTAGTACATTGCAAGATA CTAAATGTGAGGTACCATTGGATATTTGTCAGTCTATTTGTAAATATCC TGATTATTTACAAATGTCTGCAGATCCTTATGGGGATTCCATGTTTTTT TGCTTACGGCGTGAGCAGCTTTTCGCCAGGCACTTCTGGAACAGGAGCG GCACCATGGGCGACCAGCTGCCCGAGAGCCTGTACATCAAGGGCACCGA CATCAGGGCCAACCCCGGCAGCTACCTGTACAGCCCCAGCCCCAGCGGC TCTATTGTTACCTCTGACTCCCAGTTGTTTAATAAACCATATTGGTTAC ATAAGGCACAGGGTCATAACAATGGTGTTTGCTGGCATAATCAATTATT TGTTACTGTGGTAGATACCACTCGCAGTACCAATTTAACAATATGTGCT TCTACACAGTCTCCTGTACCTGGGCAATATGATGCTACCAAATTTAAGC AGTATAGCAGACATGTTGAGGAATATGATTTGCAGTTTATTTTTCAGTT GTGTACTATTACTTTAACTGCAGATGTTATGTCCTATATTCATAGTATG AATAGCAGTATTTTAGAGGATTGGAACTTTGGTGTTCCCCCCCCGCCAA CTACTAGTTTGGTGGATACATATCGTTTTGTACAATCTGTTGCTATTGC CTGTCAAAAGGATGCTGCACCGGCTGAAAATAAGGATCCCTATGATAAG TTAAAGTTTTGGAATGTGGATTTAAAGGAAAAGTTTTCTTTAGACTTAG ATCAATATCCCCTTGGACGTAAATTTTTGGTTCAGGCTGGATTGCGTCG CAAGCCCACCATAGGCCCTCGCAAACGTTCTGCTCCATCTGCCACTACG GCTTCTAAACCTGCCAAGCGTGTGCGTGTACGTGCCAGGAAGTAA

(43) TABLE-US-00032 Sequence 31 (SEQ ID NO: 31): ATGCGGCCTAGTGACAATACCGTATATCTTCCACCTCCTTCTGTGGCAA GAGTTGTAAATACCGATGATTACGTGACTCGCACAAGCATATTTTATCA TGCTGGCAGCTCTAGATTATTAACTGTTGGTAATCCATATTTTAGGGTT CCTGCAGGTGGTGGCAATAAGCAGGATATTCCTAAGGTTTCTGCATACC AATATAGAGTATTTAGGGTGCAGTTACCTGACCCAAATAAATTTGGTTT ACCTGATACTAGTATTTATAATCCTGAAACACAACGTTTAGTGTGGGCC TGTGCTGGAGTGGAAATTGGCCGTGGTCAGCCTTTAGGTGTTGGCCTTA GTGGGCATCCATTTTATAATAAATTAGATGACACTGAAAGTTCCCATGC CGCCACGTCTAATGTTTCTGAGGACGTTAGGGACAATGTGTCTGTAGAT TATAAGCAGACACAGTTATGTATTTTGGGCTGTGCCCCCGCCATCGGCG AGCACTGGGCCAAGGGCACCCTGTGCAAGCCCGCCCAGCTGCAGCCCGG CGACTGCCCCCCCCTGGAGCTGAAGAACACCGTTTTGGAAGATGGTGAT ATGGTAGATACTGGATATGGTGCCATGGACTTTAGTACATTGCAAGATA CTAAATGTGAGGTACCATTGGATATTTGTCAGTCTATTTGTAAATATCC TGATTATTTACAAATGTCTGCAGATCCTTATGGGGATTCCATGTTTTTT TGCTTACGGCGTGAGCAGCTTTTTGCTAGGCATTTTTGGAATAGAGCAG GTACTATGGGTGACACTGTGCCTCAATCCTTATATATTAAAGGCACAGG TATGCGTGCTTCACCTGGCAGCTGTGTGTATTCTCCCTCTCCAAGTGGC TCTATTGTTACCTCTGACTCCCAGTTGTTTAATAAACCATATTGGTTAC ATAAGGCACAGGGTCATAACAATGGTGTTTGCTGGCATAATCAATTATT TGTTACTGTGGTAGATACCACTCGCAGTACCAATTTAACAATATGCGCC AGCACCACCAGCAGCATCCCCAACGTGTACACCCCCACCAGCTTCAAGC AGTATAGCAGACATGTTGAGGAATATGATTTGCAGTTTATTTTTCAGTT GTGTACTATTACTTTAACTGCAGATGTTATGTCCTATATTCATAGTATG AATAGCAGTATTTTAGAGGATTGGAACTTTGGTGTTCCCCCCCCGCCAA CTACTAGTTTGGTGGATACATATCGTTTTGTACAATCTGTTGCTATTGC CTGTCAAAAGGATGCTGCACCGGCTGAAAATAAGGATCCCTATGATAAG TTAAAGTTTTGGAATGTGGATTTAAAGGAAAAGTTTTCTTTAGACTTAG ATCAATATCCCCTTGGACGTAAATTTTTGGTTCAGGCTGGATTGCGTCG CAAGCCCACCATAGGCCCTCGCAAACGTTCTGCTCCATCTGCCACTACG GCTTCTAAACCTGCCAAGCGTGTGCGTGTACGTGCCAGGAAGTAA

(44) TABLE-US-00033 Sequence 32 (SEQ ID NO: 32): ATGCGGCCTAGTGACAATACCGTATATCTTCCACCTCCTTCTGTGGCAA GAGTTGTAAATACCGATGATTACGTGACTCGCACAAGCATATTTTATCA TGCTGGCAGCTCTAGATTATTAACTGTTGGTAATCCCTACTTCAAGGTG CCCAAGGGCGGCAACGGCAGGCAGGACGTGCCCAAGGTGAGCGCCTACC AGTACAGGGTATTTAGGGTGCAGTTACCTGACCCAAATAAATTTGGTTT ACCTGATACTAGTATTTATAATCCTGAAACACAACGTTTAGTGTGGGCC TGTGCTGGAGTGGAAATTGGCCGTGGTCAGCCTTTAGGTGTTGGCCTTA GTGGGCATCCATTTTATAATAAATTAGATGACACTGAAAGTTCCCATGC CGCCACGTCTAATGTTTCTGAGGACGTTAGGGACAATGTGTCTGTAGAT TATAAGCAGACACAGTTATGTATTTTGGGCTGTGCCCCTGCTATTGGGG AACACTGGGCTAAAGGCACTGCTTGTAAATCGCGTCCTTTATCACAGGG CGATTGCCCCCCTTTAGAACTTAAAAACACAGTTTTGGAAGATGGTGAT ATGGTAGATACTGGATACGGCGCCATGGACTTCAGCACCCTGCAGGACA CCAAGTGCGAGGTGCCCCTGGACATCTGCCAGAGCATCTGCAAGTACCC CGACTACCTGCAGATGAGCGCCGACCCCTACGGCGACAGCATGTTCTTC TGCCTGAGGAGGGAGCAGCTGTTCGCCAGGCACTTCTGGAACAGGGCCG GCGTGATGGGCGACACCGTGCCCACCGACCTGTACATCAAGGGCACCAG CGCCAACATGAGGGAGACCCCCGGCAGCTGCGTGTACAGCCCCAGCCCC AGCGGCAGCATCGTTACCTCTGACTCCCAGTTGTTTAATAAACCATATT GGTTACATAAGGCACAGGGTCATAACAATGGTGTTTGCTGGCATAATCA ATTATTTGTTACTGTGGTAGATACCACTCGCAGTACCAATTTAACAATA TGTGCTTCTACACAGTCTCCTGTACCTGGGCAATATGATGCTACCAAAT TTAAGCAGTATAGCAGACATGTTGAGGAATATGATTTGCAGTTTATTTT TCAGTTGTGTACTATTACTTTAACTGCAGATGTTATGTCCTATATTCAT AGTATGAATAGCAGTATTTTAGAGGATTGGAACTTTGGTGTTCCCCCCC CGCCAACTACTAGTTTGGTGGATACATATCGTTTTGTACAATCTGTTGC TATTGCCTGTCAAAAGGATGCTGCACCGGCTGAAAATAAGGATCCCTAT GATAAGTTAAAGTTTTGGAATGTGGATTTAAAGGAAAAGTTTTCTTTAG ACTTAGATCAATATCCCCTTGGACGTAAATTTTTGGTTCAGGCTGGATT GCGTCGCAAGCCCACCATAGGCCCTCGCAAACGTTCTGCTCCATCTGCC ACTACGGCTTCTAAACCTGCCAAGCGTGTGCGTGTACGTGCCAGGAAGT AA

(45) TABLE-US-00034 Sequence 33 (SEQ ID NO: 33): ATGCGGCCTAGTGACAATACCGTATATCTTCCACCTCCTTCTGTGGCAA GAGTTGTAAATACCGATGATTACGTGACTCGCACAAGCATATTTTATCA TGCTGGCAGCTCTAGATTATTAACTGTTGGTAATCCATATTTTAGGGTT CCTGCAGGTGGTGGCAATAAGCAGGATATTCCTAAGGTTTCTGCATACC AATATAGAGTATTTAGGGTGCAGTTACCTGACCCAAATAAATTTGGTTT ACCTGATACTAGTATTTATAATCCTGAAACACAACGTTTAGTGTGGGCC TGTGCTGGCGTGGAGATCGGCAGGGGCCAGCCCCTGGGCGTGGGCCTGA GCGGCCACCCCCTGTACAACAAGCTGGACGACACCGAGAACAGCCACGT GGCCAGCGCCGTGGACACCAAGGACACCAGGGACAACGTGAGCGTGGAC TACAAGCAGACCCAGCTGTGCATCTTGGGCTGTGCCCCTGCTATTGGGG AACACTGGGCTAAAGGCACTGCTTGTAAATCGCGTCCTTTATCACAGGG CGATTGCCCCCCTTTAGAACTTAAAAACACAGTTTTGGAAGATGGTGAT ATGGTAGATACTGGATACGGCGCCATGGACTTCAGCACCCTGCAGGACA CCAAGTGCGAGGTGCCCCTGGACATCTGCCAGAGCATCTGCAAGTACCC CGACTACCTGCAGATGAGCGCCGAcCCCTACGGCGACAGCATGTTCTTC TGCCTGAGGAGGGAGCAGCTGTTCGCCAGGCACTTCTGGAACAGGGCCG GCGTGATGGGCGACACCGTGCCCACCGACCTGTACATCAAGGGCACCAG CGCCAACATGAGGGAGACCCCCGGCAGCTGCGTGTACAGCCCCAGCCCC AGCGGCAGCATCGTTACCTCTGACTCCCAGTTGTTTAATAAACCATATT GGTTACATAAGGCACAGGGTCATAACAATGGTGTTTGCTGGCATAATCA ATTATTTGTTACTGTGGTAGATACCACTCGCAGTACCAATTTAACAATA TGTGCTTCTACACAGTCTCCTGTACCTGGGCAATATGATGCTACCAAAT TTAAGCAGTATAGCAGACATGTTGAGGAATATGATTTGCAGTTTATTTT TCAGTTGTGTACTATTACTTTAACTGCAGATGTTATGTCCTATATTCAT AGTATGAATAGCAGTATTTTAGAGGATTGGAACTTTGGTGTTCCCCCCC CGCCAACTACTAGTTTGGTGGATACATATCGTTTTGTACAATCTGTTGC TATTGCCTGTCAAAAGGATGCTGCACCGGCTGAAAATAAGGATCCCTAT GATAAGTTAAAGTTTTGGAATGTGGATTTAAAGGAAAAGTTTTCTTTAG ACTTAGATCAATATCCCCTTGGACGTAAATTTTTGGTTCAGGCTGGATT GCGTCGCAAGCCCACCATAGGCCCTCGCAAACGTTCTGCTCCATCTGCC ACTACGGCTTCTAAACCTGCCAAGCGTGTGCGTGTACGTGCCAGGAAGT AA

