Vaccines and methods for the treatment of neisseria gonorrhoeae infections

Abstract

Disclosed are novel vaccine compositions comprising multiple N. gonorrhoeae TbpB proteins or immunogenically equivalent portions thereof. The vaccine compositions may be used to ameliorate or prevent pathogenic infections in humans, caused by N. gonorrhoeae. Related methods and uses are also disclosed.

Description

TITLE: VACCINES AND METHODS FOR THE TREATMENT OF NEISSERIA GONORRHOEAE INFECTIONSRELATED APPLICATION

[0001] This application claims the benefit of United States Provisional Application No. 63 / 691 ,519 filed September 6, 2024; the entire contents of Patent Application No. 63 / 691 ,519 are hereby incorporated by reference.GOVERNMENT SUPPORT

[0002] This invention was made with government support under NIH (USA) - Research Project Grant (# 1 R01 Al 141229) awarded by the National Institutes of Health. The U.S. government has certain rights in the invention.FIELD OF THE DISCLOSURE

[0003] The methods, uses, and compositions disclosed herein relate to the treatment of infectious diseases. In particular, the methods, uses, and compositions disclosed herein relate to vaccines to prevent or ameliorate infections in humans caused by the infectious Gram-negative bacterium Neisseria gonorrhoeae.BACKGROUND OF THE DISCLOSURE

[0004] The following paragraphs are provided by way of background to the present disclosure. They are not however an admission that anything discussed therein is prior art or part of the knowledge of persons skilled in the art.

[0005] Gonorrhea is a well-known sexually transmitted infection caused by the bacterium Neisseria gonorrhoeae (N. gonorrhoeae). This bacterium can colonize urogenital, anal and oropharyngeal tissues in humans. It is transmitted during sexual contact, specifically by direct contact between the exposed mucosa lining in the urogenital tract, anal canal, or the pharynx. Disease symptoms vary depending on gender and infection site and can include the development of abnormal penile discharges in men, abnormal vaginal discharges in women, and dysuria (painful urination) in both men and women. If left untreated, gonorrhea can result in infertility in both men and women, and other sexual health complications. According to the World Health Organization (WHO), there were an estimated1745185082.4 million new infections with / V. gonorrhoeae globally among adults in 2020, with incidence rates of 19 (11 - 29) per 1 ,000 women, and 23 (10 - 43) per 1 ,000 men (WHO, Fact Sheet, July 2024, https: / / www.who.int / news-room / fact- sheets / detail / multi-d rug-resistant-gonorrhoea).

[0006] N. gonorrhoeae species can be categorized into N. gonorrhoeae strains, with severity and symptoms of the disease varying depending on the specific infecting strain involved. In order to differentiate between N. gonorrhoeae strains, various typing methods have been developed. These methods fall into two main categories: phenotyping and genotyping. Phenotyping techniques focus on observable characteristics of the strain, for example, differences in nutritional requirements (auxotyping). By contrast, genotyping methods involve the analysis of the genetic material (DNA, RNA) of individual strains. Examples of genotyping techniques include phylogenetic typing, which reconstructs the evolutionary history of N. gonorrhoeae strains by examining DNA sequence variations, and allows classification of related strains into multiple separate clusters of strains.

[0007] Treatment for gonorrhea infections typically involves antibiotic therapies. However, some pathogenic N. gonorrhoeae strains have developed resistance to common front-line antibiotics, including cephalosporins and fluoroquinolones, for example. This growing antibiotic resistance, and resulting treatment failure, is becoming a major global health concern (Unemo M, Golparian D, Eyre DW. Antimicrobial Resistance in Neisseria gonorrhoeae and Treatment of Gonorrhea. Methods Mol Biol. 2019(1940-6029 :37-58). In the United States, the Center for Disease Control (CDC), has identified the public health threat level posed by drug-resistant N. gonorrhoeae as “urgent” (CDC. Antibiotic Resistance Threats in the United States, 2019. Atlanta, GA: U.S. Department of Health and Human Services, CDC; 2019).

[0008] In view of the risks and limitations of currrent antibiotic treatments, there have been suggestions to develop vaccine-based gonorrhea treatments. However, it is not clear whether and how an efficacious vaccine to treat and / or prevent gonorrhea can be developed, and to date no vaccines preventing gonorrhea infections have been approved (Cohen M. et al., 2024, J. Infectious Diseases; https: / / doi.Org / 10.1093 / i nfd i s / j iae 160).

[0009] In light of the foregoing, there exists a need in the art for improved methods and compositions to treat and prevent N. gonorrhoeae infections. In27451850particular, there is a need in the art for vaccines to prevent disease caused by / V. gonorrhoeae infections in humans.SUMMARY OF THE DISCLOSURE

[0010] The present disclosure relates to vaccine formulations and methods of using and making the same.

[0011] In one aspect, the present disclosure relates to vaccine formulations to prevent, treat or ameliorate diseases in humans that are caused by the bacterial pathogen Neisseria gonorrhoeae (N. gonorrhoeae).

[0012] The inventors have discovered that the vaccine formulations of the present disclosure comprising only two immunogenic active agents, can be used to treat infections caused by multiple N. gonorrhoeae strains, including in a broad cross-protective manner. The immunogenic agents included in the vaccine formulations of the present disclosure are, in particular, proteins selected from a class of N. gonorrhoeae proteins, known as TbpB proteins.

[0013] Accordingly, the present disclosure provides, in an aspect, in accordance with the teachings herein, in at least one embodiment, a vaccine formulation for the prevention, treatment, or amelioration of N. gonorrhoeae infection in a human, the vaccine formulation comprising an effective amount of (i) at least two N. gonorrhoeae TbpB proteins, or immunogenically equivalent portions thereof, or (ii) at least two mRNA polynucleotides encoding the two N. gonorrhoeae TbpB proteins, or immunogenically equivalent portions thereof, the at least two N. gonorrhoeae TbpB proteins being selected from at least one Ngo1 TbpB protein, and at least one Ngo2 TbpB protein.