(46) TABLE-US-00035 Sequence 34 (SEQ ID NO: 34): ATGCGGCCTAGTGACAATACCGTATATCTTCCACCTCCTTCTGTGGCAA GAGTTGTAAATACCGATGATTACGTGACTCGCACAAGCATATTTTATCA TGCTGGCAGCTCTAGATTATTAACTGTTGGTAATCCATATTTTAGGGTT CCTGCAGGTGGTGGCAATAAGCAGGATATTCCTAAGGTTTCTGCATACC AATATAGAGTATTTAGGGTGCAGTTACCTGACCCAAATAAATTTGGTTT ACCTGATACTAGTATTTATAATCCTGAAACACAACGTTTAGTGTGGGCC TGTGCTGGAGTGGAAATTGGCCGTGGTCAGCCTTTAGGTGTTGGCCTTA GTGGGCATCCATTTTATAATAAATTAGATGACACTGAAAGTTCCCATGC CGCCACGTCTAATGTTTCTGAGGACGTTAGGGACAATGTGTCTGTAGAT TATAAGCAGACACAGTTATGTATTTTGGGCTGTGCCCCCGCCATCGGCG AGCACTGGACCAAGGGCACCGCCTGCAAGCCCACCACCGTGGTGCAGGG CGACTGCCCCCCCCTGGAGCTGAAAAACACAGTTTTGGAAGATGGTGAT ATGGTAGATACTGGATACGGCGCCATGGACTTCAGCACCCTGCAGGACA CCAAGTGCGAGGTGCCCCTGGACATCTGCCAGAGCATCTGCAAGTACCC CGACTACCTGCAGATGAGCGCCGACCCCTACGGCGACAGCATGTTCTTC TGCCTGAGGAGGGAGCAGCTGTTCGCCAGGCACTTCTGGAACAGGGCCG GCGTGATGGGCGACACCGTGCCCACCGACCTGTACATCAAGGGCACCAG CGCCAACATGAGGGAGACCCCCGGCAGCTGCGTGTACAGCCCCAGCCCC AGCGGCAGCATCGTTACCTCTGACTCCCAGTTGTTTAATAAACCATATT GGTTACATAAGGCACAGGGTCATAACAATGGTGTTTGCTGGCATAATCA ATTATTTGTTACTGTGGTAGATACCACTCGCAGTACCAATTTAACAATA TGTGCTTCTACACAGTCTCCTGTACCTGGGCAATATGATGCTACCAAAT TTAAGCAGTATAGCAGACATGTTGAGGAATATGATTTGCAGTTTATTTT TCAGTTGTGTACTATTACTTTAACTGCAGATGTTATGTCCTATATTCAT AGTATGAATAGCAGTATTTTAGAGGATTGGAACTTTGGTGTTCCCCCCC CGCCAACTACTAGTTTGGTGGATACATATCGTTTTGTACAATCTGTTGC TATTGCCTGTCAAAAGGATGCTGCACCGGCTGAAAATAAGGATCCCTAT GATAAGTTAAAGTTTTGGAATGTGGATTTAAAGGAAAAGTTTTCTTTAG ACTTAGATCAATATCCCCTTGGACGTAAATTTTTGGTTCAGGCTGGATT GCGTCGCAAGCCCACCATAGGCCCTCGCAAACGTTCTGCTCCATCTGCC ACTACGGCTTCTAAACCTGCCAAGCGTGTGCGTGTACGTGCCAGGAAGT AA

(47) TABLE-US-00036 Sequence 35 (SEQ ID NO: 35): ATGCGGCCTAGTGACAATACCGTATATCTTCCACCTCCTTCTGTGGCAA GAGTTGTAAATACCGATGATTACGTGACTCGCACAAGCATATTTTATCA TGCTGGCAGCTCTAGATTATTAACTGTTGGTAATCCATATTTTAGGGTT CCTGCAGGTGGTGGCAATAAGCAGGATATTCCTAAGGTTTCTGCATACC AATATAGAGTATTTAGGGTGCAGTTACCTGACCCAAATAAATTTGGTTT ACCTGATACTAGTATTTATAATCCTGAAACACAACGTTTAGTGTGGGCC TGTGCTGGAGTGGAAATTGGCCGTGGTCAGCCTTTAGGTGTTGGCCTTA GTGGGCATCCATTTTATAATAAATTAGATGACACTGAAAGTTCCCATGC CGCCACGTCTAATGTTTCTGAGGACGTTAGGGACAATGTGTCTGTAGAT TATAAGCAGACACAGTTATGTATTTTGGGCTGTGCCCCTGCTATTGGGG AACACTGGGCTAAAGGCACTGCTTGTAAATCGCGTCCTTTATCACAGGG CGATTGCCCCCCTTTAGAACTTAAAAACACAGTTTTGGAAGATGGTGAT ATGGTAGATACTGGATACGGCGCCATGGACTTCAGCACCCTGCAGGACA CCAAGTGCGAGGTGCCCCTGGACATCTGCCAGAGCATCTGCAAGTACCC CGACTACCTGCAGATGAGCGCCGAcCCCTACGGCGACAGCATGTTCTTC TGCCTGAGGAGGGAGCAGCTGTTCGCCAGGCACTTCTGGAACAGGGCCG GCGTGATGGGCGACACCGTGCCCACCGACCTGTACATCAAGGGCACCAG CGCCAACATGAGGGAGACCCCCGGCAGCTGCGTGTACAGCCCCAGCCCC AGCGGCAGCATCGTTACCTCTGACTCCCAGTTGTTTAATAAACCATATT GGTTACATAAGGCACAGGGTCATAACAATGGTGTTTGCTGGCATAATCA ATTATTTGTTACTGTGGTAGATACCACTCGCAGTACCAATTTAACAATA TGCGCCAGCACCACCAGCAGCATCCCCAACGTGTACACCCCCACCAGCT TCAAGCAGTATAGCAGACATGTTGAGGAATATGATTTGCAGTTTATTTT TCAGTTGTGTACTATTACTTTAACTGCAGATGTTATGTCCTATATTCAT AGTATGAATAGCAGTATTTTAGAGGATTGGAACTTTGGTGTTCCCCCCC CGCCAACTACTAGTTTGGTGGATACATATCGTTTTGTACAATCTGTTGC TATTGCCTGTCAAAAGGATGCTGCACCGGCTGAAAATAAGGATCCCTAT GATAAGTTAAAGTTTTGGAATGTGGATTTAAAGGAAAAGTTTTCTTTAG ACTTAGATCAATATCCCCTTGGACGTAAATTTTTGGTTCAGGCTGGATT GCGTCGCAAGCCCACCATAGGCCCTCGCAAACGTTCTGCTCCATCTGCC ACTACGGCTTCTAAACCTGCCAAGCGTGTGCGTGTACGTGCCAGGAAGT AA

(48) TABLE-US-00037 Sequence 36 (SEQ ID NO: 36): ATGCGGCCTAGTGACAATACCGTATATCTTCCACCTCCTTCTGTGGCAA GAGTTGTAAATACCGATGATTACGTGACTAGGACCAGCATCTTCTACCA CGCCGGCAGCAGCAGGCTGCTGACCGTGGGCAACCCCTACTTCAGGGTG GTGCCCAACGGCGCCGGCAACAAGCAGGCCGTGCCCAAGGTGAGCGCCT ACCAGTACAGGGTGTTCAGGGTGCAGTTACCTGACCCAAATAAATTTGG TTTACCTGATACTAGTATTTATAATCCTGAAACACAACGTTTAGTGTGG GCCTGTGCTGGAGTGGAAATTGGCCGTGGTCAGCCTTTAGGTGTTGGCC TTAGTGGGCATCCATTTTATAATAAATTAGATGACACTGAAAGTTCCCA TGCCGCCACGTCTAATGTTTCTGAGGACGTTAGGGACAATGTGTCTGTA GATTATAAGCAGACACAGTTATGTATTTTGGGCTGTGCCCCTGCTATTG GGGAACACTGGGCTAAAGGCACTGCTTGTAAATCGCGTCCTTTATCACA GGGCGATTGCCCCCCTTTAGAACTTAAAAACACAGTTTTGGAAGATGGT GATATGGTAGATACTGGATATGGTGCCATGGACTTTAGTACATTGCAAG ATACTAAATGTGAGGTACCATTGGATATTTGTCAGTCTATTTGTAAATA TCCTGATTATTTACAAATGTCTGCAGATCCTTATGGGGATTCCATGTTT TTTTGCTTACGGCGTGAGCAGCTTTTTGCTAGGCATTTTTGGAATAGAG CAGGTACTATGGGTGACACTGTGCCTCAATCCTTATATATTAAAGGCAC AGGTATGCGTGCTTCACCTGGCAGCTGTGTGTATTCTCCCTCTCCAAGT GGCTCTATTGTTACCTCTGACTCCCAGTTGTTTAATAAACCATATTGGT TACATAAGGCACAGGGTCATAACAATGGTGTTTGCTGGCATAATCAATT ATTTGTTACTGTGGTAGATACCACTCGCAGTACCAATTTAACAATATGC GCCAGCACCACCAGCAGCATCCCCAACGTGTACACCCCCACCAGCTTCA AGCAGTATAGCAGACATGTTGAGGAATATGATTTGCAGTTTATTTTTCA GTTGTGTACTATTACTTTAACTGCAGATGTTATGTCCTATATTCATAGT ATGAATAGCAGTATTTTAGAGGATTGGAACTTTGGTGTTCCCCCCCCGC CAACTACTAGTTTGGTGGATACATATCGTTTTGTACAATCTGTTGCTAT TGCCTGTCAAAAGGATGCTGCACCGGCTGAAAATAAGGATCCCTATGAT AAGTTAAAGTTTTGGAATGTGGATTTAAAGGAAAAGTTTTCTTTAGACT TAGATCAATATCCCCTTGGACGTAAATTTTTGGTTCAGGCTGGATTGCG TCGCAAGCCCACCATAGGCCCTCGCAAACGTTCTGCTCCATCTGCCACT ACGGCTTCTAAACCTGCCAAGCGTGTGCGTGTACGTGCCAGGAAGTAA

(49) TABLE-US-00038 Sequence 37 (SEQ ID NO: 37): ATGCGGCCTAGTGACAATACCGTATATCTTCCACCTCCTTCTGTGGCAA GAGTTGTAAATACCGATGATTACGTGACTCGCACAAGCATATTTTATCA TGCTGGCAGCTCTAGATTATTAACTGTTGGTAATCCATATTTTAGGGTT CCTGCAGGTGGTGGCAATAAGCAGGATATTCCTAAGGTTTCTGCATACC AATATAGAGTATTTAGGGTGCAGTTACCTGACCCAAATAAATTTGGTTT ACCTGATACTAGTATTTATAATCCTGAAACACAACGTTTAGTGTGGGCC TGTGCTGGAGTGGAGATCGGCAGGGGCCAGCCCCTGGGCATCGGCCTGA GCGGCCACCCCTTCTACAACAAGCTGGACGACACCGAGAGCGCCCACGC CGCCACCGCCGTGATCACCCAGGACGTGAGGGACAACGTGAGCGTGGAC TACAAGCAGACCCAGCTGTGCATCCTGGGCTGCGCCCCTGCTATTGGGG AACACTGGGCTAAAGGCACTGCTTGTAAATCGCGTCCTTTATCACAGGG CGATTGCCCCCCTTTAGAACTTAAAAACACAGTTTTGGAAGATGGTGAT ATGGTAGATACTGGATATGGTGCCATGGACTTTAGTACATTGCAAGATA CTAAATGTGAGGTACCATTGGATATTTGTCAGTCTATTTGTAAATATCC TGATTATTTACAAATGTCTGCAGATCCTTATGGGGATTCCATGTTTTTT TGCTTACGGCGTGAGCAGCTTTTTGCTAGGCATTTTTGGAATAGAGCAG GTACTATGGGTGACACTGTGCCTCAATCCTTATATATTAAAGGCACAGG TATGCGTGCTTCACCTGGCAGCTGTGTGTATTCTCCCTCTCCAAGTGGC TCTATTGTTACCTCTGACTCCCAGTTGTTTAATAAACCATATTGGTTAC ATAAGGCACAGGGTCATAACAATGGTGTTTGCTGGCATAATCAATTATT TGTTACTGTGGTAGATACCACTCGCAGTACCAATTTAACAATATGCGCC AGCACCACCAGCAGCATCCCCAACGTGTACACCCCCACCAGCTTCAAGC AGTATAGCAGACATGTTGAGGAATATGATTTGCAGTTTATTTTTCAGTT GTGTACTATTACTTTAACTGCAGATGTTATGTCCTATATTCATAGTATG AATAGCAGTATTTTAGAGGATTGGAACTTTGGTGTTCCCCCCCCGCCAA CTACTAGTTTGGTGGATACATATCGTTTTGTACAATCTGTTGCTATTGC CTGTCAAAAGGATGCTGCACCGGCTGAAAATAAGGATCCCTATGATAAG TTAAAGTTTTGGAATGTGGATTTAAAGGAAAAGTTTTCTTTAGACTTAG ATCAATATCCCCTTGGACGTAAATTTTTGGTTCAGGCTGGATTGCGTCG CAAGCCCACCATAGGCCCTCGCAAACGTTCTGCTCCATCTGCCACTACG GCTTCTAAACCTGCCAAGCGTGTGCGTGTACGTGCCAGGAAGTAA