[0014] In at least one embodiment, in an aspect, the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, can be a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 1 , SEQ.ID NO: 3, SEQ.ID NO: 5, SEQ.ID NO: 7, SEQ.ID NO: 19, SEQ.ID NO: 21 , SEQ.ID NO: 23, SEQ.ID NO: 25, SEQ.ID NO: 27, SEQ.ID NO: 29, SEQ.ID NO: 61 , SEQ.ID NO: 63, SEQ.ID NO: 65, SEQ.ID NO: 67, SEQ.ID NO: 69, SEQ.ID NO: 71 , SEQ.ID NO: 73, SEQ.ID NO: 75, SEQ.ID NO: 77, SEQ.ID NO: 79, SEQ.ID NO: 81 , SEQ.ID NO: 83, SEQ.ID37451850NO: 85, SEQ.ID NO: 87, SEQ.ID NO: 101 , SEQ.ID NO: 103, SEQ.ID NO: 105, SEQ.ID NO: 107, SEQ.ID NO: 109, SEQ.ID NO: 111 , SEQ.ID NO: 113, SEQ.ID NO: 115, SEQ.ID NO: 117, SEQ.ID NO: 119, SEQ.ID NO: 121 , SEQ.ID NO: 123, SEQ.ID NO: 125, or SEQ.ID NO: 127;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 2, SEQ.ID NO: 4, SEQ.ID NO: 6, SEQ.ID. NO: 8, SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, SEQ.ID NO: 26, SEQ.ID NO: 28, SEQ.ID NO: 30, SEQ.ID NO: 62, SEQ.ID NO: 64, SEQ.ID NO: 66, SEQ.ID NO: 68, SEQ.ID NO: 70, SEQ.ID NO: 72, SEQ.ID NO: 74, SEQ.ID NO: 76, SEQ.ID NO: 78, SEQ.ID NO: 80, SEQ.ID NO: 82, SEQ.ID NO: 84, SEQ.ID NO: 86, SEQ.ID NO: 88, SEQ.ID NO: 102, SEQ.ID NO: 104, SEQ.ID NO: 106, SEQ.ID NO: 108, SEQ.ID NO: 110, SEQ.ID NO: 112 SEQ.ID NO: 114, SEQ.ID NO: 116, SEQ.ID NO: 187, SEQ.ID NO: 120, SEQ.ID NO: 122, SEQ.ID NO: 124, SEQ.ID NO: 126, or SEQ.ID NO: 128, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 2, SEQ.ID NO: 4, SEQ.ID NO: 6, SEQ.ID. NO: 8, SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, SEQ.ID NO: 26, SEQ.ID NO: 28, SEQ.ID NO: 30, SEQ.ID NO: 62, SEQ.ID NO: 64, SEQ.ID NO: 66, SEQ.ID NO: 68, SEQ.ID NO: 70, SEQ.ID NO: 72, SEQ.ID NO: 74, SEQ.ID NO: 76, SEQ.ID NO: 78, SEQ.ID NO: 80, SEQ.ID NO: 82, SEQ.ID NO: 84, SEQ.ID NO: 86, SEQ.ID NO: 88, SEQ.ID NO: 102, SEQ.ID NO: 104, SEQ.ID NO: 106, SEQ.ID NO: 108, SEQ.ID NO: 110, SEQ.ID NO: 112, SEQ.ID NO: 114, SEQ.ID NO: 116, SEQ.ID NO: 187, SEQ.ID NO: 120, SEQ.ID NO: 122, SEQ.ID NO: 124, SEQ.ID NO: 126, or SEQ.ID NO: 128, or an immunogenically equivalent portion thereto; and47451850(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0015] In at least one embodiment, in an aspect, the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, can be a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 25;(b) a nucleic acid sequence having at least 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with the nucleic acid sequence of (a);(c) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to the nucleic acid sequence of (a);(e) a nucleic acid sequence encoding a polypeptide having the amino acid sequences set forth in SEQ.ID NO: 26, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of the amino acid sequence set forth in SEQ.ID NO: 26, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0016] In at least one embodiment, in an aspect, the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, can be a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 9, SEQ.ID NO: 11 , SEQ.ID NO: 13, SEQ.ID NO: 15, SEQ.ID NO: 17, SEQ.ID NO: 31 , SEQ.ID NO: 32, SEQ.ID NO: 34, SEQ.ID NO: 36, SEQ.ID NO: 37, SEQ.ID NO: 39, SEQ.ID NO: 41 , SEQ.ID NO: 42, SEQ.ID NO: 44, SEQ.ID NO: 46, SEQ.ID NO: 47, SEQ.ID NO: 49, SEQ.ID NO: 51 , SEQ.ID NO: 52, SEQ.ID NO: 54, SEQ.ID NO: 56, SEQ.ID NO: 57, SEQ.ID NO: 59, SEQ.ID NO: 89, SEQ.ID NO: 91 , SEQ.ID NO: 93, SEQ.ID NO: 95, SEQ.ID NO: 97, SEQ.ID NO: 99, SEQ.ID NO: 207, SEQ.ID NO: 209, SEQ.ID NO: 211 , SEQ.ID NO: 213, SEQ.ID NO: 215, SEQ.ID NO: 217, SEQ.ID NO: 219, SEQ.ID NO: 221 , SEQ.ID NO: 223, SEQ.ID NO: 225, SEQ.ID NO: 227,57451850SEQ.ID NO: 229, SEQ.ID NO: 231 , SEQ.ID NO: 233, SEQ.ID NO: 235SEQ.ID NO: 237, SEQ.ID NO: 239, SEQ.ID NO: 241 , SEQ.ID NO: 243SEQ.ID NO: 245, SEQ.ID NO: 247, SEQ.ID NO: 249, SEQ.ID NO: 251SEQ.ID NO: 253, SEQ.ID NO: 255, SEQ.ID NO: 257, SEQ.ID NO: 259SEQ.ID NO: 261 , SEQ.ID NO: 263, SEQ.ID NO: 265, SEQ.ID NO: 267SEQ.ID NO: 269, SEQ.ID NO: 271 , SEQ.ID NO: 273, SEQ.ID NO: 343, orSEQ.ID NO: 345;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.ID NO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45,SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.IDNO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94,SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.IDNO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO: 216, SEQ.ID NO:218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224, SEQ.ID NO: 226,SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232, SEQ.ID NO: 234SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240, SEQ.ID NO: 242SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248, SEQ.ID NO: 250SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256, SEQ.ID NO: 258SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264, SEQ.ID NO: 266SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272, SEQ.ID NO: 274SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.ID67451850NO: 35, SEQ.ID NO: 38, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.IDNO: 55, SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO: 216, SEQ.ID NO: 218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224,SEQ.ID NO: 226, SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232,SEQ.ID NO: 234, SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240,SEQ.ID NO: 242, SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248,SEQ.ID NO: 250, SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256,SEQ.ID NO: 258, SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264,SEQ.ID NO: 266, SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272,SEQ.ID NO: 27. , SEQ.ID NO: >44, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0017] In at least one embodiment, in an aspect, the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, can be a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 1 , SEQ.ID NO: 3, SEQ.ID NO: 5, SEQ.ID NO: 7, SEQ.ID NO: 19, SEQ.ID NO: 21 , SEQ.ID NO: 23, SEQ.ID NO: 25, SEQ.ID NO: 27, SEQ.ID NO: 29, SEQ.ID NO: 61 , SEQ.ID NO: 63, SEQ.ID NO: 65, SEQ.ID NO: 67, SEQ.ID NO: 69, SEQ.ID NO: 71 , SEQ.ID NO: 73, SEQ.ID NO: 75, SEQ.ID NO: 77, SEQ.ID NO: 79, SEQ.ID NO: 81 , SEQ.ID NO: 83, SEQ.ID NO: 85, SEQ.ID NO: 87, SEQ.ID NO: 101 , SEQ.ID NO: 103, SEQ.ID NO:105, SEQ.ID NO: 107, SEQ.ID NO: 109, SEQ.ID NO: 111 , SEQ.ID NO: 113, SEQ.ID NO: 115, SEQ.ID NO: 117, SEQ.ID NO: 119, SEQ.ID NO: 121 , SEQ.ID NO: 123, SEQ.ID NO: 125, or SEQ.ID NO: 127;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;77451850(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 2, SEQ.ID NO: 4, SEQ.ID NO: 6, SEQ.ID NO: 8, SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, SEQ.ID NO: 26, SEQ.ID NO: 28, SEQ.ID NO: 30, SEQ.ID NO: 62, SEQ.ID NO: 64, SEQ.ID NO: 66, SEQ.ID NO: 68, SEQ.ID NO: 70, SEQ.ID NO: 72, SEQ.ID NO: 74, SEQ.ID NO: 76, SEQ.ID NO: 78, SEQ.ID NO: 80, SEQ.ID NO: 82, SEQ.ID NO: 84, SEQ.ID NO: 86, SEQ.ID NO: 88, SEQ.ID NO: 102, SEQ.ID NO: 104, SEQ.ID NO: 106, SEQ.ID NO: 108, SEQ.ID NO: 110, SEQ.ID NO: 112, SEQ.ID NO: 114, SEQ.ID NO: 116, SEQ.ID NO: 187, SEQ.ID NO: 120, SEQ.ID NO: 122, SEQ.ID NO: 124, SEQ.ID NO: 126, or SEQ.ID NO: 128, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 2, SEQ.ID NO: 4, SEQ.ID NO: 6, SEQ.ID NO: 8, SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, SEQ.ID NO: 26, SEQ.ID NO: 28, SEQ.ID NO: 30, SEQ.ID NO: 62, SEQ.ID NO: 64, SEQ.ID NO: 66, SEQ.ID NO: 68, SEQ.ID NO: 70, SEQ.ID NO: 72, SEQ.ID NO: 74, SEQ.ID NO: 76, SEQ.ID NO: 78, SEQ.ID NO: 80, SEQ.ID NO: 82, SEQ.ID NO: 84, SEQ.ID NO: 86, SEQ.ID NO: 88, SEQ.ID NO: 102, SEQ.ID NO: 104, SEQ.ID NO: 106, SEQ.ID NO: 108, SEQ.ID NO: 110, SEQ.ID NO: 112, SEQ.ID NO: 114, SEQ.ID NO: 116, SEQ.ID NO: 187, SEQ.ID NO: 120, SEQ.ID NO: 122, SEQ.ID NO: 124, SEQ.ID NO: 126, or SEQ.ID NO: 128, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f); and the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, can be a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 9, SEQ.ID NO: 11 , SEQ.ID NO: 13, SEQ.ID NO: 15, SEQ.ID NO: 17, SEQ.ID NO: 31 , SEQ.ID NO: 32, SEQ.ID NO: 34, SEQ.ID NO: 36, SEQ.ID NO: 37, SEQ.ID NO: 39, SEQ.ID NO: 41 , SEQ.ID NO: 42, SEQ.ID NO: 44, SEQ.ID NO: 46, SEQ.ID NO: 47, SEQ.ID NO: 49, SEQ.ID NO: 51 ,87451850SEQ.ID NO: 52, SEQ.ID NO: 54, SEQ.ID NO: 56, SEQ.ID NO: 57, SEQ.ID NO: 59, SEQ.ID NO: 89, SEQ.ID NO: 91 , SEQ.ID NO: 93, SEQ.ID NO: 95,SEQ.ID NO: 97, SEQ.ID NO: 99, SEQ.ID NO: 207, SEQ.ID NO: 209, SEQ.IDNO: 211 , SEQ.ID NO: 213, SEQ.ID NO: 215, SEQ.ID NO: 217, SEQ.ID NO:219, SEQ.ID NO: 221 , SEQ.ID NO: 223, SEQ.ID NO: 225, SEQ.ID NO: 227,SEQ.ID NO: 229, SEQ.ID NO: 231 , SEQ.ID NO: 233, SEQ.ID NO: 235SEQ.ID NO: 237, SEQ.ID NO: 239, SEQ.ID NO: 241 , SEQ.ID NO: 243SEQ.ID NO: 245, SEQ.ID NO: 247, SEQ.ID NO: 249, SEQ.ID NO: 251SEQ.ID NO: 253, SEQ.ID NO: 255, SEQ.ID NO: 257, SEQ.ID NO: 259SEQ.ID NO: 261 , SEQ.ID NO: 263, SEQ.ID NO: 265, SEQ.ID NO: 267SEQ.ID NO: 269, SEQ.ID NO: 271 , SEQ.ID NO: 273, SEQ.ID NO: 343, or SEQ.ID NO: 345;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.ID NO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.IDNO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO: 216, SEQ.ID NO218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224, SEQ.ID NO: 226SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232, SEQ.ID NO: 234,SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240, SEQ.ID NO: 242,SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248, SEQ.ID NO: 250,SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256, SEQ.ID NO: 258,SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264, SEQ.ID NO: 266,SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272, SEQ.ID NO: 274,97451850SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.ID NO: 35, SEQ.ID NO: 38, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO:216, SEQ.ID NO: 218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224,SEQ.ID NO: 226, SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232,SEQ.ID NO: 234, SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240,SEQ.ID NO: 242, SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248,SEQ.ID NO: 250, SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256,SEQ.ID NO: 258, SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264,SEQ.ID NO: 266, SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272,SEQ.ID NO: 274, SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0018] In at least one embodiment, in an aspect, the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, can be a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 19, SEQ.ID NO: 21 , SEQ.ID NO: 23, orSEQ.ID NO: 25;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);107451850(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, orSEQ.ID NO: 26, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, orSEQ.ID NO: 26, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f); and the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, can be a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 9, SEQ.ID NO: 11 , SEQ.ID NO: 13, SEQ.ID NO: 15, SEQ.ID NO: 17, SEQ.ID NO: 31 , SEQ.ID NO: 32, SEQ.ID NO: 34, SEQ.ID NO: 36, SEQ.ID NO: 37, SEQ.ID NO: 39, SEQ.ID NO: 41 , SEQ.ID NO: 42, SEQ.ID NO: 44 SEQ.ID NO: 46, SEQ.ID NO: 47, SEQ.ID NO: 49, SEQ.ID NO: 51 ,SEQ.ID NO: 52, SEQ.ID NO: 54, SEQ.ID NO: 56, SEQ.ID NO: 57, SEQ.IDNO: 59, SEQ.ID NO: 89, SEQ.ID NO: 91 , SEQ.ID NO: 93, SEQ.ID NO: 95,SEQ.ID NO: 97, SEQ.ID NO: 99, SEQ.ID NO: 207, SEQ.ID NO: 209, SEQ.IDNO: 211 , SEQ.ID NO: 213, SEQ.ID NO: 215, SEQ.ID NO: 217, SEQ.ID NO:219, SEQ.ID NO: 221 , SEQ.ID NO: 223, SEQ.ID NO: 225, SEQ.ID NO: 227,SEQ.ID NO: 229, SEQ.ID NO: 231 , SEQ.ID NO: 233, SEQ.ID NO: 235,SEQ.ID NO: 237, SEQ.ID NO: 239, SEQ.ID NO: 241 , SEQ.ID NO: 243,SEQ.ID NO: 245, SEQ.ID NO: 247, SEQ.ID NO: 249, SEQ.ID NO: 251 ,SEQ.ID NO: 253, SEQ.ID NO: 255, SEQ.ID NO: 257, SEQ.ID NO: 259,SEQ.ID NO: 261 , SEQ.ID NO: 263, SEQ.ID NO: 265, SEQ.ID NO: 267,SEQ.ID NO: 269, SEQ.ID NO: 271 , SEQ.ID NO: 273, SEQ.ID NO: 343, or SEQ.ID NO: 345;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);117451850(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.ID NO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.IDNO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94,SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.IDNO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO: 216, SEQ.ID NO:218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224, SEQ.ID NO: 226,SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232, SEQ.ID NO: 234,SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240, SEQ.ID NO: 242,SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248, SEQ.ID NO: 250,SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256, SEQ.ID NO: 258,SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264, SEQ.ID NO: 266,SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272, SEQ.ID NO: 274, SEQ.ID NO: 344, or SEQ.ID NO: 3< 6, or an immunoc enically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.IDNO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45,SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.IDNO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94,SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.IDNO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO: 216, SEQ.ID NO:218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224, SEQ.ID NO: 226,SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232, SEQ.ID NO: 234,SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240, SEQ.ID NO: 242,SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248, SEQ.ID NO: 250,127451850SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256, SEQ.ID NO: 258,SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264, SEQ.ID NO: 266,SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272, SEQ.ID NO: 274,SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0019] In at least one embodiment, in an aspect, the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, can be a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 19, SEQ.ID NO: 21 , SEQ.ID NO: 23, orSEQ.ID NO: 25;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, orSEQ.ID NO: 26, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, orSEQ.ID NO: 26, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f); and the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, can be a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:137451850(a) SEQ.ID NO: 9, SEQ.ID NO: 11 , SEQ.ID NO: 13, SEQ.ID NO: 15, SEQ.ID NO: 17, SEQ.ID NO: 89, SEQ.ID NO: 91 , SEQ.ID NO: 93, SEQ.ID NO: 95, SEQ.ID NO: 97, or SEQ.ID NO: 99;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, or SEQ.ID NO: 100, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, or SEQ.ID NO: 100, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0020] In at least one embodiment, in an aspect, the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, can be a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 25;(b) a nucleic acid sequence having at least 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with the nucleic acid sequence of (a);(c) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to the nucleic acid sequence of (a);147451850(e) a nucleic acid sequence encoding a polypeptide having the amino acid sequences set forth in SEQ.ID NO: 26, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of the amino acid sequence set forth in SEQ.ID NO: 26, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f); and the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, can be a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 9, SEQ.ID NO: 11 , SEQ.ID NO: 13, SEQ.ID NO: 15, SEQ.ID NO: 17, SEQ.ID NO: 31 , SEQ.ID NO: 32, SEQ.ID NO: 34, SEQ.ID NO: 36, SEQ.ID NO: 37, SEQ.ID NO: 39, SEQ.ID NO: 41 , SEQ.ID NO: 42, SEQ.ID NO: 44, SEQ.ID NO: 46, SEQ.ID NO: 47, SEQ.ID NO: 49, SEQ.ID NO: 51 , SEQ.ID NO: 52, SEQ.ID NO: 54, SEQ.ID NO: 56, SEQ.ID NO: 57, SEQ.ID NO: 59, SEQ.ID NO: 89, SEQ.ID NO: 91 , SEQ.ID NO: 93, SEQ.ID NO: 95,SEQ.ID NO: 97, SEQ.ID NO: 99, SEQ.ID NO: 207, SEQ.ID NO: 209, SEQ.IDNO: 211 , SEQ.ID NO: 213, SEQ.ID NO: 215, SEQ.ID NO: 217, SEQ.ID NO:219, SEQ.ID NO: 221 , SEQ.ID NO: 223, SEQ.ID NO: 225, SEQ.ID NO: 227,SEQ.ID NO: 229, SEQ.ID NO: 231 , SEQ.ID NO: 233, SEQ.ID NO: 235SEQ.ID NO: 237, SEQ.ID NO: 239, SEQ.ID NO: 241 , SEQ.ID NO: 243SEQ.ID NO: 245, SEQ.ID NO: 247, SEQ.ID NO: 249, SEQ.ID NO: 251SEQ.ID NO: 253, SEQ.ID NO: 255, SEQ.ID NO: 257, SEQ.ID NO: 259SEQ.ID NO: 261 , SEQ.ID NO: 263, SEQ.ID NO: 265, SEQ.ID NO: 267SEQ.ID NO: 269, SEQ.ID NO: 271 , SEQ.ID NO: 273, SEQ.ID NO: 343, or SEQ.ID NO: 345;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;157451850(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.ID NO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO: 216, SEQ.ID NO:218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224, SEQ.ID NO: 226,SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232, SEQ.ID NO: 234,SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240, SEQ.ID NO: 242,SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248, SEQ.ID NO: 250,SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256, SEQ.ID NO: 258,SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264, SEQ.ID NO: 266,SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272, SEQ.ID NO: 274, SEQ.ID NO: 344, or SEQ.ID NO: 3< 6, or an immunoc enically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.IDNO: 35, SEQ.ID NO: 38, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.IDNO: 208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO216, SEQ.ID NO: 218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224SEQ.ID NO: 226, SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232,SEQ.ID NO: 234, SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240,SEQ.ID NO: 242, SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248,SEQ.ID NO: 250, SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256,SEQ.ID NO: 258, SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264,SEQ.ID NO: 266, SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272,167451850SEQ.ID NO: 274, SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0021] In at least one embodiment, in an aspect, the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, can be a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 25;(b) a nucleic acid sequence having at least 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with the nucleic acid sequence of (a);(c) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to the nucleic acid sequence of (a);(e) a nucleic acid sequence encoding a polypeptide having the amino acid sequences set forth in SEQ.ID NO: 26, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of the amino acid sequence set forth in SEQ.ID NO: 26, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f); and the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, can be a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 15, or SEQ.ID NO: 17;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;177451850(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 16 or SEQ.ID NO: 18, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 16 or SEQ.ID NO: 18, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0022] In at least one embodiment, in an aspect, the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, can comprise any of the amino acid sequences set forth in: SEQ.ID NO: 2, SEQ.ID NO: 4, SEQ.ID NO: 6, SEQ.ID NO: 8, SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, SEQ.ID NO: 26, SEQ.ID NO: 28, SEQ.ID NO: 30, SEQ.ID NO: 62, SEQ.ID NO: 64, SEQ.ID NO: 66, SEQ.ID NO: 68, SEQ.ID NO: 70, SEQ.ID NO: 72, SEQ.ID NO: 74, SEQ.ID NO: 76, SEQ.ID NO: 78, SEQ.ID NO: 80, SEQ.ID NO: 82, SEQ.ID NO: 84, SEQ.ID NO: 86, SEQ.ID NO: 88, SEQ.ID NO: 102, SEQ.ID NO: 104, SEQ.ID NO: 106, SEQ.ID NO: 108, SEQ.ID NO: 110, SEQ.ID NO: 112, SEQ.ID NO: 114, SEQ.ID NO: 116, SEQ.ID NO: 187, SEQ.ID NO: 120, SEQ.ID NO: 122, SEQ.ID NO: 124, SEQ.ID NO: 126, or SEQ.ID NO: 128, or an immunologically equivalent portion thereof.

[0023] In at least one embodiment, in an aspect, the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, can comprise any of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.ID NO: 35, SEQ.IDNO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.IDNO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.IDNO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.IDNO: 100, SEQ.ID NO: 208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO: 216, SEQ.ID NO: 218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224, SEQ.ID NO: 226, SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232, SEQ.ID NO: 234, SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240, SEQ.ID187451850NO: 242, SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248, SEQ.ID NO: 250, SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256, SEQ.ID NO: 258, SEQ.IDNO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264, SEQ.ID NO: 266, SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272, SEQ.ID NO: 274, SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereof.

[0024] In at least one embodiment, in an aspect, the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, can comprise: any of the amino acid sequences set forth in: SEQ.ID NO: 2, SEQ.ID NO: 4, SEQ.ID NO: 6, SEQ.ID NO: 8, SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, SEQ.ID NO: 26, SEQ.ID NO: 28, SEQ.ID NO: 30, SEQ.IDNO: 62, SEQ.ID NO: 64, SEQ.ID NO: 66, SEQ.ID NO: 68, SEQ.ID NO: 70, SEQ.ID NO: 72, SEQ.ID NO: 74, SEQ.ID NO: 76, SEQ.ID NO: 78, SEQ.ID NO: 80, SEQ.ID NO: 82, SEQ.ID NO: 84, SEQ.ID NO: 86, SEQ.ID NO: 88,SEQ.ID NO: 102, SEQ.ID NO: 104, SEQ.ID NO: 106, SEQ.ID NO: 108,SEQ.ID NO: 110, SEQ.ID NO: 112, SEQ.ID NO: 114, SEQ.ID NO: 116, SEQ.ID NO: 187, SEQ.ID NO: 120, SEQ.ID NO: 122, SEQ.ID NO: 124, SEQ.ID NO: 126, or SEQ.ID NO: 128, or an immunologically equivalent portion thereof; and the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, can comprise: any of the amino acid sequences setforth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.ID NO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.IDNO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, or SEQ.ID NO: 100, SEQ.ID NO208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO216, SEQ.ID NO: 218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO224, SEQ.ID NO: 226, SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO232, SEQ.ID NO: 234, SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO240, SEQ.ID NO: 242, SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO248, SEQ.ID NO: 250, SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO256, SEQ.ID NO: 258, SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO197451850264, SEQ.ID NO: 266, SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO:272, SEQ.ID NO: 274, SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereof.

[0025] In at least one embodiment, in an aspect, the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, can comprise:SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, orSEQ.ID NO: 26, or an immunologically equivalent portion thereof; and the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, can comprise: any of the amino acid sequences setforth in SEQ.ID NO: 10, SEQ.IDNO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33,SEQ.ID NO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.IDNO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55,SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.IDNO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO:208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO:216, SEQ.ID NO: 218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO:224, SEQ.ID NO: 226, SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO:232, SEQ.ID NO: 234, SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO:240, SEQ.ID NO: 242, SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO:248, SEQ.ID NO: 250, SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO:256, SEQ.ID NO: 258, SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO:264, SEQ.ID NO: 266, SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO:272, SEQ.ID NO: 274, SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereof.