(50) TABLE-US-00039 Sequence 38 (SEQ ID NO: 38): ATGCGGCCTAGTGACAATACCGTATATCTTCCACCTCCTTCTGTGGCAA GAGTTGTAAATACCGATGATTACGTGACTAGGACCAGCATCTTCTACCA CGCCGGCAGCAGCAGGCTGCTGACCGTGGGCAACCCCTACTTCAGGGTG GTGCCCAACGGCGCCGGCAACAAGCAGGCCGTGCCCAAGGTGAGCGCCT ACCAGTACAGGGTGTTCAGGGTGCAGTTACCTGACCCAAATAAATTTGG TTTACCTGATACTAGTATTTATAATCCTGAAACACAACGTTTAGTGTGG GCCTGTGCTGGAGTGGAAATTGGCCGTGGTCAGCCTTTAGGTGTTGGCC TTAGTGGGCATCCATTTTATAATAAATTAGATGACACTGAAAGTTCCCA TGCCGCCACGTCTAATGTTTCTGAGGACGTTAGGGACAATGTGTCTGTA GATTATAAGCAGACACAGTTATGTATTTTGGGCTGTGCCCCCGCCATCG GCGAGCACTGGGCCAAGGGCACCCTGTGCAAGCCCGCCCAGCTGCAGCC CGGCGACTGCCCCCCCCTGGAGCTGAAGAACACCGTTTTGGAAGATGGT GATATGGTAGATACTGGATATGGTGCCATGGACTTTAGTACATTGCAAG ATACTAAATGTGAGGTACCATTGGATATTTGTCAGTCTATTTGTAAATA TCCTGATTATTTACAAATGTCTGCAGATCCTTATGGGGATTCCATGTTT TTTTGCTTACGGCGTGAGCAGCTTTTTGCTAGGCATTTTTGGAATAGAG CAGGTACTATGGGTGACACTGTGCCTCAATCCTTATATATTAAAGGCAC AGGTATGCGTGCTTCACCTGGCAGCTGTGTGTATTCTCCCTCTCCAAGT GGCTCTATTGTTACCTCTGACTCCCAGTTGTTTAATAAACCATATTGGT TACATAAGGCACAGGGTCATAACAATGGTGTTTGCTGGCATAATCAATT ATTTGTTACTGTGGTAGATACCACTCGCAGTACCAATTTAACAATATGC GCCAGCACCACCAGCAGCATCCCCAACGTGTACACCCCCACCAGCTTCA AGCAGTATAGCAGACATGTTGAGGAATATGATTTGCAGTTTATTTTTCA GTTGTGTACTATTACTTTAACTGCAGATGTTATGTCCTATATTCATAGT ATGAATAGCAGTATTTTAGAGGATTGGAACTTTGGTGTTCCCCCCCCGC CAACTACTAGTTTGGTGGATACATATCGTTTTGTACAATCTGTTGCTAT TGCCTGTCAAAAGGATGCTGCACCGGCTGAAAATAAGGATCCCTATGAT AAGTTAAAGTTTTGGAATGTGGATTTAAAGGAAAAGTTTTCTTTAGACT TAGATCAATATCCCCTTGGACGTAAATTTTTGGTTCAGGCTGGATTGCG TCGCAAGCCCACCATAGGCCCTCGCAAACGTTCTGCTCCATCTGCCACT ACGGCTTCTAAACCTGCCAAGCGTGTGCGTGTACGTGCCAGGAAGTAA

(51) TABLE-US-00040 Sequence 39 (SEQ ID NO: 39) VPNGAGNKQAV

(52) TABLE-US-00041 Sequence 40 (SEQ ID NO: 40): IGLSGHPFYNKLDDTESAHAATAVITQ

(53) TABLE-US-00042 Sequence 41 (SEQ ID NO: 41) LCKPAQLQP

(54) TABLE-US-00043 Sequence 42 (SEQ ID NO: 42): VMGDTVPTDLYIKGTSANMRET

(55) TABLE-US-00044 Sequence 43 (SEQ ID NO: 43) KVPKGGNGRQDV

(56) TABLE-US-00045 Sequence 44 (SEQ ID NO: 44) TSSIPNVYTPTS

(57) TABLE-US-00046 Sequence 109 (SEQ ID NO: 109) NPVPSTYDP

(58) TABLE-US-00047 Sequence 110 (SEQ ID NO: 110): LYNKLDDTENSHVASAVDTKDT

(59) TABLE-US-00048 Sequence 111 (SEQ ID NO: 111): SGTMGDQLPESLYIKGTDIRANPGSYL

(60) TABLE-US-00049 Sequence 112 (SEQ ID NO: 112) TKGTACKPTTVV

(61) TABLE-US-00050 Sequence 113 (SEQ ID NO: 113): MRPSDNTVYLPPPSVARVVNTDDYVTRTSIFYHAGSSRLLTVGNPYFRV PAGGGNKQDIPKVSAYQYRVFRVQLPDPNKFGLPDTSIYNPETQRLVWA CAGVEIGRGQPLGVGLSGHPFYNKLDDTESSHAATSNVSEDVRDNVSVD YKQTQLCILGCAPAIGEHWAKGTACKSRPLSQGDCPPLELKNTVLEDGD MVDTGYGAMDFSTLQDTKCEVPLDICQSICKYPDYLQMSADPYGDSMFF CLRREQLFARHFWNRAGTMGDTVPQSLYIKGTGMRASPGSCVYSPSPSG SIVTSDSQLFNKPYWLHKAQGHNNGVCWHNQLFVTVVDTTRSTNLTICA STQSPVPGQYDATKFKQYSRHVEEYDLQFIFQLCTITLTADVMSYIHSM NSSILEDWNFGVPPPPTTSLVDTYRFVQSVAIACQKDAAPAENKDPYDK LKFWNVDLKEKFSLDLDQYPLGRKFLVQAGLRRKPTIGPRKRSAPSATT ASKPAKRVRVRARK

(62) TABLE-US-00051 Sequence 114 (SEQ ID NO: 114): ATGCGGCCTAGTGACAATACCGTATATCTTCCACCTCCTTCTGTGGCAA GAGTTGTAAATACCGATGATTACGTGACTCGCACAAGCATATTTTATCA TGCTGGCAGCTCTAGATTATTAACTGTTGGTAATCCATATTTTAGGGTT CCTGCAGGTGGTGGCAATAAGCAGGATATTCCTAAGGTTTCTGCATACC AATATAGAGTATTTAGGGTGCAGTTACCTGACCCAAATAAATTTGGTTT ACCTGATACTAGTATTTATAATCCTGAAACACAACGTTTAGTGTGGGCC TGTGCTGGAGTGGAAATTGGCCGTGGTCAGCCTTTAGGTGTTGGCCTTA GTGGGCATCCATTTTATAATAAATTAGATGACACTGAAAGTTCCCATGC CGCCACGTCTAATGTTTCTGAGGACGTTAGGGACAATGTGTCTGTAGAT TATAAGCAGACACAGTTATGTATTTTGGGCTGTGCCCCTGCTATTGGGG AACACTGGGCTAAAGGCACTGCTTGTAAATCGCGTCCTTTATCACAGGG CGATTGCCCCCCTTTAGAACTTAAAAACACAGTTTTGGAAGATGGTGAT ATGGTAGATACTGGATATGGTGCCATGGACTTTAGTACATTGCAAGATA CTAAATGTGAGGTACCATTGGATATTTGTCAGTCTATTTGTAAATATCC TGATTATTTACAAATGTCTGCAGATCCTTATGGGGATTCCATGTTTTTT TGCTTACGGCGTGAGCAGCTTTTTGCTAGGCATTTTTGGAATAGAGCAG GTACTATGGGTGACACTGTGCCTCAATCcTTATATATTAAAGGCACAGG TATGCGTGCTTCACCTGGCAGCTGTGTGTATTCTCCCTCTCCAAGTGGC TCTATTGTTACCTCTGACTCCCAGTTGTTTAATAAACCATATTGGTTAC ATAAGGCACAGGGTCATAACAATGGTGTTTGCTGGCATAATCAATTATT TGTTACTGTGGTAGATACCACTCGCAGTACCAATTTAACAATATGTGCT TCTACACAGTCTCCTGTACCTGGGCAATATGATGCTACCAAATTTAAGC AGTATAGCAGACATGTTGAGGAATATGATTTGCAGTTTATTTTTCAGTT GTGTACTATTACTTTAACTGCAGATGTTATGTCCTATATTCATAGTATG AATAGCAGTATTTTAGAGGATTGGAACTTTGGTGTTCCCCCCCCGCCAA CTACTAGTTTGGTGGATACATATCGTTTTGTACAATCTGTTGCTATTGC CTGTCAAAAGGATGCTGCACCGGCTGAAAATAAGGATCCCTATGATAAG TTAAAGTTTTGGAATGTGGATTTAAAGGAAAAGTTTTCTTTAGACTTAG ATCAATATCCCCTTGGACGTAAATTTTTGGTTCAGGCTGGATTGCGTCG CAAGCCCACCATAGGCCCTCGCAAACGTTCTGCTCCATCTGCCACTACG GCTTCTAAACCTGCCAAGCGTGTGCGTGTACGTGCCAGGAAGTAA

(63) TABLE-US-00052 Sequence 115 (SEQ ID NO: 115): MALWRPSDSTVYLPPPSVARVVSTDDYVSRTSIFYHAGSSRLLTVGNPY FRVVPNGAGNKQAVPKVSAYQYRVFRVALPDPNKFGLPDSTIYNPETQR LVWACVGMEIGRGQPLGIGLSGHPFYNKLDDTESAHAATAVITQDVRDN VSVDYKQTQLCILGCVPAIGEHWAKGTLCKPAQLQPGDCPPLELKNTII EDGDMVDTGYGAMDFSTLQDTKCEVPLDICQSICKYPDYLQMSADPYGD SMFFCLRREQLFARHFWNRAGVMGDTVPTDLYIKGTSANMRETPGSCVY SPSPSGSIITSDSQLFNKPYWLHKAQGHNNGICWHNQLFVTVVDTTRST NLTLCASTQNPVPSTYDPTKFKQYSRHVEEYDLQFIFQLCTITLTAEVM SYIHSMNSSILENWNFGVPPPPTTSLVDTYRFVQSVAVTCQKDTTPPEK QDPYDKLKFWTVDLKEKFSSDLDQYPLGRKFLVQAGLRRRPTIGPRKRP AASTSTASTASRPAKRVRIRSKK

(64) TABLE-US-00053 Sequence 116 (SEQ ID NO: 116): ATGGCCCTGTGGAGGCCCAGCGACAGCACCGTGTACCTGCCCCCCCCCA GCGTGGCCAGGGTGGTGAGCACCGACGACTACGTGAGCAGGACCAGCAT CTTCTACCACGCCGGCAGCAGCAGGCTGCTGACCGTGGGCAACCCCTAC TTCAGGGTGGTGCCCAACGGCGCCGGCAACAAGCAGGCCGTGCCCAAGG TGAGCGCCTACCAGTACAGGGTGTTCAGGGTGGCCCTGCCCGACCCCAA CAAGTTCGGCCTGCCCGACAGCACCATCTACAACCCCGAGACCCAGAGG CTGGTGTGGGCCTGCGTGGGCATGGAGATCGGCAGGGGCCAGCCCCTGG GCATCGGCCTGAGCGGCCACCCCTTCTACAACAAGCTGGACGACACCGA GAGCGCCCACGCCGCCACCGCCGTGATCACCCAGGACGTGAGGGACAAC GTGAGCGTGGACTACAAGCAGACCCAGCTGTGCATCCTGGGCTGCGTGC CCGCCATCGGCGAGCACTGGGCCAAGGGCACCCTGTGCAAGCCCGCCCA GCTGCAGCCCGGCGACTGCCCCCCCCTGGAGCTGAAGAACACCATCATC GAGGACGGCGACATGGTGGACACCGGCTACGGCGCCATGGACTTCAGCA CCCTGCAGGACACCAAGTGCGAGGTGCCCCTGGACATCTGCCAGAGCAT CTGCAAGTACCCCGACTACCTGCAGATGAGCGCCGACCCCTACGGCGAC AGCATGTTCTTCTGCCTGAGGAGGGAGCAGCTGTTCGCCAGGCACTTCT GGAACAGGGCCGGCGTGATGGGCGACACCGTGCCCACCGACCTGTACAT CAAGGGCACCAGCGCCAACATGAGGGAGACCCCCGGCAGCTGCGTGTAC AGCCCCAGCCCCAGCGGCAGCATCATCACCAGCGACAGCCAGCTGTTCA ACAAGCCCTACTGGCTGCACAAGGCCCAGGGCCACAACAACGGCATCTG CTGGCACAACCAGCTGTTCGTGACCGTGGTGGACACCACCAGGAGCACC AACCTGACCCTGTGCGCCAGCACCCAGAACCCCGTGCCCAGCACCTACG ACCCCACCAAGTTCAAGCAGTACAGCAGGCACGTGGAGGAGTACGACCT GCAGTTCATCTTCCAGCTGTGCACCATCACCCTGACCGCCGAGGTGATG AGCTACATCCACAGCATGAACAGCAGCATCCTGGAGAACTGGAACTTCG GCGTGCCCCCCCCCCCCACCACCAGCCTGGTGGACACCTACAGGTTCGT GCAGAGCGTGGCCGTGACCTGCCAGAAGGACACCACCCCCCCCGAGAAG CAGGACCCCTACGACAAGCTGAAGTTCTGGACCGTGGACCTGAAGGAGA AGTTCAGCAGCGACCTGGACCAGTACCCCCTGGGCAGGAAGTTCCTGGT GCAGGCCGGCCTGAGGAGGAGGCCCACCATCGGCCCCAGGAAGAGGCCC GCCGCCAGCACCAGCACCGCCAGCACCGCCAGCAGGCCCGCCAAGAGGG TGAGGATCAGGAGCAAGAAGTGA

(65) Specific Modes for Carrying Out the Invention

(66) The present invention is further described by reference to the examples as follows, wherein the examples are used only for the purpose of illustrating the present invention, rather than limiting the present invention.