[0026] In at least one embodiment, in an aspect, the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, can comprise SEQ.ID NO: 26, or an immunologically equivalent portion thereof; and the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, can comprise: any of the amino acid sequences setforth in SEQ.ID NO: 10, SEQ.IDNO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33,207451850SEQ.ID NO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55,SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.IDNO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO:208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO:216, SEQ.ID NO: 218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO:224, SEQ.ID NO: 226, SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO:232, SEQ.ID NO: 234, SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO:240, SEQ.ID NO: 242, SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO:248, SEQ.ID NO: 250, SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO:256, SEQ.ID NO: 258, SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO:264, SEQ.ID NO: 266, SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO:272, SEQ.ID NO: 274, SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereof.

[0027] In at least one embodiment, in an aspect, the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, can comprise:SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, orSEQ.ID NO: 26, or an immunologically equivalent portion thereof; and the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, can comprise: any of the amino acid sequences setforth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, or SEQ.ID NO: 100, or an immunogenically equivalent portion thereof.

[0028] In at least one embodiment, in an aspect, at least one of the at least one Ngo1 TbpB protein and / or the at least one Ngo2 TbpB protein can be a modulated TbpB protein, such that the Kd between the modulated TbpB protein and a native host iron binding protein exceeds the Kd between the native nonmodified TbpB protein and the native host iron binding protein by a factor of at least 2x, at least 10x, or at least 100x, wherein, optionally, the native non-modified TbpB protein and the modulated TbpB protein each comprise at least 100 consecutive amino acid residues which are identical to one another, but for one,217451850two, or three modulated amino acid residues comprised within the at least 100 consecutive amino acid residues of the modulated TbpB protein.

[0029] In at least one embodiment, in an aspect, the modulated TbpB protein can be an Ngo1 protein comprising an amino acid sequence selected from SEQ.ID NO: 26, SEQ.ID NO: 62, SEQ.ID NO: 64, SEQ.ID NO: 68, SEQ.ID NO: 70, SEQ.ID NO: 72, SEQ.ID NO: 74, SEQ.ID NO: 76, SEQ.ID NO:80, SEQ.ID NO: 82, SEQ.ID NO: 84, SEQ.ID NO: 86, SEQ.ID NO: 88, SEQ.ID NO: 102, SEQ.ID NO: 108, SEQ.ID NO: 110, SEQ.ID NO: 116, SEQ.ID NO: 122, and SEQ.ID NO: 128, and the Ngo2 protein can be a modulated TbpB protein comprising an amino acid selected from SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 48, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO: 216, SEQ.ID NO: 218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224, SEQ.ID NO: 226, SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232, SEQ.ID NO: 234, SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240, SEQ.ID NO: 242, SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248, SEQ.ID NO: 250, SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256, SEQ.ID NO: 258, SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264, SEQ.ID NO: 266, SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272, SEQ.ID NO: 274, SEQ.ID NO: 344, and SEQ.ID NO: 346.

[0030] In at least one embodiment, in an aspect, the modulated TbpB protein can be an Ngo1 protein comprising an amino acid sequence selected from SEQ.ID NO: 26, SEQ.ID NO: 102, and SEQ.ID NO: 108, and the Ngo2 protein can be a modulated TbpB protein comprising an amino acid selected from SEQ.ID NO: 230, SEQ.ID NO: 234, SEQ.ID NO: 238, and SEQ.ID NO: 346.

[0031] In at least one embodiment, in an aspect, the native non-modified TbpB protein can be an Ngo1 TbpB protein comprising at least 100 consecutive amino acid residues of SEQ.ID NO: 22.

[0032] In at least one embodiment, in an aspect, the native non-modified TbpB protein can be an Ngo1 TbpB protein comprising at least 100 consecutive amino acid residues of SEQ.ID NO: 22, and the modulated TbpB protein comprises one modulated amino acid residue comprised within the at least 100 consecutive amino acid residues, the modulated amino acid residue corresponding with aspartic acid amino acid residue 209 (209D) in SEQ.ID NO:22745185022, and wherein, optionally, aspartic acid amino acid residue 209 (209D) is modulated to replace aspartic acid amino acid residue 209 (209D) in the modulated TbpB protein with a positively charged amino acid residue, optionally, a histidine (H) amino acid residue.

[0033] In at least one embodiment, in an aspect, the native non-modified protein can be an Ngo1 TbpB protein comprising at least 100 consecutive amino acid residues of SEQ.ID NO: 22, and the modulated TbpB protein comprises SEQ.ID NO: 26.

[0034] In at least one embodiment, in an aspect, the vaccine formulation can comprise a fusion polypeptide comprising the first and second N. gonorrhoeae TbpB protein, or immunologically equivalent portion thereof.

[0035] In at least one embodiment, in an aspect, the vaccine formulation can comprise two N. gonorrhoeae TbpB proteins, or immunogenically equivalent portions thereof.

[0036] In at least one embodiment, in an aspect, the vaccine formulation can comprise three, four, or five, N. gonorrhoeae TbpB proteins, or immunogenically equivalent portions thereof.

[0037] In at least one embodiment, in an aspect, the vaccine formulation can be a cross-protective vaccine formulation, wherein the at least two TbpB proteins, or immunogenically equivalent portions thereof, are obtained from a first and second N. gonorrhoeae strain, and the vaccine formulation is for the administration to a human to prevent or ameliorate an infection caused by a strain other than the first or second N. gonorrhoeae strain.

[0038] In at least one embodiment, the infection can be caused by an N. gonorrhoeae strain selected from MS11 , FA1090, FA19, WHO F, WHO G, WHO L, WHO M, WHO O, WHO, P, WHO V, WHO W, WHO X, WHO Y.

[0039] In at least one embodiment, in an aspect, the at least two TbpB proteins, or immunogenically equivalent portions thereof, can be recombinantly produced in a host cell, optionally a microbial host cell.

[0040] In at least one embodiment, in an aspect, the at least two TbpB proteins, or immunologically equivalent portions thereof, can be recombinantly produced in a host cell, preferably a microbial host cell, wherein the vaccine formulation is substantially free of host cell constituents, other than the at least two TbpB polypeptides, or immunologically equivalent portions thereof.237451850

[0041] In at least one embodiment, in an aspect, the vaccine formulation can be substantially free of N. gonorrhoeae constituents, other than the at least two TbpB polypeptides, or immunologically equivalent portions thereof.

[0042] In at least one embodiment, in an aspect, the vaccine formulation can further comprise a pharmaceutically acceptable adjuvant.

[0043] In at least one embodiment, in an aspect, the vaccine formulation can further comprise a pharmaceutically acceptable excipient, carrier, or diluent.

[0044] In at least one embodiment, in an aspect, the vaccine formulation can comprise from about 0.001 % to about 20% by weight of the at least two TbpB proteins, or the immunogenically equivalent portions thereof, and a pharmaceutically acceptable adjuvant constituting from about 0.1 % to about 60% by weight or volume of the vaccine formulation.

[0045] In another aspect, the present disclosure provides, in accordance with the teachings herein, in at least one embodiment, a method for prevention, treatment, or amelioration of / V. gonorrhoeae infection in a human in need thereof, the method comprising administering to a human a vaccine formulation comprising an effective amount of (i) at least two N. gonorrhoeae TbpB proteins, or immunogenically equivalent portions thereof, or (ii) at least two mRNA polynucleotides encoding the two N. gonorrhoeae TbpB proteins, or immunogenically equivalent portions thereof, the at least two N. gonorrhoeae TbpB proteins being selected from at least one Ngo1 TbpB protein, and at least one Ngo2 TbpB protein, and wherein the vaccine formulation is administered in an effective amount to prevent, treat or ameliorate the N. gonorrhoeae infection.

[0046] In another aspect, the present disclosure provides, in accordance with the teachings herein, in at least one embodiment, a use of a vaccine formulation for the prevention, treatment or amelioration of N. gonorrhoeae infection in a human in need thereof, the vaccine formulation comprising an effective amount of (i) at least two N. gonorrhoeae TbpB proteins, or immunogenically equivalent portions thereof, or (ii) at least two mRNA polynucleotides encoding the two N. gonorrhoeae TbpB proteins, or immunogenically equivalent portions thereof, the at least two N. gonorrhoeae TbpB proteins being selected from at least one Ngo1 TbpB protein, and at least one Ngo2 TbpB protein.247451850

[0047] In another aspect, the present disclosure provides, in at least one embodiment, a use of a vaccine formulation in the manufacture of a medicament for the prevention, treatment or amelioration of N. gonorrhoeae infection in a human in need thereof, the vaccine formulation comprising an effective amount of(i) at least two N. gonorrhoeae TbpB proteins, or immunogenically equivalent portions thereof, or (ii) at least two mRNA polynucleotides encoding the two N. gonorrhoeae TbpB proteins, or immunogenically equivalent portions thereof, the at least two N. gonorrhoeae TbpB proteins being selected from at least one Ngo1 TbpB protein, and at least one Ngo2 TbpB protein.

[0048] In another aspect, the present disclosure provides, in accordance with the teachings herein, in at least one embodiment, a vaccine formulation for use in the prevention, treatment or amelioration of N. gonorrhoeae infection in a human, the vaccine formulation comprising an effective amount of (i) at least two N. gonorrhoeae TbpB proteins, or immunogenically equivalent portions thereof, or(ii) at least two mRNA polynucleotides encoding the two N. gonorrhoeae TbpB proteins, or immunogenically equivalent portions thereof, the at least two N. gonorrhoeae TbpB proteins being selected from at least one Ngo1 TbpB protein, and at least one Ngo2 TbpB protein.

[0049] In at least one embodiment, in an aspect, the treatment or amelioration of the N. gonorrhoeae infection can comprise the reduction of clinical signs of any disease caused by the N. gonorrhoeae infection.

[0050] In at least one embodiment, in an aspect, the at least two TbpB proteins can be TbpB proteins other than from a N. gonorrhoeae strain causing the N. gonorrhoeae infection in the human.

[0051] In at least one embodiment, in an aspect, the clinical signs can be selected from the group consisting of dysuria, pharyngitis, abnormal penile discharge, abnormal vaginal discharge, disseminated gonococcal infection (DGI), including DGI-associated arthritis, tenosynovitis, endocarditis and / or dermatitis, relative to a human infected by N. gonorrhoeae not having been administered the vaccine formulation.

[0052] In at least one embodiment, in an aspect, the vaccine formulation can be a cross-protective vaccine formulation, wherein the at least two TbpB proteins, or immunogenically equivalent portions thereof, are obtained from a first and second N. gonorrhoeae strain, and the vaccine formulation is for the257451850administration to a human to prevent or ameliorate an infection caused by a strain other than the first or second N. gonorrhoeae strain.