(67) Unless indicated otherwise, the molecular biological experimental methods and immunological assays used in the present invention are carried out substantially in accordance with the methods as described in J. Sambrook et al., Molecular Cloning: A Laboratory Manual (Second Edition), Cold Spring Harbor Laboratory Press, 1989, and F. M. Ausubel et al., Short Protocols in Molecular Biology, 3rd Edition, John Wiley & Sons, Inc., 1995; and restriction enzymes are used under the conditions recommended by the manufacturers. Those skilled in the art understand that the examples are used for illustrating the present invention, but not intended to limit the protection scope of the present invention.

Example 1. Expression and Purification of the Mutated HPV18 L1 Proteins

(68) Construction of Expression Vectors

(69) Gibson assembly (Gibson D G, Young L, Chuang R Y, Venter J C, Hutchison C A, Smith H O. Enzymatic assembly of DNA molecules up to several hundred kilobases. Nat Methods. 2009; 6:343-5. doi: 10.1038/nmeth.1318) was used to construct the expression vector encoding the mutated HPV18 L1 protein comprising a specific segment from HPV45 L1 protein and/or a specific segment from HPV59 L1 protein. In brief, a short fragment comprising mutations and a long fragment comprising no mutation were first obtained by PCR, and Gibson assembly system was then used to ligate the two fragments to form a ring.

(70) The initial template used comprised the plasmid pTO-T7-HPV18N65L1 (encoding the HPV18 L1 protein having 65 amino acids truncated at N-terminal, and the protein was designated as HPV18N65; abbreviated as 18L1N65 in Table 2), the plasmid pTO-T7-HPV45L1N27C (encoding the HPV45 L1 protein having 27 amino acids truncated at N-terminal, and the protein was designated as HPV45N27; abbreviated as 45L1N27 in Table 2), the plasmid pTO-T7-H18N65-45T1 (encoding the mutated protein H18N65-45T1; abbreviated as H18N65-45T1 in Table 2), the plasmid pTO-T7-H18N65-45T2 (encoding the mutated protein H18N65-45T2; abbreviated as H18N65-45T2 in Table 2), the plasmid pTO-T7-H18N65-45T3 (encoding the mutated protein H18N65-45T3; abbreviated as H18N65-45T3 in Table 2), the plasmid pTO-T7-H18N65-45T4 (encoding the mutated protein H18N65-45T4; abbreviated as H18N65-45T4 in Table 2), the plasmid pTO-T7-H18N65-45T3-59S5 (encoding the mutated protein H18N65-45T3-59S5; abbreviated as H18N65-45T3-59S5 in Table 2) and the plasmid pTO-T7-HPV59L1 (encoding the HPV59 L1 protein; abbreviated as 59L1 in Table 2). The templates and primers for each PCR were shown in Table 2, and the amplification conditions for PCR for amplifying the short fragment were as followed: denaturation at 94° C. for 10 min; 25 cycles (denaturation at 94° C. for 50 sec, annealing at a given temperature for a certain period of time, and extension at 72° C. for 1 min); and final extension at 72° C. for 10 min. The amplification conditions for PCR for amplifying the long fragment were as followed: denaturation at 94° C. for 10 min; 25 cycles (denaturation at 94° C. for 50 sec, annealing at a given temperature for a certain period of time, and extension at 72° C. for 7.5 min); and final extension at 72° C. for 10 min. The temperature and time of annealing were listed in Table 2. The sequences of the PCR primers used were listed in Table 3.

(71) The amplification product was subjected to electrophoresis, the fragment of interest was then recovered by using DNA Extraction Kit (BEYOTIME, Cat. No. D0033), and its concentration was determined. The short fragment and long fragment obtained by amplification were mixed at a molar ratio of 2:1 (a total volume of 3 μL), and 3 μL of 2× Gibson Assembly Master Mix (purchased from NEB, containing T5 exonuclease, Phusion DNA polymerase, Taq DNA ligase) was then added, and reacted at 50° C. for 1 h.

(72) The assembled product (6 μL) was used to transform 40 μL competent E. coli ER2566 (purchased from New England Biolabs) prepared by the Calcium chloride method. The transformed E. coli were spread onto solid LB medium (components of LB medium: 10 g/L peptone, 5 g/L yeast powder, 10 g/L NaCl, the same hereinafter) containing kanamycin (at a final concentration of 25 μg/mL, the same hereinafter), and were subjected to static culture at 37° C. for 10-12 h until single colonies could be observed clearly. Single colony was picked and inoculated into a tube containing 4 mL liquid LB medium (containing kanamycin), and cultured with shaking at 220 rpm for 10 h at 37° C., and then 1 ml bacterial solution was taken and stored at −70° C. Plasmids were extracted from E. coli, and T7 primer was used to sequence the nucleotide sequences of the fragments of interest inserted into the plasmids. The sequencing result showed that the nucleotide sequences of the fragments of interest inserted into the constructed plasmids (expression vectors) were SEQ ID NO: 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37 and 38, respectively, and their encoded amino acid sequences were SEQ ID NO: 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18 and 19, respectively (the corresponding proteins were designated as H18N65-45T1, H18N65-45T2, H18N65-45T3, H18N65-45T4, H18N65-45T5, H18N65-45T3-5951, H18N65-45T3-5952, H18N65-45T3-5954, H18N65-45T3-5955, H18N65-45T4- 5951, H18N65-45T4-5952, H18N65-45T4-5953, H18N65-45T4-5955, H18N65-45T1-5955, H18N65-45T2- 5955 and H18N65-45T1T3-5955, respectively).

(73) The mutated protein H18N65-45T1 differs from HPV18N65 by: the substitution of the amino acid residues from positions 114-123 of wild type HPV18 L1 protein with the amino acid residues from positions 79-89 of wild type HPV45 L1 protein. The mutated protein H18N65-45T2 differs from HPV18N65 by: the substitution of the amino acid residues from positions 176-202 of wild type HPV18 L1 protein with the amino acid residues from positions 142-168 of wild type HPV45 L1 protein. The mutated protein H18N65-45T3 differs from HPV18N65 by: the substitution of the amino acid residues from positions 235-243 of wild type HPV18 L1 protein with the amino acid residues from positions 201-209 of wild type HPV45 L1 protein. The mutated protein H18N65-45T4 differs from HPV18N65 by: the substitution of the amino acid residues from positions 327-346 of wild type HPV18 L1 protein with the amino acid residues from positions 293-314 of wild type HPV45 L1 protein. The mutated protein H18N65-45T5 differs from HPV18N65 by: the substitution of the amino acid residues from positions 411-419 of wild type HPV18 L1 protein with the amino acid residues from positions 379-387 of wild type HPV45 L1 protein.

(74) The mutated protein H18N65-45T3-5951 differs from HPV18N65 by: the substitution of the amino acid residues from positions 235-243 of wild type HPV18 L1 protein with the amino acid residues from positions 201-209 of wild type HPV45 L1 protein, and the substitution of the amino acid residues from positions 112-123 of wild type HPV18 L1 protein with the amino acid residues from positions 51-62 of wild type HPV59 L1 protein. The mutated protein H18N65-45T3-59S2 differs from HPV18N65 by: the substitution of the amino acid residues from positions 235-243 of wild type HPV18 L1 protein with the amino acid residues from positions 201-209 of wild type HPV45 L1 protein, and the substitution of the amino acid residues from positions 183-204 of wild type HPV18 L1 protein with the amino acid residues from positions 122-143 of wild type HPV59 L1 protein. The mutated protein H18N65-45T3-59S4 differs from HPV18N65 by: the substitution of the amino acid residues from positions 235-243 of wild type HPV18 L1 protein with the amino acid residues from positions 201-209 of wild type HPV45 L1 protein, and the substitution of the amino acid residues from positions 325-351 of wild type HPV18 L1 protein with the amino acid residues from positions 264-290 of wild type HPV59 L1 protein. The mutated protein H18N65-45T3-59S5 differs from HPV18N65 by: the substitution of the amino acid residues from positions 235-243 of wild type HPV18 L1 protein with the amino acid residues from positions 201-209 of wild type HPV45 L1 protein, and the substitution of the amino acid residues from positions 410-421 of wild type HPV18 L1 protein with the amino acid residues from positions 349-360 of wild type HPV59 L1 protein.

(75) The mutated protein H18N65-45T4-59S1 differs from HPV18N65 by: the substitution of the amino acid residues from positions 327-346 of wild type HPV18 L1 protein with the amino acid residues from positions 293-314 of wild type HPV45 L1 protein, and the substitution of the amino acid residues from positions 112-123 of wild type HPV18 L1 protein with the amino acid residues from positions 51-62 of wild type HPV59 L1 protein. The mutated protein H18N65-45T4-59S2 differs from HPV18N65 by: the substitution of the amino acid residues from positions 327-346 of wild type HPV18 L1 protein with the amino acid residues from positions 293-314 of wild type HPV45 L1 protein, and the substitution of the amino acid residues from positions 183-204 of wild type HPV18 L1 protein with the amino acid residues from positions 122-143 of wild type HPV59 L1 protein. The mutated protein H18N65-45T4-59S3 differs from HPV18N65 by: the substitution of the amino acid residues from positions 327-346 of wild type HPV18 L1 protein with the amino acid residues from positions 293-314 of wild type HPV45 L1 protein, and the substitution of the amino acid residues from positions 231-242 of wild type HPV18 L1 protein with the amino acid residues from positions 170-181 of wild type HPV59 L1 protein. The mutated protein H18N65-45T4-59S5 differs from HPV18N65 by: the substitution of the amino acid residues from positions 327-346 of wild type HPV18 L1 protein with the amino acid residues from positions 293-314 of wild type HPV45 L1 protein, and the substitution of the amino acid residues from positions 410-421 of wild type HPV18 L1 protein with the amino acid residues from positions 349-360 of wild type HPV59 L1 protein.

(76) The mutated protein H18N65-45T1-59S5 differs from HPV18N65 by: the substitution of the amino acid residues from positions 114-123 of wild type HPV18 L1 protein with the amino acid residues from positions 79-89 of wild type HPV45 L1 protein, and the substitution of the amino acid residues from positions 410-421 of wild type HPV18 L1 protein with the amino acid residues from positions 349-360 of wild type HPV59 L1 protein. The mutated protein H18N65-45T2-59S5 differs from HPV18N65 by: the substitution of the amino acid residues from positions 176-202 of wild type HPV18 L1 protein with the amino acid residues from positions 142-168 of wild type HPV45 L1 protein, and the substitution of the amino acid residues from positions 410-421 of wild type HPV18 L1 protein with the amino acid residues from positions 349-360 of wild type HPV59 L1 protein. The mutated protein H18N65-45T1T3-59S5 differs from HPV18N65 by: the substitution of the amino acid residues from positions 114-123 of wild type HPV18 L1 protein with the amino acid residues from positions 79-89 of wild type HPV45 L1 protein, and the substitution of the amino acid residues from positions 235-243 of wild type HPV18 L1 protein with the amino acid residues from positions 201-209 of wild type HPV45 L1 protein, and the substitution of the amino acid residues from positions 410-421 of wild type HPV18 L1 protein with the amino acid residues from positions 349-360 of wild type HPV59 L1 protein.