[0053] In at least one embodiment, the infection can be caused by an N. gonorrhoeae strain selected from MS11 , FA1090, FA19, WHO F, WHO G, WHO L, WHO M, WHO O, WHO, P, WHO V, WHO W, WHO X, WHO Y.

[0054] In at least one embodiment, in an aspect, the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, can be a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 1 , SEQ.ID NO: 3, SEQ.ID NO: 5, SEQ.ID NO: 7, SEQ.ID NO: 19, SEQ.ID NO: 21 , SEQ.ID NO: 23, SEQ.ID NO: 25, SEQ.ID NO: 27, SEQ.ID NO: 29, SEQ.ID NO: 61 , SEQ.ID NO: 63, SEQ.ID NO: 65, SEQ.ID NO: 67, SEQ.ID NO: 69, SEQ.ID NO: 71 , SEQ.ID NO: 73, SEQ.ID NO: 75, SEQ.ID NO: 77, SEQ.ID NO: 79, SEQ.ID NO: 81 , SEQ.ID NO: 83, SEQ.ID NO: 85, SEQ.ID NO: 87, SEQ.ID NO: 101 , SEQ.ID NO: 103, SEQ.ID NO: 105, SEQ.ID NO: 107, SEQ.ID NO: 109, SEQ.ID NO: 111 , SEQ.ID NO: 113, SEQ.ID NO: 115, SEQ.ID NO: 117, SEQ.ID NO: 119, SEQ.ID NO: 121 , SEQ.ID NO: 123, SEQ.ID NO: 125, or SEQ.ID NO: 127;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 2, SEQ.ID NO: 4, SEQ.ID NO: 6, SEQ.ID. NO: 8, SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, SEQ.ID NO: 26, SEQ.ID NO: 28, SEQ.ID NO: 30, SEQ.ID NO: 62, SEQ.ID NO: 64, SEQ.ID NO: 66, SEQ.ID NO: 68, SEQ.ID NO: 70, SEQ.ID NO: 72, SEQ.ID NO: 74, SEQ.ID NO: 76, SEQ.ID NO: 78, SEQ.ID NO: 80, SEQ.ID NO: 82, SEQ.ID NO: 84, SEQ.ID NO: 86, SEQ.ID NO: 88, SEQ.ID NO: 102, SEQ.ID NO: 104, SEQ.ID NO: 106, SEQ.ID NO: 108, SEQ.ID NO: 110, SEQ.ID NO: 112 SEQ.ID NO: 114, SEQ.ID NO: 116, SEQ.ID NO: 187,267451850SEQ.ID NO: 120, SEQ.ID NO: 122, SEQ.ID NO: 124, SEQ.ID NO: 126, or SEQ.ID NO: 128, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 2, SEQ.ID NO: 4, SEQ.ID NO: 6, SEQ.ID. NO: 8, SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, SEQ.ID NO: 26, SEQ.ID NO: 28, SEQ.ID NO: 30, SEQ.ID NO: 62, SEQ.ID NO: 64, SEQ.ID NO: 66, SEQ.ID NO: 68, SEQ.ID NO: 70, SEQ.ID NO: 72, SEQ.ID NO: 74, SEQ.ID NO: 76, SEQ.ID NO: 78, SEQ.ID NO: 80, SEQ.ID NO: 82, SEQ.ID NO: 84, SEQ.ID NO: 86, SEQ.ID NO: 88, SEQ.ID NO: 102, SEQ.ID NO: 104, SEQ.ID NO: 106, SEQ.ID NO: 108, SEQ.ID NO: 110, SEQ.ID NO: 112 SEQ.ID NO: 114, SEQ.ID NO: 116, SEQ.ID NO: 187, SEQ.ID NO: 120, SEQ.ID NO: 122, SEQ.ID NO: 124, SEQ.ID NO: 126, or SEQ.ID NO: 128, or an immunogenically equivalent portion thereto;(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f); and the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, can be a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 9, SEQ.ID NO: 11 , SEQ.ID NO: 13, SEQ.ID NO: 15, SEQ.ID NO: 17, SEQ.ID NO: 31 , SEQ.ID NO: 32, SEQ.ID NO: 34, SEQ.ID NO: 36, SEQ.ID NO: 37, SEQ.ID NO: 39, SEQ.ID NO: 41 , SEQ.ID NO: 42, SEQ.ID NO: 44, SEQ.ID NO: 46, SEQ.ID NO: 47, SEQ.ID NO: 49, SEQ.ID NO: 51 , SEQ.ID NO: 52, SEQ.ID NO: 54, SEQ.ID NO: 56, SEQ.ID NO: 57, SEQ.ID NO: 59, SEQ.ID NO: 89, SEQ.ID NO: 91 , SEQ.ID NO: 93, SEQ.ID NO: 95, SEQ.ID NO: 97, SEQ.ID NO: 99, SEQ.ID NO: 207, SEQ.ID NO: 209, SEQ.IDNO: 211 , SEQ.ID NO: 213, SEQ.ID NO: 215, SEQ.ID NO: 217, SEQ.ID NO219, SEQ.ID NO: 221 , SEQ.ID NO: 223, SEQ.ID NO: 225, SEQ.ID NO: 227SEQ.ID NO: 229, SEQ.ID NO: 231 , SEQ.ID NO: 233, SEQ.ID NO: 235,SEQ.ID NO: 237, SEQ.ID NO: 239, SEQ.ID NO: 241 , SEQ.ID NO: 243,SEQ.ID NO: 245, SEQ.ID NO: 247, SEQ.ID NO: 249, SEQ.ID NO: 251 ,SEQ.ID NO: 253, SEQ.ID NO: 255, SEQ.ID NO: 257, SEQ.ID NO: 259,SEQ.ID NO: 261 , SEQ.ID NO: 263, SEQ.ID NO: 265, SEQ.ID NO: 267,277451850SEQ.ID NO: 269, SEQ.ID NO: 271 , SEQ.ID NO: 273, SEQ.ID NO: 343, or SEQ.ID NO: 345;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.ID NO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO: 216, SEQ.ID NO: 218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224, SEQ.ID NO: 226,SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232, SEQ.ID NO: 234SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240, SEQ.ID NO: 242SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248, SEQ.ID NO: 250SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256, SEQ.ID NO: 258SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264, SEQ.ID NO: 266SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272, SEQ.ID NO: 274SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.ID NO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO: 216, SEQ.ID NO:287451850218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224, SEQ.ID NO: 226,SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232, SEQ.ID NO: 234,SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240, SEQ.ID NO: 242,SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248, SEQ.ID NO: 250,SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256, SEQ.ID NO: 258,SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264, SEQ.ID NO: 266,SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272, SEQ.ID NO: 274,SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0055] In at least one embodiment, in an aspect, the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, can comprise: any of the amino acid sequences set forth in: SEQ.ID NO: 2, SEQ.IDNO: 4, SEQ.ID NO: 6, SEQ.ID. NO: 8, SEQ.ID NO: 20, SEQ.ID NO: 22,SEQ.ID NO: 24, SEQ.ID NO: 26, SEQ.ID NO: 28, SEQ.ID NO: 30, SEQ.ID NO: 62, SEQ.ID NO: 64, SEQ.ID NO: 66, SEQ.ID NO: 68, SEQ.ID NO: 70, SEQ.ID NO: 72, SEQ.ID NO: 74, SEQ.ID NO: 76, SEQ.ID NO: 78, SEQ.IDNO: 80, SEQ.ID NO: 82, SEQ.ID NO: 84, SEQ.ID NO: 86, SEQ.ID NO: 88, SEQ.ID NO: 102, SEQ.ID NO: 104, SEQ.ID NO: 106, SEQ.ID NO: 108,SEQ.ID NO: 110, SEQ.ID NO: 112 SEQ.ID NO: 114, SEQ.ID NO: 116,SEQ.ID NO: 187, SEQ.ID NO: 120, SEQ.ID NO: 122, SEQ.ID NO: 124,SEQ.ID NO: 126, or SEQ.ID NO: 128, or an immunologically equivalent portion thereof; and the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, can comprise: any of the amino acid sequences setforth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.ID NO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO:297451850216, SEQ.ID NO: 218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO:224, SEQ.ID NO: 226, SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO:232, SEQ.ID NO: 234, SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO:240, SEQ.ID NO: 242, SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO:248, SEQ.ID NO: 250, SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO:256, SEQ.ID NO: 258, SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO:264, SEQ.ID NO: 266, SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO:272, SEQ.ID NO: 274, SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereof.