(77) TABLE-US-00054 TABLE 2 PCR templates and primers for constructing expression vectors Temperature/ Time of Template Upstream primer Downstream primer Product annealing 18L1N65 G-V-H18N65- G-V-H18N65- H18N65-45T1 56° C./50 s 45T1-F 45T1-R long fragment 45L1N27 G-H18N65- G-H18N65- H18N65-45T1 56° C./30 s 45T1-F 45T1-R short fragment 18L1N65 G-V-H18N65- G-V-H18N65- H18N65-45T2 56° C./50 s 45T2-F 45T2-R long fragment 45L1N27 G-H18N65- G-H18N65- H18N65-45T2 56° C./30 s 45T2-F 45T2-R short fragment 18L1N65 G-V-H18N65- G-V-H18N65- H18N65-45T3 56° C./50 s 45T3-F 45T3-R long fragment 45L1N27 G-H18N65- G-H18N65- H18N65-45T3 56° C./30 s 45T3-F 45T3-R short fragment 18L1N65 G-V-H18N65- G-V-H18N65- H18N65-45T4 56° C./50 s 45T4-F 45T4-R long fragment 45L1N27 G-H18N65- G-H18N65- H18N65-45T4 56° C./30 s 45T4-F 45T4-R short fragment 18L1N65 G-V-H18N65- G-V-H18N65- H18N65-45T5 56° C./50 s 45T5-F 45T5-R long fragment 45L1N27 G-H18N65- G-H18N65- H18N65-45T5 56° C./30 s 45T5-F 45T5-R short fragment H18N65-45T3 G-V-H18N65- G-V-H18N65- H18N65-45T3-59S1 56° C./50 s 45T3-59S1-F 45T3-59S1-R long fragment 59L1 G-H18N65- G-H18N65- H18N65-45T3-59S1 56° C./30 s 45T3-59S1-F 45T3-59S1-R short fragment H18N65-45T3 G-V-H18N65- G-V-H18N65- H18N65-45T3-59S2 56° C./50 s 45T3-59S2-F 45T3-59S2-R long fragment 59L1 G-H18N65- G-H18N65- H18N65-45T3-59S2 56° C./30 s 45T3-59S2-F 45T3-59S2-R short fragment H18N65-45T3 G-V-H18N65- G-V-H18N65- H18N65-45T3-59S4 56° C./50 s 45T3-59S4-F 45T3-59S4-R long fragment 59L1 G-H18N65- G-H18N65- H18N65-45T3-59S4 56° C./30 s 45T3-59S4-F 45T3-59S4-R short fragment H18N65-45T3 G-V-H18N65- G-V-H18N65- H18N65-45T3-59S5 56° C./50 s 45T3-59S5-F 45T3-59S5-R long fragment 59L1 G-H18N65- G-H18N65- H18N65-45T3-59S5 56° C./30 s 45T3-59S5-F 45T3-59S5-R short fragment H18N65-45T4 G-V-H18N65- G-V-H18N65- H18N65-45T4-59S1 56° C./50 s 45T4-59S1-F 45T4-59S1-R long fragment 59L1 G-H18N65- G-H18N65- H18N65-45T4-59S1 56° C./30 s 45T4-59S1-F 45T4-59S1-R short fragment H18N65-45T4 G-V-H18N65- G-V-H18N65- H18N65-45T4-59S2 56° C./50 s 45T4-59S2-F 45T4-59S2-R long fragment 59L1 G-H18N65- G-H18N65- H18N65-45T4-59S2 56° C./30 s 45T4-59S2-F 45T4-59S2-R short fragment H18N65-45T4 G-V-H18N65- G-V-H18N65- H18N65-45T4-59S3 56° C./50 s 45T4-59S3-F 45T4-59S3-R long fragment 59L1 G-H18N65- G-H18N65- H18N65-45T4-59S3 56° C./30 s 45T4-59S3-F 45T4-59S3-R short fragment H18N65-45T4 G-V-H18N65- G-V-H18N65- H18N65-45T4-59S5 56° C./50 s 45T4-59S5-F 45T4-59S5-R long fragment 59L1 G-H18N65- G-H18N65- H18N65-45T4-59S5 56° C./30 s 45T4-59S5-F 45T4-59S5-R short fragment H18N65-45T1 G-V-H18N65-4 G-V-H18N65- H18N65-45T1-59S5 56° C./50 s 5T1-59S5-F 45T1-59S5-R long fragment 59L1 G-H18N65- G-H18N65- H18N65-45T1-59S5 56° C./30 s 45T1-59S5-F 45T1-59S5-R short fragment H18N65-45T2 G-V-H18N65- G-V-H18N65- H18N65-45T2-59S5 56° C./50 s 45T2-59S5-F 45T2-59S5-R long fragment 59L1 G-H18N65- G-H18N65- H18N65-45T2-59S5 56° C./30 s 45T2-59S5-F 45T2-59S5-R short fragment H18N65-45T3- G-V-H18N65- G-V-H18N65- H18N65-45T1T3-59S5 56° C./50 s 59S5 45T1T3-59S5-F 45T1T3-59S5-R long fragment 45L1N27 G-H18N65- G-H18N65- H18N65-45T1T3-59S5 56° C./30 s 45T1T3-59S5-F 45T1T3-59S5-R short fragment

(78) TABLE-US-00055 TABLE 3 Sequences of the primers used (SEQ ID NOs: 45-108) SEQ ID NO: Primer name Primer sequence (5′-3′) 45 G-V-H18N65-45T1-F CAGTTACCTGACCCAAATAAATT 46 G-V-H18N65-45T1-R AGTCACGTAATCATCGGTAT 47 G-H18N65-45T1-F TAAATACCGATGATTACGTGACTAGGACCAGCATCTT CTACCAC 48 G-H18N65-45T1-F AATTTATTTGGGTCAGGTAACTGCACCCTGAACACCC TGTACTGG 49 G-V-H18N65-45T2-F GCCCCTGCTATTGGGGAACACTGGGCT 50 G-V-H18N65-45T2-R CACTCCAGCACAGGCCCACACTAAAC 51 G-H18N65-45T2-F GTGTGGGCCTGTGCTGGAGTGGAGATCGGCAGGGGC CAG 52 G-H18N65-45T2-R CAGTGTTCCCCAATAGCAGGGGCGCAGCCCAGGATG CACAGCT 53 G-V-H18N65-45T3-F GTTTTGGAAGATGGTGATATGGT 54 G-V-H18N65-45T3-R GGCACAGCCCAAAATACATAACT 55 G-H18N65-45T3-F AGTTATGTATTTTGGGCTGTGCCCCCGCCATCGGCGA GCACTGGG 56 G-H18N65-45T3-R ACCATATCACCATCTTCCAAAACGGTGTTCTTCAGCT CCAGGGG 57 G-V-H18N65-45T4-F GTTACCTCTGACTCCCAGTTGTT 58 G-V-H18N65-45T4-R TCCAGTATCTACCATATCACCATCTT 59 G-H18N65-45T4-F GATGGTGATATGGTAGATACTGGATACGGCGCCATG GACTTCAGCAC 60 G-H18N65-45T4-R AACAACTGGGAGTCAGAGGTAACGATGCTGCCGCTG GGGCTGGGGCT 61 G-V-H18N65-45T5-F GATGTTATGTCCTATATTCAT 62 G-V-H18N65-45T5-R TATTGTTAAATTGGTACTGCGAG 63 G-H18N65-45T5-F CTCGCAGTACCAATTTAACAATATGCGCCAGCACCC AGAACCCCG 64 G-H18N65-45T5-R CTATGAATATAGGACATAACATCGGCGGTCAGGGTG ATGGTGCAC 65 G-V-H18N65-45T3-59S1-F GCATACCAATATAGAGTATTTAG 66 G-V-H18N65-45T3-59S1-R ATATGGATTACCAACAGTTAATAAT 67 G-H18N65-45T3-59S1-F TATTAACTGTTGGTAATCCATATTTCAAGGTGCCCAA GGGCGGC 68 G-H18N65-45T3-59S1-R CCTAAATACTCTATATTGGTATGCGCTCACCTTGGGC ACGTCCTGC 69 G-V-H18N65-45T3-59S2-F TTGGGCTGTGCCCCCGCCATCGG 70 G-V-H18N65-45T3-59S2-R AGCACAGGCCCACACTAAACGTTGT 71 G-H18N65-45T3-59S2-F CAACGTTTAGTGTGGGCCTGTGCTGGCGTGGAGATC GGCAGGGGC 72 G-H18N65-45T3-59S2-R GCCGATGGCGGGGGCACAGCCCAAGATGCACAGCTG GGTCTGCTTGT 73 G-V-H18N65-45T3-59S4-F GGCTCTATTGTTACCTCTGACTC 74 G-V-H18N65-45T3-59S4-R AAGCTGCTCACGCCGTAAGCAAAAA 75 G-H18N65-45T3-59S4-F TGCTTACGGCGTGAGCAGCTTTTCGCCAGGCACTTCT GGAACAG 76 G-H18N65-45T3-59S4-R GGAGTCAGAGGTAACAATAGAGCCGCTGGGGCTGGG GCTGTACAGGT 77 G-V-H18N65-45T3-59S5-F CAGTATAGCAGACATGTTGAGG 78 G-V-H18N65-45T3-59S5-R TATTGTTAAATTGGTACTGCGAG 79 G-H18N65-45T3-59S5-F ACTCGCAGTACCAATTTAACAATATGCGCCAGCACC ACCAGCAGCAT 80 G-H18N65-45T3-59S5-R TTCCTCAACATGTCTGCTATACTGCTTGAAGCTGGTG GGGGTGT 81 G-V-H18N65-45T4-59S1-F GCATACCAATATAGAGTATTTAG 82 G-V-H18N65-45T4-59S1-R ATTACCAACAGTTAATAATCTAGAGC 83 G-H18N65-45T4-59S1-F GATTATTAACTGTTGGTAATCCCTACTTCAAGGTGCC CAAGGGCGG 84 G-H18N65-45T4-59S1-R ATTTGGGTCAGGTAACTGCACCCTGAACACCCTGTAC TGGTAGGCGC 85 G-V-H18N65-45T4-59S2-F TTATGTATTTTGGGCTGTGCCCCTG 86 G-V-H18N65-45T4-59S2-R AGCACAGGCCCACACTAAACGTT 87 G-H18N65-45T4-59S2-F GTTTAGTGTGGGCCTGTGCTGGCGTGGAGATCGGCA GGGGCCAGCCC 88 G-H18N65-45T4-59S2-R GCAGGGGCACAGCCCAAAATACATAACTGGGTCTGC TTGTAGTCCAC 89 G-V-H18N65-45T4-59S3-F AAAAACACAGTTTTGGAAGATGGTG 90 G-V-H18N65-45T4-59S3-R GGCACAGCCCAAAATACATAACT 91 G-H18N65-45T4-59S3-F TTATGTATTTTGGGCTGTGCCCCCGCCATCGGCGAGC ACTGGAC 92 G-H18N65-45T4-59S3-R ACCATCTTCCAAAACTGTGTTTTTCAGCTCCAGGGGG GGGCAGTCGC 93 G-V-H18N65-45T4-59S5-F CAGTATAGCAGACATGTTGAGG 94 G-V-H18N65-45T4-59S5-R TATTGTTAAATTGGTACTGCGGTGGT 95 G-H18N65-45T4-59S5-F CAGTACCAATTTAACAATATGCGCCAGCACCACCAG CAGCATCCCC 96 G-H18N65-45T4-59S5-R ATATTCCTCAACATGTCTGCTATACTGCTTGAAGCTG GTGGGGGTGT 97 G-V-H18N65-45T1-59S5-F CAGTATAGCAGACATGTTGAGG 98 G-V-H18N65-45T1-59S5-R TATTGTTAAATTGGTACTGCGAG 99 G-H18N65-45T1-59S5-F ACTCGCAGTACCAATTTAACAATATGCGCCAGCACC ACCAGCAGCAT 100 G-H18N65-45T1-59S5-R TTCCTCAACATGTCTGCTATACTGCTTGAAGCTGGTG GGGGTGT 101 G-V-H18N65-45T2-59S5-F CAGTATAGCAGACATGTTGAGG 102 G-V-H18N65-45T2-59S5-R TATTGTTAAATTGGTACTGCGAG 103 G-H18N65-45T2-59S5-F ACTCGCAGTACCAATTTAACAATATGCGCCAGCACC ACCAGCAGCAT 104 G-H18N65-45T2-59S5-R TTCCTCAACATGTCTGCTATACTGCTTGAAGCTGGTG GGGGTGT 105 G-V-H18N65-45T1T3-59S5-F CAGTTACCTGACCCAAATAAATT 106 G-V-H18N65-45T1T3-59S5-R AGTCACGTAATCATCGGTAT 107 G-H18N65-45T1T3-59S5-F TAAATACCGATGATTACGTGACTAGGACCAGCATCTT CTACCAC 108 G-H18N65-45T1T3-59S5-R AATTTATTTGGGTCAGGTAACTGCACCCTGAACACCC TGTACTGG

(79) Expression of the Mutated Proteins on a Large Scale

(80) The E. coli solutions comprising the recombinant plasmid pTO-T7-H18N65-45T1, pTO-T7-H18N65-45T2, pTO-T7-H18N65-45T3, pTO-T7-H18N65-45T4, pTO-T7-H18N65-45T5, pTO-T7-H18N65-45T3-59S1, pTO-T7-H18N65-45T3-59S2, pTO-T7-H18N65-45T3-59S4, pTO-T7-H18N65-45T3-59S5, pTO-T7-H18N65-45T4-59S1, pTO-T7-H18N65-45T4-59S2, pTO-T7-H18N65-45T4-59S3, pTO-T7-H18N65-45T4-59S5, pTO-T7-H18N65-45T1-59S5, pTO-T7-H18N65-45T2-59S5 and pTO-T7-H18N65-45T1T3-59S5, respectively, were taken from −70° C. refrigerator, were inoculated in 100 mL LB liquid medium containing kanamycin, and incubated at 200 rpm and 37° C. for about 8 h. Then, the culture was transferred to 500 mL LB medium containing kanamycin (1 ml bacterial solution was transferred), and was further incubated. When the bacterial concentration reached an OD.sub.600 of about 0.6, the culturing temperature was lowered to 25° C. and 500 μL IPTG was added to each culture bottle. The incubation was further performed for 8 h. After the incubation was finished, the bacteria were collected by centrifugation. The bacteria expressing H18N65-45T1, H18N65-45T2, H18N65-45T3, H18N65-45T4, H18N65-45T5, H18N65-45T3-59S1, H18N65-45T3-59S2, H18N65-45T3-59S4, H18N65-45T3-59S5, H18N65-45T4-59S1, H18N65-45T4-59S2, H18N65-45T4-59S3, H18N65-45T4-59S5, H18N65-45T1-59S5, H18N65-45T2-59S5 and H18N65-45T1T3-59S5 protein were obtained, respectively.