[0056] In at least one embodiment, in an aspect, the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, can be a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 25;(b) a nucleic acid sequence having at least 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with the nucleic acid sequence of (a);(c) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to the nucleic acid sequence of (a);(e) a nucleic acid sequence encoding a polypeptide having the amino acid sequences set forth in SEQ.ID NO: 26, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of the amino acid sequence set forth in SEQ.ID NO: 26, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f); and the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, can be a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 9, SEQ.ID NO: 11 , SEQ.ID NO: 13, SEQ.ID NO: 15, SEQ.IDNO: 17, SEQ.ID NO: 31 , SEQ.ID NO: 32, SEQ.ID NO: 34, SEQ.ID NO: 36,307451850SEQ.ID NO: 37, SEQ.ID NO: 39, SEQ.ID NO: 41 , SEQ.ID NO: 42, SEQ.ID NO: 44, SEQ.ID NO: 46, SEQ.ID NO: 47, SEQ.ID NO: 49, SEQ.ID NO: 51 ,SEQ.ID NO: 52, SEQ.ID NO: 54, SEQ.ID NO: 56, SEQ.ID NO: 57, SEQ.IDNO: 59, SEQ.ID NO: 89, SEQ.ID NO: 91 , SEQ.ID NO: 93, SEQ.ID NO: 95,SEQ.ID NO: 97, SEQ.ID NO: 99, SEQ.ID NO: 207, SEQ.ID NO: 209, SEQ.IDNO: 211 , SEQ.ID NO: 213, SEQ.ID NO: 215, SEQ.ID NO: 217, SEQ.ID NO:219, SEQ.ID NO: 221 , SEQ.ID NO: 223, SEQ.ID NO: 225, SEQ.ID NO: 227,SEQ.ID NO: 229, SEQ.ID NO: 231 , SEQ.ID NO: 233, SEQ.ID NO: 235SEQ.ID NO: 237, SEQ.ID NO: 239, SEQ.ID NO: 241 , SEQ.ID NO: 243SEQ.ID NO: 245, SEQ.ID NO: 247, SEQ.ID NO: 249, SEQ.ID NO: 251SEQ.ID NO: 253, SEQ.ID NO: 255, SEQ.ID NO: 257, SEQ.ID NO: 259SEQ.ID NO: 261 , SEQ.ID NO: 263, SEQ.ID NO: 265, SEQ.ID NO: 267SEQ.ID NO: 269, SEQ.ID NO: 271 , SEQ.ID NO: 273, SEQ.ID NO: 343, orSEQ.ID NO: 345;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.ID NO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.IDNO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94,SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.IDNO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO: 216, SEQ.ID NO218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224, SEQ.ID NO: 226SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232, SEQ.ID NO: 234,SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240, SEQ.ID NO: 242,SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248, SEQ.ID NO: 250,SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256, SEQ.ID NO: 258,317451850SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264, SEQ.ID NO: 266, SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272, SEQ.ID NO: 274, SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.ID NO: 35, SEQ.ID NO: 38, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92,SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.IDNO: 208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO:216, SEQ.ID NO: 218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224,SEQ.ID NO: 226, SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232,SEQ.ID NO: 234, SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240,SEQ.ID NO: 242, SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248,SEQ.ID NO: 250, SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256,SEQ.ID NO: 258, SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264,SEQ.ID NO: 266, SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272, SEQ.ID NO: 274, SEQ.ID NO: 344, or SEQ.ID NO: 346or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0057] In at least one embodiment, in an aspect, the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, can be a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 25;(b) a nucleic acid sequence having at least 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with the nucleic acid sequence of (a);(c) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to the nucleic acid sequence of (a);327451850(e) a nucleic acid sequence encoding a polypeptide having the amino acid sequences set forth in SEQ.ID NO: 26, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of the amino acid sequence set forth in SEQ.ID NO: 26, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f); and the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, can be a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 15, or SEQ.ID NO: 17;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 16 or SEQ.ID NO: 18, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 16 or SEQ.ID NO: 18, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0058] In at least one embodiment, in an aspect, the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, can comprise:SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, orSEQ.ID NO: 26, or immunologically equivalent portion thereof; and337451850the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, can comprise: any of the amino acid sequences setforth in SEQ.ID NO: 10, SEQ.IDNO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33,SEQ.ID NO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.IDNO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55,SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.IDNO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO:208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO:216, SEQ.ID NO: 218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO:224, SEQ.ID NO: 226, SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO:232, SEQ.ID NO: 234, SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO:240, SEQ.ID NO: 242, SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO:248, SEQ.ID NO: 250, SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO:256, SEQ.ID NO: 258, SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO:264, SEQ.ID NO: 266, SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO:272, SEQ.ID NO: 274, SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereof.

[0059] In at least one embodiment, in an aspect, the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, can comprise SEQ.ID NO: 26, or an immunologically equivalent portion thereof; and the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, can comprise: any of the amino acid sequences setforth in SEQ.ID NO: 10, SEQ.IDNO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33,SEQ.ID NO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.IDNO: 94, SEQ.ID NO: 9 5, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO: 216, SEQ.ID NO: 218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224, SEQ.ID NO: 226, SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232, SEQ.ID NO: 234, SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO:347451850240, SEQ.ID NO: 242, SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO:248, SEQ.ID NO: 250, SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO:256, SEQ.ID NO: 258, SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO:264, SEQ.ID NO: 266, SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO:272, SEQ.ID NO: 274, SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereof.

[0060] In at least one embodiment, in an aspect, the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, can comprise:SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, orSEQ.ID NO: 26, or an immunologically equivalent portion thereof; and the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, can comprise: any of the amino acid sequences setforth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, or SEQ.ID NO: 100, or an immunogenically equivalent portion thereof.

[0061] In at least one embodiment, in an aspect, at least one of the at least one Ngo1 TbpB protein and / or at least one Ngo2 TbpB protein can be a modulated TbpB protein, such that the Kd between the modulated TbpB protein and native host iron binding protein exceeds the Kd between the native non-modified TbpB protein and the native host iron binding protein by a factor of at least 2x, at least 10x, or at least 100x, wherein, optionally, the native non-modified TbpB protein and the modulated TbpB protein each comprise at least 100 consecutive amino acid residues which are identical to one another, but for one, two, or three modulated amino acid residues comprised within the at least 100 consecutive amino acid residues of the modulated TbpB protein.

[0062] In at least one embodiment, in an aspect, the modulated TbpB protein can be an Ngo1 protein comprising an amino acid sequence selected from SEQ.ID NO: 26, SEQ.ID NO: 62, SEQ.ID NO: 64, SEQ.ID NO: 68, SEQ.ID NO: 70, SEQ.ID NO: 72, SEQ.ID NO: 74, SEQ.ID NO: 76, SEQ.ID NO:80, SEQ.ID NO: 82, SEQ.ID NO: 84, SEQ.ID NO: 86, SEQ.ID NO: 88, SEQ.ID NO: 102, SEQ.ID NO: 108, SEQ.ID NO: 110, SEQ.ID NO: 116, SEQ.ID NO: 122, and SEQ.ID NO: 128, and the Ngo2 protein can be a modulated TbpB protein357451850comprising an amino acid selected from SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO; 48, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO: 216, SEQ.ID NO: 218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224, SEQ.ID NO: 226, SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232, SEQ.ID NO: 234, SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240, SEQ.ID NO: 242, SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248, SEQ.ID NO: 250, SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256, SEQ.ID NO: 258, SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264, SEQ.ID NO: 266, SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272, SEQ.ID NO: 274, SEQ.ID NO: 344, and SEQ.ID NO: 346.

[0063] In at least one embodiment, in an aspect, the modulated TbpB protein can be an Ngo1 protein comprising an amino acid sequence selected from SEQ.ID NO: 26, SEQ.ID NO: 102, and SEQ.ID NO: 108, and the Ngo2 protein can be a modulated TbpB protein comprising an amino acid selected from SEQ.ID NO: 230, SEQ.ID NO: 234, SEQ.ID NO: 238, and SEQ.ID NO: 346.

[0064] In at least one embodiment, in an aspect, the native non-modified protein can be an Ngo1 TbpB protein comprising at least 100 consecutive amino acid residues of SEQ.ID NO: 22.

[0065] In at least one embodiment, in an aspect, the native non-modified TbpB protein can be an Ngo1 TbpB protein comprising at least 100 consecutive amino acid residues of SEQ.ID NO: 22, and the modulated TbpB protein comprises one modulated amino acid residue, the modulated amino acid residue corresponding with aspartic acid amino acid residue 209 (209D) in SEQ.ID NO: 22, and wherein, optionally, aspartic acid amino acid residue 209 (209D) is modulated to replace aspartic acid amino acid residue 209 (209D) in the modulated TbpB protein with a positively charged amino acid residue, optionally, a histidine (H) amino acid residue.

[0066] In at least one embodiment, in an aspect, the native non-modified TbpB protein can be an Ngo1 TbpB protein comprising at least 100 consecutive amino acid residues of SEQ.ID NO: 22, and the modulated TbpB protein comprises SEQ.ID NO: 26.367451850

[0067] In at least one embodiment, in an aspect, the at least two TbpB proteins, or immunogenically equivalent portions thereof, can be recombinantly produced in a host cell, preferably a microbial host cell.

[0068] In at least one embodiment, in an aspect, the at least two TbpB proteins, or immunologically equivalent portions thereof, can be recombinantly produced in a host cell, preferably a microbial host cell, wherein the vaccine formulation is substantially free of host cell constituents, other than the at least two TbpB polypeptides, or immunologically equivalent portions thereof.

[0069] In at least one embodiment, in an aspect, the vaccine formulation can be substantially free of / V. gonorrhoeae constituents, other than the at least two TbpB polypeptides, or immunologically equivalent portions thereof.

[0070] In at least one embodiment, in an aspect, the vaccine formulation can further comprise a pharmaceutically acceptable adjuvant.

[0071] In at least one embodiment, in an aspect, the vaccine formulation can further comprise a pharmaceutically acceptable excipient, carrier, or diluent.

[0072] In another aspect, the present disclosure provides, in accordance with the teachings herein, in at least one embodiment, a method for preparing a vaccine formulation comprising an effective amount of at least two N. gonorrhoeae TbpB proteins, or immunogenically equivalent portions thereof, the vaccine formulation comprising at least one Ngo1 TbpB protein, and at least one Ngo2 TbpB protein, the method comprising:(a) providing a first chimeric nucleic acid sequence comprising as operably linked components:(i) a nucleic acid sequence encoding a first N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, or an mRNA encoding a first N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, wherein the first TbpB protein is an Ngo1 protein; and(ii) one or more nucleic acid sequences capable of controlling expression of the nucleic acid sequence encoding the first TbpB protein, or immunogenically equivalent portion thereof, or the mRNA encoding the first N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, in a host cell;377451850(b) providing a second chimeric nucleic acid sequence comprising as operably linked components:(i) a nucleic acid sequence encoding a second N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, or an mRNA encoding a second N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, wherein the second TbpB protein is an Ngo2 protein; and(ii) one or more nucleic acid sequences capable of controlling expression of the nucleic acid sequence encoding the second TbpB protein, or immunogenically equivalent portion thereof, or the mRNA encoding a first N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, in a host cell;(c) introducing the first and second chimeric nucleic acid sequence each into a separate first and second host cell, or together in a common host cell;(d) growing the first and second separate host cells, or common host cell, to produce the first and second TbpB proteins, or immunogenically equivalent portions thereof, or the mRNA encoding the first or second N. gonorrhoeae TbpB proteins, or an immunogenically equivalent portion thereof;(e) recovering the first and second TbpB proteins, or immunogenically equivalent portions thereof, or the mRNA encoding the first or second N. gonorrhoeae TbpB proteins, or immunogenically equivalent portions thereof; and(f) formulating the first and second TbpB proteins, or immunogenically equivalent portions thereof, or the mRNA encoding the first and second N. gonorrhoeae TbpB proteins, or immunogenically equivalent portions thereof, together with a pharmaceutically acceptable adjuvant to form a vaccine formulation comprising an effective amount of the first and second TbpB protein, or immunogenically equivalent portions thereof, to treat a human susceptible to N. gonorrhoeae infection.

[0073] In another aspect, the present disclosure provides, in accordance with the teachings herein, in at least one embodiment, a method for preparing a vaccine formulation comprising an effective amount of at least two N. gonorrhoeae387451850TbpB proteins, or immunogenically equivalent portions thereof, the vaccine formulation comprising at least one Ngo1 TbpB protein, and at least one Ngo2 TbpB protein, the method comprising:(a) providing a chimeric nucleic acid sequence comprising as operably linked components:(i) a nucleic acid sequence encoding a first N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, or an mRNA encoding a first N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, wherein the first TbpB protein is an Ngo1 TbpB protein;(ii) a nucleic acid sequence encoding a second N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, or an mRNA encoding a second N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, wherein the second TbpB protein is an Ngo2 TbpB protein; and(iii) one or more nucleic acid sequences capable of controlling expression of the nucleic acid sequence encoding the first and the second TbpB protein, or an immunogenically equivalent portion thereof in a host cell;(b) introducing the chimeric nucleic acid sequence into a host cell;(c) growing the host cell to produce the first and second TbpB proteins, or the immunogenically equivalent portions thereof, or the mRNA encoding the first and second N. gonorrhoeae TbpB proteins, or an immunogenically equivalent portion thereof;(d) recovering the first and second TbpB proteins, or the immunogenically equivalent portions thereof, or the mRNA encoding the first or second N. gonorrhoeae TbpB proteins, or immunogenically equivalent portions thereof; and(e) formulating the first and second TbpB proteins, or the immunogenically equivalent portions thereof, or the mRNA encoding the first or second N. gonorrhoeae TbpB proteins, or immunogenically equivalent portions thereof, together with a pharmaceutically acceptable adjuvant to form a vaccine formulation comprising an effective amount of the first and second TbpB protein, or the immunogenically equivalent397451850portions thereof, or the mRNA encoding the first or second / V. gonorrhoeae TbpB proteins, or immunogenically equivalent portions thereof, to treat a human susceptible to N. gonorrhoeae infection.

[0074] In at least one embodiment, in an aspect, the at least one Ngo1 TbpB, or immunologically equivalent portion thereof, can be a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 1 , SEQ.ID NO: 3, SEQ.ID NO: 5, SEQ.ID NO: 7, SEQ.ID NO: 19, SEQ.ID NO: 21 , SEQ.ID NO: 23, SEQ.ID NO: 25, SEQ.ID NO: 27, SEQ.ID NO: 29, SEQ.ID NO: 61 , SEQ.ID NO: 63, SEQ.ID NO: 65, SEQ.ID NO: 67, SEQ.ID NO: 69, SEQ.ID NO: 71 , SEQ.ID NO: 73, SEQ.ID NO: 75, SEQ.ID NO: 77, SEQ.ID NO: 79, SEQ.ID NO: 81 , SEQ.ID NO: 83, SEQ.ID NO: 85, SEQ.ID NO: 87, SEQ.ID NO: 101 , SEQ.ID NO: 103, SEQ.ID NO: 105, SEQ.ID NO: 107, SEQ.ID NO: 109, SEQ.ID NO: 111 , SEQ.ID NO: 113, SEQ.ID NO: 115, SEQ.ID NO: 117, SEQ.ID NO: 119, SEQ.ID NO: 121 , SEQ.ID NO: 123, SEQ.ID NO: 125, or SEQ.ID NO: 127;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 2, SEQ.ID NO: 4, SEQ.ID NO: 6, SEQ.ID. NO: 8, SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, SEQ.ID NO: 26, SEQ.ID NO: 28, SEQ.ID NO: 30, SEQ.ID NO: 62, SEQ.ID NO: 64, SEQ.ID NO: 66, SEQ.ID NO: 68, SEQ.ID NO: 70, SEQ.ID NO: 72, SEQ.ID NO: 74, SEQ.ID NO: 76, SEQ.ID NO: 78, SEQ.ID NO: 80, SEQ.ID NO: 82, SEQ.ID NO: 84, SEQ.ID NO: 86, SEQ.ID NO: 88, SEQ.ID NO: 102, SEQ.ID NO: 104, SEQ.ID NO: 106, SEQ.ID NO: 108, SEQ.ID NO: 110, SEQ.ID NO: 112 SEQ.ID NO: 114, SEQ.ID NO: 116, SEQ.ID NO: 187, SEQ.ID NO: 120, SEQ.ID NO: 122, SEQ.ID NO: 124, SEQ.ID NO: 126, or SEQ.ID NO: 128, or an immunogenically equivalent portion thereof;407451850(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 2, SEQ.ID NO: 4, SEQ.ID NO: 6, SEQ.ID. NO: 8, SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, SEQ.ID NO: 26, SEQ.ID NO: 28, SEQ.ID NO: 30, SEQ.ID NO: 62, SEQ.ID NO: 64, SEQ.ID NO: 66, SEQ.ID NO: 68, SEQ.ID NO: 70, SEQ.ID NO: 72, SEQ.ID NO: 74, SEQ.ID NO: 76, SEQ.ID NO: 78, SEQ.ID NO: 80, SEQ.ID NO: 82, SEQ.ID NO: 84, SEQ.ID NO: 86, SEQ.ID NO: 88, SEQ.ID NO: 102, SEQ.ID NO: 104, SEQ.ID NO: 106, SEQ.ID NO: 108, SEQ.ID NO: 110, SEQ.ID NO: 112 SEQ.ID NO: 114, SEQ.ID NO: 116, SEQ.ID NO: 187, SEQ.ID NO: 120, SEQ.ID NO: 122, SEQ.ID NO: 124, SEQ.ID NO: 126, or SEQ.ID NO: 128, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f); and the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, can be a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 9, SEQ.ID NO: 11 , SEQ.ID NO: 13, SEQ.ID NO: 15, SEQ.ID NO: 17, SEQ.ID NO: 31 , SEQ.ID NO: 32, SEQ.ID NO: 34, SEQ.ID NO: 36, SEQ.ID NO: 37, SEQ.ID NO: 39, SEQ.ID NO: 41 , SEQ.ID NO: 42, SEQ.ID NO: 44, SEQ.ID NO: 46, SEQ.ID NO: 47, SEQ.ID NO: 49, SEQ.ID NO: 51 , SEQ.ID NO: 52, SEQ.ID NO: 54, SEQ.ID NO: 56, SEQ.ID NO: 57, SEQ.ID NO: 59, SEQ.ID NO: 89, SEQ.ID NO: 91 , SEQ.ID NO: 93, SEQ.ID NO: 95, SEQ.ID NO: 97, SEQ.ID NO: 99, SEQ.ID NO: 207, SEQ.ID NO: 209, SEQ.ID NO: 211 , SEQ.ID NO: 213, SEQ.ID NO: 215, SEQ.ID NO: 217, SEQ.ID NO: 219, SEQ.ID NO: 221 , SEQ.ID NO: 223, SEQ.ID NO: 225, SEQ.ID NO: 227,SEQ.ID NO: 229, SEQ.ID NO: 231 , SEQ.ID NO: 233, SEQ.ID NO: 235SEQ.ID NO: 237, SEQ.ID NO: 239, SEQ.ID NO: 241 , SEQ.ID NO: 243SEQ.ID NO: 245, SEQ.ID NO: 247, SEQ.ID NO: 249, SEQ.ID NO: 251SEQ.ID NO: 253, SEQ.ID NO: 255, SEQ.ID NO: 257, SEQ.ID NO: 259SEQ.ID NO: 261 , SEQ.ID NO: 263, SEQ.ID NO: 265, SEQ.ID NO: 267SEQ.ID NO: 269, SEQ.ID NO: 271 , SEQ.ID NO: 273, SEQ.ID NO: 343, or SEQ.ID NO: 345;417451850(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 32, SEQ.IDNO: 34, SEQ.ID NO: 36, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 42,SEQ.ID NO: 44, SEQ.ID NO: 46, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.IDNO: 52, SEQ.ID NO: 54, SEQ.ID NO: 56, SEQ.ID NO: 58, SEQ.ID NO: 60,SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.IDNO: 98, SEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.ID NO: 210, SEQ.ID NO:212, SEQ.ID NO: 214, SEQ.ID NO: 216, SEQ.ID NO: 218, SEQ.ID NO: 220,SEQ.ID NO: 222, SEQ.ID NO: 224, SEQ.ID NO: 226, SEQ.ID NO: 228SEQ.ID NO: 230, SEQ.ID NO: 232, SEQ.ID NO: 234, SEQ.ID NO: 236SEQ.ID NO: 238, SEQ.ID NO: 240, SEQ.ID NO: 242, SEQ.ID NO: 244SEQ.ID NO: 246, SEQ.ID NO: 248, SEQ.ID NO: 250, SEQ.ID NO: 252SEQ.ID NO: 254, SEQ.ID NO: 256, SEQ.ID NO: 258, SEQ.ID NO: 260SEQ.ID NO: 262, SEQ.ID NO: 264, SEQ.ID NO: 266, SEQ.ID NO: 268SEQ.ID NO: 270, SEQ.ID NO: 272, SEQ.ID NO: 274, SEQ.ID NO: 344, orSEQ.ID NO: 346, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12,SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 32, SEQ.IDNO: 34, SEQ.ID NO: 36, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 42, SEQ.ID NO: 44, SEQ.ID NO: 46, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 52, SEQ.ID NO: 54, SEQ.ID NO: 56, SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, orSEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO: 216, SEQ.ID NO: 218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224, SEQ.ID NO: 226, SEQ.ID NO: 228,427451850SEQ.ID NO: 230, SEQ.ID NO: 232, SEQ.ID NO: 234, SEQ.ID NO: 236SEQ.ID NO: 238, SEQ.ID NO: 240, SEQ.ID NO: 242, SEQ.ID NO: 244SEQ.ID NO: 246, SEQ.ID NO: 248, SEQ.ID NO: 250, SEQ.ID NO: 252SEQ.ID NO: 254, SEQ.ID NO: 256, SEQ.ID NO: 258, SEQ.ID NO: 260SEQ.ID NO: 262, SEQ.ID NO: 264, SEQ.ID NO: 266, SEQ.ID NO: 268SEQ.ID NO: 270, SEQ.ID NO: 272, SEQ.ID NO: 274, SEQ.ID NO: 344, orSEQ.ID NO: 346, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0075] In at least one embodiment, in an aspect, the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, can comprise:SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, or SEQ.ID NO:26, or immunologically equivalent portion thereof; and the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, can comprise: any of the amino acid sequences setforth in SEQ.ID NO: 10, SEQ.IDNO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33,SEQ.ID NO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO:208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO:216, SEQ.ID NO: 218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO:224, SEQ.ID NO: 226, SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO:232, SEQ.ID NO: 234, SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO:240, SEQ.ID NO: 242, SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO:248, SEQ.ID NO: 250, SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO:256, SEQ.ID NO: 258, SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO:264, SEQ.ID NO: 266, SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO:272, SEQ.ID NO: 274, SEQ.ID N< : 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereof.437451850

[0076] In at least one embodiment, in an aspect, the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, can comprise:SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, orSEQ.ID NO: 26, or an immunologically equivalent portion thereof; and the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, can comprise: any of the amino acid sequences setforth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, or SEQ.ID NO: 100, or an immunogenically equivalent portion thereof.