(81) Disruption of Bacteria Expressing the Mutated Proteins

(82) The bacteria obtained above were re-suspended at a ratio of 1 g bacteria to 10 mL lysis buffer (20 mM Tris buffer, pH7.2, 300 mM NaCl). The bacteria were disrupted by using an ultrasonic apparatus for 30 min. The lysis solution containing the disrupted bacteria were centrifuged at 13500 rpm (30000 g) for 15 min, and the supernatant (i.e. the supernatant of disrupted bacteria) was obtained.

(83) Chromatographic Purification of the Mutated Protein

(84) Equipment: AKTA Explorer 100 preparative liquid chromatography system produced by GE Healthcare (i.e. the original Amershan Pharmacia Co.)

(85) Chromatographic media: SP Sepharose 4 Fast Flow (GE Healthcare Co.), CHT-II (purchased from Bio-RAD) and Butyl Sepharose 4 Fast Flow (GE Healthcare Co.)

(86) Buffer: Buffer A (20 mM phosphate buffer, pH8.0, 20 mM DTT); and Buffer B (20 mM phosphate buffer, pH8.0, 20 mM DTT, 2M NaCl). The buffers containing different concentrations of NaCl used in the following elution protocol were prepared by mixing Buffer A and Buffer B at a certain ratio.

(87) Sample: the supernatants of disrupted bacteria containing H18N65-45T1, H18N65-45T2, H18N65-45T3, H18N65-45T4, H18N65-45T5, H18N65-45T3-5951, H18N65-45T3-5952, H18N65-45T3-59S4, H18N65-45T3-5955, H18N65-45T4-5951, H18N65-45T4-5952, H18N65-45T4-5953, H18N65-45T4-59S5, H18N65-45T1-59S5, H18N65-45T2-5955, and H18N65-45T1T3-5955, respectively, as obtained above.

(88) Elution Protocol:

(89) (1) Cation exchange purification of the supernatant of disrupted bacteria by SP Sepharose 4 Fast Flow: the sample was loaded on the column, undesired proteins were then eluted with a buffer containing 400 mM NaCl (80% Buffer A+20% Buffer B), followed by the elution of the protein of interest with a buffer containing 800 mM NaCl (60% Buffer A+40% Buffer B), and the fraction eluted with the buffer containing 800 mM NaCl was collected;

(90) (2) Chromatographic purification of the elution fraction obtained in the step (1) by CHTII (hydroxyapatite chromatography): the elution fraction obtained in the step (1) was diluted so that the NaCl concentration was decreased to 0.5 M; the sample was loaded on the column, undesired proteins were then eluted with a buffer containing 500 mM NaCl (75% Buffer A+25% Buffer B), followed by the elution of the protein of interest with a buffer containing 1000 mM NaCl (50% Buffer A+50% Buffer B), and the fraction eluted with the buffer containing 1000 mM NaCl was collected;

(91) (3) Chromatographic purification of the elution fraction obtained in the step (2) by HIC (hydrophobic interaction chromatography): the sample was loaded on the column, undesired proteins were then eluted with a buffer containing 1000 mM NaCl, followed by the elution of the protein of interest with a buffer containing 200 mM NaCl (90% Buffer A+10% Buffer B), and the fraction eluted with the buffer containing 200 mM NaCl was collected.

(92) 150 μL of elution fraction obtained in the step (3) was added to 30 μL of 6× Loading Buffer (1 L of which contained 300 ml of 1M TB 6.8, 600 ml of 100% glycerol, 120 g of SDS, 6 g of bromophenol blue, and 50 ml of β-mercaptoethanol). The resultant solution was mixed well and incubated in 80° C. water bath for 10 min. 10 μl of the resultant sample was then subjected to 10% SDS-PAGE at 120V for 120 min; and the electrophoretic bands were stained by Coomassie brilliant blue. The electrophoretic result was shown in FIG. 1. The result showed that after said purification steps, H18N65-45T1, H18N65-45T2, H18N65-45T3, H18N65-45T4, H18N65-45T5, H18N65-45T3-59S1, H18N65-45T3-59S2, H18N65-45T3-59S4, H18N65-45T3-59S5, H18N65-45T4-59S1, H18N65-45T4-59S2, H18N65-45T4-59S3, H18N65-45T4-59S5, H18N65-45T1-59S5, H18N65-45T2-59S5 and H18N65-45T1T3-59S5 protein had a purity of above 85%.

(93) By similar methods, HPV18N65 protein (SEQ ID NO: 113) was prepared and purified by using E. coli and the plasmid pTO-T7-HPV18N65L1; HPV45N27 protein (SEQ ID NO: 115) was prepared and purified by using E. coli and the plasmid pTO-T7-HPV45L1N27C; and HPV59 L1 protein (SEQ ID NO: 3) was prepared and purified by using E. coli and the plasmid pTO-T7-HPV59L1.

(94) Western Blot Assay of the Mutated Proteins

(95) The H18N65-45T1, H18N65-45T2, H18N65-45T3, H18N65-45T4, H18N65-45T5, H18N65-45T3-5951, H18N65-45T3-5952, H18N65-45T3-5954, H18N65-45T3-5955, H18N65-45T4-5951, H18N65-45T4-5952, H18N65-45T4-5953, H18N65-45T4-5955, H18N65-45T1-5955, H18N65-45T2-5955, and H18N65-45T1T3-59S5 protein purified by the method above were subjected to electrophoresis. After electrophoresis, Western Blot assay was carried out by using a broad-spectrum antibody 4B3 against HPV L1 protein, and the result was shown in FIG. 2. The result showed that H18N65-45T1, H18N65-45T2, H18N65-45T3, H18N65-45T4, H18N65-45T5, H18N65-45T3-5951, H18N65-45T3-5952, H18N65-45T3-5954, H18N65-45T3-5955, H18N65-45T4-5951, H18N65-45T4-5952, H18N65-45T4-5953, H18N65-45T4- 5955, H18N65-45T1-5955, H18N65-45T2-5955 and H18N65-45T1T3-5955 could be specifically recognized by the broad-spectrum antibody 4B3.

Example 2: Assembly of HPV Virus-Like Particles and Morphological Detection of Particles

(96) Assembly of HPV Virus-Like Particles

(97) A given volume (about 2 ml) of the protein H18N65-45T1, H18N65-45T2, H18N65-45T3, H18N65-45T4, H18N65-45T5, H18N65-45T3-5951, H18N65-45T3-5952, H18N65-45T3-5954, H18N65-45T3-5955, H18N65-45T4-5951, H18N65-45T4-5952, H18N65-45T4-5953, H18N65-45T4-5955, H18N65-45T1-5955, H18N65-45T2-5955, or H18N65-45T1T3-5955, was dialyzed to (1) 2 L storage buffer (20 mM sodium phosphate buffer pH 6.5, 0.5 M NaCl); (2) 2 L renaturation buffer (50 mM sodium phosphate buffer pH 6.0, 2 mM CaCl.sub.2), 2 mM MgCl.sub.2, 0.5 M NaCl); and (3) 20 mM sodium phosphate buffer pH 7.0, 0.5 M NaCl, successively. The dialysis was performed in each of the three buffers for 12 h.

(98) By similar methods, the HPV18N65, HPV45N27 and HPV59 L1 protein were assembled into HPV18N65 VLP, HPV45N27 VLP and HPV59 VLP, respectively.

(99) Molecular Sieve Chromatographic Analysis

(100) The dialyzed sample was subjected to molecular sieve chromatographic analysis by 1120 Compact LC High Performance Liquid Chromatographic System (Agilent Technologies), wherein the analytical column used was TSK Gel PW5000×1 7.8×300 mm. The analysis results were shown in FIGS. 3A-3S. The results showed that the first protein peak of the samples comprising the protein H18N65-45T1, H18N65-45T2, H18N65-45T3, H18N65-45T4, H18N65-45T5, H18N65-45T3-5951, H18N65-45T3-5952, H18N65-45T3-5954, H18N65-45T3-5955, H18N65-45T4-5951, H18N65-45T4-5952, H18N65-45T4- 5953, H18N65-45T4-5955, H18N65-45T1-5955, H18N65-45T2-5955 or H18N65-45T1T3-5955 appeared at about 13 min, which was comparable to that of HPV18N65 VLP, HPV45N27 VLP and HPV59 VLP. This showed that all these protein were able to assemble into VLPs.

(101) Morphological Test of Virus-Like Particles

(102) A 100 μL sample comprising VLP was observed by transmission electron microscope (TEM). The apparatus used was a 100 kV Transmission Electron Microscope supplied by JEOL Ltd. (100,000× magnification). In brief, a 13.5 μL of sample was negatively stained with 2% phosphotungstic acid (pH 7.0), fixed on a carbon-coated copper grid, and then observed by TEM. The results were shown in FIGS. 4A-4S. The results showed that H18N65-45T1, H18N65-45T2, H18N65-45T3, H18N65-45T4, H18N65-45T5, H18N65-45T3-5951, H18N65-45T3-5952, H18N65-45T3-5954, H18N65-45T3-5955, H18N65-45T4- 5951, H18N65-45T4-5952, H18N65-45T4-5953, H18N65-45T4-5955, H18N65-45T1-5955, H18N65-45T2- 5955 and H18N65-45T1T3-5955 were able to assemble into virus-like particles. In addition, the results also showed that the particles assembled by these mutated proteins had a radius of about 30 nm, and were uniform in size. The particles assembled by wild type HPV18N65, HPV45N27 and HPV59 L1 also had a radius of about 30 nm, and were uniform in size. This indicated that these mutated proteins were similar to the L1 protein of HPV18, HPV45 and HPV59, and were able to assemble into VLPs with a uniform size.

(103) Sedimentation Velocity Analysis

(104) The apparatus for sedimentation velocity analysis was Beckman XL-A Analytical Ultracentrifuge, equipped with optical inspection system and An-50Ti and An-60Ti rotor. The sedimentation coefficients of HPV18N65 VLP, HPV45N27 VLP, HPV59 VLP, H18N65-45T3 VLP, H18N65-45T4 VLP, H18N65-45T3-59S1 VLP, H18N65-45T4-59S1 VLP and H18N65-45T1T3-59S5 VLP were analyzed by sedimentation velocity method. The results were shown in FIGS. 5A-5H. The results showed that the sedimentation coefficient of H18N65-45T3 VLP, H18N65-45T4 VLP and H18N65-45T1T3-59S5 VLP was 143.7S, 173.3S and 167.1S, respectively, which was similar to that of HPV18N65 VLP, HPV45N27 VLP and HPV59 VLP (HPV18N65 VLP, 142.2S; HPV45N27 VLP, 146.5S, and HPV59 VLP, 139.3S). This showed that H18N65-45T3 VLP, H18N65-45T4 VLP and H18N65-45T1T3-59S5 VLP were able to assemble into virus-like particles that were similar to wild type VLP in terms of size and morphology.

Example 3: Evaluation of Thermostability of Virus-Like Particles

(105) The VLPs formed by protein HPV18N65, HPV45N27, HPV59, H18N65-45T3, H18N65-45T4, H18N65-45T3-59S1, H18N65-45T4-59S1, and H18N65-45T1T3-59S5 were evaluated for their thermostability by using a differential scanning calorimeter VP Capillary DSC purchased from GE Company (i.e. the original MicroCal Co.), wherein the storage buffer for the protein was used as control, and the proteins were scanned at a heating rate of 1.5° C./min within a temperature range of 10° C.-90° C. The detection results were shown in FIGS. 6A-6H. The results showed that all these VLPs formed by the proteins had very high thermostability.

Example 4: Evaluation 1 of Neutralizing Antibody Titer in Serum of Mice Vaccinated with Virus-Like Particles

(106) The immune protection of the VLPs formed by H18N65-45T1, H18N65-45T2, H18N65-45T3, H18N65-45T4, H18N65-45T5, H18N65-45T3-59S1, H18N65-45T3-59S2, H18N65-45T3-59S4, H18N65-45T3-59S5, H18N65-45T4-59S1, H18N65-45T4-59S2, H18N65-45T4-59S3, H18N65-45T4-59S5, H18N65-45T1-59S5, H18N65-45T2-59S5 and H18N65-45T1T3-59S5 was evaluated in mice. Animals for vaccination were BALB/c mice (ordinary grade), 5-6 weeks old (purchased from Shanghai SLAC Laboratory Animal Co. LTD.).

(107) The H18N65-45T1 VLP, H18N65-45T2 VLP, H18N65-45T3 VLP, H18N65-45T4 VLP, H18N65-45T5 VLP, HPV18N65 VLP, HPV45N27 VLP and a mixed HPV18/HPV45 VLP (i.e. a mixture of HPV18N65 VLP and HPV45N27 VLP) as prepared above were absorbed onto aluminum adjuvant, respectively. Mice were divided into 8 groups depending on immunogen, and each group included 5 mice. Vaccination procedure was as followed: the first vaccination at Week 0, and the booster vaccination at Weeks 2 and 4, respectively. Mice were vaccinated via intraperitoneal injection. The immunogens and doses thereof were shown in Table 4. At Week 8 after the first vaccination, venous blood was collected from eyeball, and serum was separated. The titers of neutralizing antibodies in the serum were determined. The detection result was shown in FIG. 7A. The result showed that H18N65-45T1 VLP, H18N65-45T2, VLPH18N65-45T3 VLP and H18N65-45T4 VLP each retained the activity that could induce the generation of high-titer neutralizing antibodies against HPV18 in mice, and their protective effects against HPV18 were comparable to that of HPV18N65 VLP alone and that of the mixed HPV18/HPV45 VLP, and their activities that could induce the generation of neutralizing antibodies against HPV45 in mice were higher than that of HPV18N65 VLP alone. In particular, H18N65-45T3 VLP and H18N65-45T4 VLP each could induce the generation of high-titer neutralizing antibodies against HPV45 and HPV18 in mice; and their protective effects against HPV18 were comparable to that of HPV18N65 VLP alone and that of the mixed HPV18/HPV45 VLP, and were significantly higher than that of HPV45N27 VLP alone; and their protective effects against HPV45 were comparable to that of HPV45N27 VLP alone and that of the mixed HPV18/HPV45 VLP, and were significantly higher than that of HPV18N65 VLP alone. This showed that after mutation, H18N65-45T1 VLP, H18N65-45T2, VLPH18N65-45T3 VLP and H18N65-45T4 VLP retained their immunogenicity against HPV18, and their immunogenicity against HPV45 is also improved as compared to HPV18N65 VLP. Especially, H18N65-45T3 VLP and H18N65-45T4 VLP had good cross-immunogenicity and cross-protection against HPV18 and HPV45, and could be used as effective vaccines for preventing HPV18 infection and/or HPV45 infection, and could be used in place of a mixed vaccine comprising HPV18 VLP and HPV45 VLP.

(108) TABLE-US-00056 TABLE 4 Vaccination schedule Vaccination Antigen for Immunizing procedure vaccination Adjuvant dose Number (week) H18N65-45T1 VLP aluminum 5 μg 5 0, 2, 4 adjuvant H18N65-45T2 VLP aluminum 5 μg 5 0, 2, 4 adjuvant H18N65-45T3 VLP aluminum 5 μg 5 0, 2, 4 adjuvant H18N65-45T4 VLP aluminum 5 μg 5 0, 2, 4 adjuvant H18N65-45T5 VLP aluminum 5 μg 5 0, 2, 4 adjuvant HPV18N65 VLP aluminum 5 μg 5 0, 2, 4 adjuvant HPV45N27 VLP aluminum 5 μg 5 0, 2, 4 adjuvant HPV18/HPV45 VLP aluminum 5 μg for 5 0, 2, 4 adjuvant each VLP

(109) In addition, the H18N65-45T3-59S1 VLP, H18N65-45T3-59S2 VLP, H18N65-45T3-59S4 VLP, H18N65-45T3-59S5 VLP, H18N65-45T4-59S1 VLP, H18N65-45T4-59S2 VLP, H18N65-45T4-59S3 VLP, H18N65-45T4-59S5 VLP, HPV18N65 VLP, HPV45N27 VLP, HPV59 VLP and the mixed HPV18/HPV45/HPV59 VLP (i.e. a mixture of HPV18N65 VLP, HPV45N27 VLP and HPV59 VLP) as prepared above were absorbed onto aluminum adjuvant, respectively. Mice were divided into 12 groups depending on immunogen, and each group included 5 mice. Vaccination procedure was as followed: the first vaccination at Week 0, and the booster vaccination at Weeks 2 and 4, respectively. Mice were vaccinated via intraperitoneal injection. The immunogens and doses thereof were shown in Table 5. At Week 8 after the first vaccination, venous blood was collected from eyeball, and serum was separated. The titers of neutralizing antibodies in the serum were determined. The detection result was shown in FIG. 7B. The result showed that H18N65-45T3-59S1 VLP and H18N65-45T4-59S1 VLP each could induce the generation of high-titer neutralizing antibodies against HPV18, HPV45 and HPV59 in mice; and their protective effects against HPV18 were comparable to that of HPV18N65 VLP alone and that of the mixed HPV18/HPV45/HPV59 VLP, and were significantly higher than that of HPV45N27 VLP alone and that of HPV59 VLP alone; and their protective effects against HPV45 were comparable to that of HPV45N27 VLP alone and that of the mixed HPV18/HPV45/HPV59 VLP, and were significantly higher than that of HPV18N65 VLP alone and that of HPV59 VLP alone; and their protective effects against HPV59 were comparable to that of HPV59 VLP alone and that of the mixed HPV18/HPV45/HPV59 VLP, and were significantly higher than that of HPV45N27 VLP alone and that of HPV18N65 VLP alone. This showed that H18N65-45T3-59S1 VLP and H18N65-45T4-59S1 VLP had good cross-immunogenicity and cross-protection against HPV18, HPV45 and HPV59, and could be used as effective vaccines for preventing HPV18 infection, HPV45 infection and/or HPV59 infection, and could be used in place of a mixed vaccine comprising HPV18 VLP, HPV45 VLP and HPV59 VLP.

(110) TABLE-US-00057 TABLE 5 Vaccination schedule Vaccination Immunizing procedure Antigen for vaccination Adjuvant dose Number (week) H18N65-45T3-59S1 VLP aluminum 5 μg 5 0, 2, 4 adjuvant H18N65-45T3-59S2 VLP aluminum 5 μg 5 0, 2, 4 adjuvant H18N65-45T3-59S4 VLP aluminum 5 μg 5 0, 2, 4 adjuvant H18N65-45T3-59S5 VLP aluminum 5 μg 5 0, 2, 4 adjuvant H18N65-45T4-59S1 VLP aluminum 5 μg 5 0, 2, 4 adjuvant H18N65-45T4-59S2 VLP aluminum 5 μg 5 0, 2, 4 adjuvant H18N65-45T4-59S3 VLP aluminum 5 μg 5 0, 2, 4 adjuvant H18N65-45T4-59S5 VLP aluminum 5 μg 5 0, 2, 4 adjuvant HPV18N65 VLP aluminum 5 μg 5 0, 2, 4 adjuvant HPV45N27 VLP aluminum 5 μg 5 0, 2, 4 adjuvant HPV59 VLP aluminum 5 μg 5 0, 2, 4 adjuvant HPV18/HPV45/ aluminum 5 μg for 5 0, 2, 4 HPV59 VLP adjuvant each VLP

Example 5: Evaluation of ED.SUB.50 .of Virus-Like Particles for Inducing Seroconversion

(111) In this experiment, virus-like particles used were H18N65-45T3-59S1 VLP, H18N65-45T4-59S1 VLP and H18N65-45T1T3-59S5 VLP.

(112) 6-Week old BalB/c female mice (8 mice) were vaccinated with aluminum adjuvant by single intraperitoneal injection, wherein H18N65-45T3-59S1 VLP, H18N65-45T4-59S1 VLP or H18N65-45T1T3-59S5 (at an immunizing dose of 0.300 μg, 0.100 μg, 0.033 μg, 0.011 μg or 0.004m) was used in the Experimental groups, and HPV45N27 VLP alone, HPV18N65 VLP alone, HPV59 VLP alone (at an immunizing dose of 0.300m, 0.100 μg, 0.033m, 0.011 μg or 0.004 μg) or the mixed HPV18/HPV45/HPV59 VLP (i.e. a mixture of HPV18N65 VLP, HPV45N27 VLP and HPV59 VLP, at an immunizing dose of 0.300m, 0.100 μg, 0.033m, 0.011 μg or 0.004 μg for each VLP) was used in the Control groups; the immunizing volume was 1 mL. In addition, the diluent used to dilute the vaccine was used as a blank control. 8 Mice were vaccinated in each group, and at Week 5 after vaccination, venous blood was collected from eyeball. Antibodies against HPV in the serum were detected, and by Reed-Muench method (Reed LJ MH. A simple method of estimating fifty percent endpoints. Am J Hyg. 1938; 27:493-7), ED.sub.50 for inducing seroconversion (i.e. inducing the generation of antibodies in mice) was calculated for each sample. The results were shown in Tables 6-12.

(113) TABLE-US-00058 TABLE 6 ED.sub.50 of HPV18N65 VLP for inducing the generation of antibodies against HPV45, HPV18 and HPV59 (seroconversion) in mice Number of Total mice with Positive Immunizing number positive conversion ED.sub.50 Type dose (μg) of mice conversion rate (μg) HPV18 0.300 8 6  83.33% 0.138 0.100 8 3  36.36% 0.033 8 1  6.67% 0.011 8 0  0.00% 0.004 8 0  0.00% HPV45 0.300 8 0 100.00% >0.3 0.100 8 0 100.00% 0.033 8 0  90.00% 0.011 8 0  22.22% 0.004 8 0  0.00% HPV59 0.300 8 0  0.00% >0.3 0.100 8 0  0.00% 0.033 8 0  0.00% 0.011 8 0  0.00% 0.004 8 0  0.00%

(114) TABLE-US-00059 TABLE 7 ED.sub.50 of HPV45N27 VLP for inducing the generation of antibodies against HPV45, HPV18 and HPV59 (seroconversion) in mice Number of Total mice with Positive Immunizing number positive conversion ED.sub.50 Type dose (μg) of mice conversion rate (μg) HPV18 0.300 8 0  0.00% >0.3 0.100 8 0  0.00% 0.033 8 0  0.00% 0.011 8 0  0.00% 0,004 8 0  0.00% HPV45 0.300 8 7 95.83% 0.021 0.100 8 8 94.12% 0.033 8 7 80.00% 0.011 8 1 10.00% 0.004 8 0  0.00% HPV59 0.300 8 0  0.00% >0.3 0.100 8 0  0.00% 0.033 8 0  0.00% 0.011 8 0  0.00% 0.004 8 0  0.00%

(115) TABLE-US-00060 TABLE 8 ED.sub.50 of HPV59 VLP for inducing the generation of antibodies against HPV45, HPV18 and HPV59 (seroconversion) in mice Number of Total mice with Positive Immunizing number positive conversion ED.sub.50 Type dose (μg) of mice conversion rate (μg) HPV18 0.300 8 0  0.00% >0.3 0.100 8 0  0.00% 0.033 8 0  0.00% 0.011 8 0  0.00% 0.004 8 0  0.00% HPV45 0.300 8 0  0.00% >0.3 0.100 8 0  0.00% 0.033 8 0  0.00% 0.011 8 0  0.00% 0.004 8 0  0.00% HPV59 0.300 8 8 100.00% 0.029 0.100 8 8 100.00% 0.033 8 4  55.56% 0.011 8 1  8.33% 0.004 8 0  0.00%

(116) TABLE-US-00061 TABLE 9 ED.sub.50 of the mixed HPV18/HPV45/HPV59 VLP for inducing the generation of antibodies against HPV45, HPV18 and HPV59 (seroconversion) in mice Number of Total mice with Positive Immunizing number positive conversion ED.sub.50 Type dose (μg) of mice conversion rate (μg) HPV18 0.300 μg for each VLP 8 6 86.67% 0.088 0.100 μg for each VLP 8 4 53.85% 0.033 μg for each VLP 8 3 21.43% 0.011 μg for each VLP 8 0  0.00% 0.004 μg for each VLP 8 0  0.00% HPV45 0.300 μg for each VLP 8 6 88.24% 0.033 0.100 μg for each VLP 8 6 69.23% 0.033 μg for each VLP 8 3 25.00% 0.011 μg for each VLP 8 0  0.00% 0.004 μg for each VLP 8 0  0.00% HPV59 0.300 μg for each VLP 8 6 91.30% 0.025 0.100 μg for each VLP 8 8 88.24% 0.033 μg for each VLP 8 6 63.64% 0.011 μg for each VLP 8 1  8.33% 0.004 μg for each VLP 8 0  0.00%

(117) TABLE-US-00062 TABLE 10 ED.sub.50 of H18N65-45T3-59S1 VLP for inducing the generation of antibodies against HPV45, HPV18 and HPV59 (seroconversion) in mice Number of Total mice with Positive Immunizing number positive conversion ED.sub.50 Type dose (μg) of mice conversion rate (μg) HPV18 0.300 8 7 94.12% 0.057 0.100 8 7 81.82% 0.033 8 2 20.00% 0.011 8 0  0.00% 0.004 8 0  0.00% HPV45 0.300 8 2 53.85% 0.244 0.100 8 4 33.33% 0.033 8 1  5.56% 0.011 8 0  0.00% 0.004 8 0  0.00% HPV59 0.300 8 7 90.91% 0.152 0.100 8 0 25.00% 0.033 8 2 16.67% 0.011 8 1  4.35% 0.004 8 0  0.00%

(118) TABLE-US-00063 TABLE 11 ED.sub.50 of H18N65-45T4-59S1 VLP for inducing the generation of antibodies against HPV45, HPV18 and HPV59 (seroconversion) in mice Number of Total mice with Positive Immunizing number positive conversion ED.sub.50 Type dose (μg) of mice conversion rate (μg) HPV18 0.300 8 6 75.00% 0.208 0.100 8 0  0.00% 0.033 8 0  0.00% 0.011 8 0  0.00% 0.004 8 0  0.00% HPV45 0.300 8 4 55.56% 0.264 0.100 8 0  7.69% 0.033 8 1  5.00% 0.011 8 0  0.00% 0.004 8 0  0.00% HPV59 0.300 8 6 77.78% 0.191 0.100 8 1 10.00% 0.033 8 0  0.00% 0.011 8 0  0.00% 0.004 8 0  0.00%

(119) TABLE-US-00064 TABLE 12 ED.sub.50 of H18N65-45T1T3-59S5 VLP for inducing the generation of antibodies against HPV45, HPV18 and HPV59 (seroconversion) in mice Number of Total mice with Positive Immunizing number positive conversion ED.sub.50 Type dose (μg) of mice conversion rate (μg) HPV18 0.300 8 8 100.00% 0.065 0.100 8 5  70.00% 0.033 8 2  18.18% 0.011 8 0  0.00% 0.004 8 0  0.00% HPV45 0.300 8 8 100.00% 0.086 0.100 8 3  54.55% 0.033 8 2  21.43% 0.011 8 1  5.26% 0.004 8 0  0.00% HPV59 0.300 8 8 100.00% 0.125 0.100 8 3  37.50% 0.033 8 0  0.00% 0.011 8 0  0.00% 0.004 8 0  0.00%

(120) The results showed that 5 weeks after vaccination of mice, ED.sub.50 of H18N65-45T3-59S1 VLP, H18N65-45T4-59S1 VLP and H18N65-45T1T3-59S5 VLP for inducing the generation of antibodies against HPV18 in mice was comparable to that of HPV18N65 VLP alone and that of the mixed HPV18/HPV45/HPV59 VLP, and was significantly superior to that of HPV45N27 VLP alone and that of HPV59 VLP alone; and ED.sub.50 of H18N65-45T3-59S1 VLP, H18N65-45T4-59S1 VLP and H18N65-45T1T3-59S5 VLP for inducing the generation of antibodies against HPV45 in mice was comparable to that of HPV45N27 VLP alone and that of the mixed HPV18/HPV45/HPV59 VLP, and was significantly superior to that of HPV18N65 VLP alone and that of HPV59 VLP alone; and ED.sub.50 of H18N65-45T3-59S1 VLP, H18N65-45T4-59S1 VLP and H18N65-45T1T3-59S5 VLP for inducing the generation of antibodies against HPV59 in mice was comparable to that of HPV59 VLP alone and that of the mixed HPV18/HPV45/HPV59 VLP, and was significantly superior to that of HPV45N27 VLP alone and that of HPV18N65 VLP alone. This showed that H18N65-45T3-59S1 VLP, H18N65-45T4-59S1 VLP and H18N65-45T1T3-59S5 VLP had good cross-immunogenicity and cross-protection against HPV18, HPV45 and HPV59.

Example 6: Evaluation 2 of Neutralizing Antibody Titer in Serum of Mice Vaccinated with Virus-Like Particles

(121) In this experiment, virus-like particle used was H18N65-45T4 VLP.

(122) In this experiment, vaccination schedule was shown in Table 13. All the mice (6-week old BalB/c female mice) were divided into 2 groups: Group of dose of 10 μg (at an immunizing dose of 10 μg, using aluminum adjuvant), and Group of dose of 1 μg (at an immunizing dose of 1 μg, using aluminum adjuvant). Each group was further divided into 5 subgroups. The Control subgroups 1 and 2 were vaccinated with HPV45N27 VLP alone and HPV18N65 VLP alone, respectively, the Control subgroup 3 was vaccinated with the mixed HPV18/HPV45 VLP (i.e. a mixture of HPV18N65 VLP and HPV45N27 VLP, at a given immunizing dose for each VLP), and the Experimental subgroup was vaccinated with H18N65-45T4 VLP.

(123) 6 Mice/subgroup were vaccinated by intraperitoneal injection, at an immunizing dose of 10 μg, and 1 μg, respectively, and an injection volume of 1 mL. All the mice were subjected to the first vaccination at Week 0, and then subjected to the booster vaccination at Weeks 2 and 4, respectively. At Week 8, blood sample was collected via orbital bleeding, and the titers of antibodies against HPV18 and HPV45 in serum were analyzed. The analysis results were shown in FIGS. 8A-8B. The result showed that H18N65-45T4 VLP could induce the generation of high-titer neutralizing antibodies against HPV18 in mice, and its protective effect was comparable to that of HPV18N65 VLP alone and that of the mixed HPV18/HPV45 VLP at the same dose, and was significantly superior to that of HPV45N27 VLP alone at the same dose; and it could induce the generation of high-titer neutralizing antibodies against HPV45 in mice, and its protective effect was comparable to that of HPV45N27 VLP alone and that of the mixed HPV18/HPV45 VLP at the same dose, and was significantly superior to that of HPV18N65 VLP alone at the same dose. This showed that H18N65-45T4 VLP had good cross-immunogenicity and cross-protection against HPV18 and HPV45.

(124) TABLE-US-00065 TABLE 13 Vaccination schedule Vaccination Antigen for Immunizing procedure Group vaccination Adjuvant dose Number (week) Group of dose HPV18N65 VLP aluminum 10 μg 6 0, 2, 4 of 10 μg adjuvant HPV45N27 VLP aluminum 10 μg 6 0, 2, 4 adjuvant HPV18/HPV45 VLP aluminum 10 μg for 6 0, 2, 4 adjuvant each VLP H18N65-45T4 VLP aluminum 10 μg 6 0, 2, 4 adjuvant Group of dose HPV18N65 VLP aluminum  1 μg 6 0, 2, 4 of 1 μg adjuvant HPV45N27 VLP aluminum  1 μg 6 0, 2, 4 adjuvant HPV18/HPV45 VLP aluminum  1 μg for 6 0, 2, 4 adjuvant each VLP H18N65-45T4 VLP aluminum  1 μg 6 0, 2, 4 adjuvant

Example 7: Evaluation 3 of Neutralizing Antibody Titer in Serum of Mice Vaccinated with Virus-Like Particles

(125) In this experiment, virus-like particles used were H18N65-45T3-59S1 VLP, H18N65-45T4-59S1 VLP, H18N65-45T1-59S5 VLP, H18N65-45T2-59S5 VLP and H18N65-45T1T3-59S5 VLP.

(126) In this experiment, vaccination schedule was shown in Table 14. All the mice (6-week old BalB/c female mice) were divided into 2 groups: Group of dose of 10 μg (at an immunizing dose of 10 μg, using aluminum adjuvant), and Group of dose of 1 μg (at an immunizing dose of 1 μg, using aluminum adjuvant). Each group was further divided into 9 subgroups. The Control subgroups 1, 2 and 3 were vaccinated with HPV18N65 VLP alone, HPV45N27 VLP alone and HPV59 VLP alone, respectively, the Control subgroup 4 was vaccinated with the mixed HPV18/HPV45/HPV59 VLP (i.e. a mixture of HPV18N65 VLP, HPV45N27 VLP and HPV59 VLP, at a given immunizing dose for each VLP), and the Experimental subgroups 1, 2, 3, 4 and 5 were vaccinated with H18N65-45T3-59S1 VLP, H18N65-45T4-59S1 VLP, H18N65-45T1-59S5 VLP, H18N65-45T2-59S5 VLP and H18N65-45T1T3-59S5 VLP, respectively.

(127) 6 Mice/subgroup were vaccinated by intraperitoneal injection, at an immunizing dose of 10 μg and 1 μg, respectively, and an injection volume of 1 mL. All the mice were subjected to the first vaccination at Week 0, and then subjected to the booster vaccination at Weeks 2 and 4, respectively. At Week 8, blood sample was collected via orbital bleeding, and the titers of antibodies against HPV18, HPV45 and HPV59 in serum were analyzed. The analysis results were shown in FIGS. 8C-8D. The result showed that H18N65-45T3-59S1 VLP, H18N65-45T4-59S1 VLP, H18N65-45T1-59S5 VLP, H18N65-45T2-59S5 VLP and H18N65-45T1T3-59S5 VLP could induce the generation of high-titer neutralizing antibodies against HPV18 in mice, and their protective effects were comparable to that of HPV18N65 VLP alone and that of the mixed HPV18/HPV45/HPV59 VLP at the same dose, and were significantly superior to that of HPV45N27 VLP alone or that of HPV59 VLP alone at the same dose; and H18N65-45T3-59S1 VLP, H18N65-45T4-59S1 VLP, H18N65-45T1-59S5 VLP, H18N65-45T2-59S5 VLP and H18N65-45T1T3-59S5 VLP could induce the generation of high-titer neutralizing antibodies against HPV45 in mice, and their protective effects were comparable to that of HPV45N27 VLP alone and that of the mixed HPV18/HPV45/HPV59 VLP at the same dose, and were significantly superior to that of HPV18N65 VLP alone or that of HPV59 VLP alone at the same dose; and H18N65-45T3-59S1 VLP, H18N65-45T4-59S1 VLP, H18N65-45T1-59S5 VLP, H18N65-45T2-59S5 VLP and H18N65-45T1T3-59S5 VLP could induce the generation of high-titer neutralizing antibodies against HPV59 in mice, and their protective effects were comparable to that of HPV59 VLP alone or that of the mixed HPV18/HPV45/HPV59 VLP at the same dose, and were significantly superior to that of HPV18N65 VLP alone or that of HPV45N27 VLP alone at the same dose. This showed that H18N65-45T3-59S1 VLP, H18N65-45T4-59S1 VLP, H18N65-45T1-59S5 VLP, H18N65-45T2-59S5 VLP and H18N65-45T1T3-59S5 VLP had good cross-immunogenicity and cross-protection against HPV18, HPV45 and HPV59.

(128) TABLE-US-00066 TABLE 14 Vaccination schedule Vaccination Immunizing procedure Group Antigen for vaccination Adjuvant dose Number (week) Group of dose HPV18N65 VLP aluminum 10 μg 6 0, 2, 4 of 10 μg adjuvant HPV45N27 VLP aluminum 10 μg 6 0, 2, 4 adjuvant HPV59 VLP aluminum 10 μg 6 0, 2, 4 adjuvant HPV18/HPV45/HPV59 VLP aluminum 10 μg for 6 0, 2, 4 adjuvant each VLP H18N65-45T3-59S1 VLP aluminum 10 μg 6 0, 2, 4 adjuvant H18N65-45T4-59S1 VLP aluminum 10 μg 6 0, 2, 4 adjuvant H18N65-45T1-59S5 VLP aluminum 10 μg 6 0, 2, 4 adjuvant H18N65-45T2-59S5 VLP aluminum 10 μg 6 0, 2, 4 adjuvant H18N65-45T1T3-59S5 VLP aluminum 10 μg 6 0, 2, 4 adjuvant Group of dose HPV18N65 VLP aluminum  1 μg 6 0, 2, 4 of 1 μg adjuvant HPV45N27 VLP aluminum  1 μg 6 0, 2, 4 adjuvant HPV59 VLP aluminum  1 μg 6 0, 2, 4 adjuvant HPV18/HPV45/HPV59 VLP aluminum  1 μg for 6 0, 2, 4 adjuvant each VLP H18N65-45T3-59S1 VLP aluminum  1 μg 6 0, 2, 4 adjuvant H18N65-45T4-59S1 VLP aluminum  1 μg 6 0, 2, 4 adjuvant H18N65-45T1-59S5 VLP aluminum  1 μg 6 0, 2, 4 adjuvant H18N65-45T2-59S5 VLP aluminum  1 μg 6 0, 2, 4 adjuvant H18N65-45T1T3-59S5 VLP aluminum  1 μg 6 0, 2, 4 adjuvant

(129) Although the specific embodiments of the present invention have been described in details, those skilled in the art would understand that, according to the teachings disclosed in the specification, various modifications and changes can be made thereto, and that such modifications and changes are within the scope of the present invention. The scope of the present invention is given by the appended claims and any equivalents thereof