[0077] In another aspect, the present disclosure provides, in accordance with the teachings herein, in at least one embodiment, an expression vector comprising a chimeric nucleic acid sequence comprising as operably linked components:(a) a nucleic acid sequence encoding a N. gonorrhoeae TbpB protein, protein, or an immunogenically equivalent portion thereof, or an mRNA encoding a first N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof; and(b) a nucleic acid sequence capable of controlling expression of the nucleic acid sequence encoding the N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, or the mRNA encoding a first N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, in a host cell, wherein the TbpB protein is an Ngo1 TbpB protein, or an Ngo2 TbpB protein.

[0078] In another aspect, the present disclosure provides, in accordance with the teachings herein, in at least one embodiment, an expression vector comprising a chimeric nucleic acid sequence comprising as operably linked components:(a) a nucleic acid sequence encoding a first and second N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, or an mRNA encoding a first and second N. gonorrhoeae TbpB protein, or immunogenically equivalent portions thereof; and447451850(b) one or more nucleic acid sequences capable of separately or jointly controlling expression of the nucleic acid sequence encoding the first and second N. gonorrhoeae TbpB protein, or the immunogenically equivalent portion thereof, or the mRNA encoding the first and second N. gonorrhoeae TbpB protein, or immunogenically equivalent portions thereof, in a host cell, wherein the first and second TbpB protein are an Ngo1 TbpB protein, and an Ngo2 TbpB protein, respectively.

[0079] In at least one embodiment, in an aspect, the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, can be a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 1 , SEQ.ID NO: 3, SEQ.ID NO: 5, SEQ.ID NO: 7, SEQ.ID NO: 19, SEQ.ID NO: 21 , SEQ.ID NO: 23, SEQ.ID NO: 25, SEQ.ID NO: 27, SEQ.ID NO: 29, SEQ.ID NO: 61 , SEQ.ID NO: 63, SEQ.ID NO: 65, SEQ.ID NO: 67, SEQ.ID NO: 69, SEQ.ID NO: 71 , SEQ.ID NO: 73, SEQ.ID NO: 75, SEQ.ID NO: 77, SEQ.ID NO: 79, SEQ.ID NO: 81 , SEQ.ID NO: 83, SEQ.ID NO: 85, SEQ.ID NO: 87, SEQ.ID NO: 101 , SEQ.ID NO: 103, SEQ.ID NO: 105, SEQ.ID NO: 107, SEQ.ID NO: 109, SEQ.ID NO: 111 , SEQ.ID NO: 113, SEQ.ID NO: 115, SEQ.ID NO: 117, SEQ.ID NO: 119, SEQ.ID NO: 121 , SEQ.ID NO: 123, SEQ.ID NO: 125, or SEQ.ID NO: 127;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 2, SEQ.ID NO: 4, SEQ.ID NO: 6, SEQ.ID. NO: 8, SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, SEQ.ID NO: 26, SEQ.ID NO: 28, SEQ.ID NO: 30, SEQ.ID NO: 62, SEQ.ID NO: 64, SEQ.ID NO: 66, SEQ.ID NO: 68, SEQ.ID NO: 70, SEQ.ID NO: 72, SEQ.ID NO: 74, SEQ.ID NO: 76, SEQ.ID NO: 78, SEQ.ID NO: 80, SEQ.ID457451850NO: 82, SEQ.ID NO: 84, SEQ.ID NO: 86, SEQ.ID NO: 88, SEQ.ID NO: 102, SEQ.ID NO: 104, SEQ.ID NO: 106, SEQ.ID NO: 108, SEQ.ID NO: 110, SEQ.ID NO: 112 SEQ.ID NO: 114, SEQ.ID NO: 116, SEQ.ID NO: 187, SEQ.ID NO: 120, SEQ.ID NO: 122, SEQ.ID NO: 124, SEQ.ID NO: 126, or SEQ.ID NO: 128, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 2, SEQ.ID NO: 4, SEQ.ID NO: 6, SEQ.ID. NO: 8, SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, SEQ.ID NO: 26, SEQ.ID NO: 28, SEQ.ID NO: 30, SEQ.ID NO: 62, SEQ.ID NO: 64, SEQ.ID NO: 66, SEQ.ID NO: 68, SEQ.ID NO: 70, SEQ.ID NO: 72, SEQ.ID NO: 74, SEQ.ID NO: 76, SEQ.ID NO: 78, SEQ.ID NO: 80, SEQ.ID NO: 82, SEQ.ID NO: 84, SEQ.ID NO: 86, SEQ.ID NO: 88, SEQ.ID NO: 102, SEQ.ID NO: 104, SEQ.ID NO: 106, SEQ.ID NO: 108, SEQ.ID NO: 110, SEQ.ID NO: 112 SEQ.ID NO: 114, SEQ.ID NO: 116, SEQ.ID NO: 187, SEQ.ID NO: 120, SEQ.ID NO: 122, SEQ.ID NO: 124, SEQ.ID NO: 126, or SEQ.ID NO: 128, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f); and the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, can be a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 9, SEQ.ID NO: 11 , SEQ.ID NO: 13, SEQ.ID NO: 15, SEQ.ID NO: 17, SEQ.ID NO: 31 , SEQ.ID NO: 32, SEQ.ID NO: 34, SEQ.ID NO: 36, SEQ.ID NO: 37, SEQ.ID NO: 39, SEQ.ID NO: 41 , SEQ.ID NO: 42, SEQ.ID NO: 44, SEQ.ID NO: 46, SEQ.ID NO: 47, SEQ.ID NO: 49, SEQ.ID NO: 51 ,SEQ.ID NO: 52, SEQ.ID NO: 54, SEQ.ID NO: 56, SEQ.ID NO: 57, SEQ.IDNO: 59, SEQ.ID NO: 89, SEQ.ID NO: 91 , SEQ.ID NO: 93, SEQ.ID NO: 95,SEQ.ID NO: 97, SEQ.ID NO: 99, SEQ.ID NO: 207, SEQ.ID NO: 209, SEQ.IDNO: 211 , SEQ.ID NO: 213, SEQ.ID NO: 215, SEQ.ID NO: 217, SEQ.ID NO:219, SEQ.ID NO: 221 , SEQ.ID NO: 223, SEQ.ID NO: 225, SEQ.ID NO: 227,SEQ.ID NO: 229, SEQ.ID NO: 231 , SEQ.ID NO: 233, SEQ.ID NO: 235,SEQ.ID NO: 237, SEQ.ID NO: 239, SEQ.ID NO: 241 , SEQ.ID NO: 243,SEQ.ID NO: 245, SEQ.ID NO: 247, SEQ.ID NO: 249, SEQ.ID NO: 251 ,467451850SEQ.ID NO: 253, SEQ.ID NO: 255, SEQ.ID NO: 257, SEQ.ID NO: 259,SEQ.ID NO: 261 , SEQ.ID NO: 263, SEQ.ID NO: 265, SEQ.ID NO: 267,SEQ.ID NO: 269, SEQ.ID NO: 271 , SEQ.ID NO: 273, SEQ.ID NO: 343, or SEQ.ID NO: 345;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.IDNO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45,SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.IDNO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94,SEQ.ID NO: 96, SEQ.ID NO: 98, or SEQ.ID NO: 100, SEQ.ID NO: 208,SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO: 216,SEQ.ID NO: 218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224,SEQ.ID NO: 226, SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232,SEQ.ID NO: 234, SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240,SEQ.ID NO: 242, SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248,SEQ.ID NO: 250, SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256,SEQ.ID NO: 258, SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264,SEQ.ID NO: 266, SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272,SEQ.ID NO: 274, SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.IDNO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94,477451850SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO: 216, SEQ.ID NO:218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224, SEQ.ID NO: 226,SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232, SEQ.ID NO: 234,SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240, SEQ.ID NO: 242,SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248, SEQ.ID NO: 250,SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256, SEQ.ID NO: 258,SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264, SEQ.ID NO: 266,SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272, SEQ.ID NO: 274,SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0080] In another aspect, the present disclosure provides, in accordance with the teachings herein, in at least one embodiment, a host cell comprising a chimeric nucleic acid comprising as operably linked components:(a) a nucleic acid sequence encoding a N. gonorrhoeae TbpB protein, protein, or an immunogenically equivalent portion thereof, or an mRNA encoding a N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof,; and(b) a nucleic acid sequence capable of controlling expression of the nucleic acid sequence encoding the N. gonorrhoeae TbpB protein, or immunogenically equivalent portion thereof, or the mRNA encoding the N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, in a host cell, wherein the TbpB protein is an Ngo1 TbpB protein or an Ngo2 TbpB protein.

[0081] In another aspect, the present disclosure provides, in accordance with the teachings herein, in at least one embodiment, a host cell comprising a chimeric nucleic acid sequence comprising as operably linked components:(a) a nucleic acid sequence encoding a first and second N. gonorrhoeae TbpB protein, or immunogenically equivalent portions thereof, or an mRNA encoding a first and second N. gonorrhoeae TbpB protein, or immunogenically equivalent portions thereof,; and487451850(b) one or more nucleic acid sequences capable of separately or jointly controlling expression of the nucleic acid sequence encoding the first and second N. gonorrhoeae TbpB protein, or the immunogenically equivalent portions thereof in a host cell, or the mRNA encoding the first and second N. gonorrhoeae TbpB protein, or immunogenically equivalent portions thereof, wherein the first and second TbpB protein are selected from an Ngo1 TbpB protein, and an Ngo2 TbpB protein, respectively.

[0082] In another aspect, the present disclosure provides, in accordance with the teachings herein, in at least one embodiment, a mixture comprising a first and second host cell, the first host cell comprising a first chimeric nucleic acid comprising as operably linked components:(a) a nucleic acid sequence encoding a first N. gonorrhoeae TbpB protein, protein, or an immunogenically equivalent portion thereof, or an mRNA encoding a first N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof; and(b) a nucleic acid sequence capable of controlling expression of the nucleic acid sequence encoding the first N. gonorrhoeae TbpB protein, or immunogenically equivalent portion thereof, or the mRNA encoding a first N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, in the first host cell, wherein the first TbpB protein is an Ngo1 TbpB protein; and the second host cell comprising a second chimeric nucleic acid comprising as operably linked components:(c) a nucleic acid sequence encoding a second N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, or an mRNA encoding a second N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof; and(d) a nucleic acid sequence capable of controlling expression of the nucleic acid sequence encoding the second N. gonorrhoeae TbpB protein, or immunogenically equivalent portion thereof, or the mRNA encoding second N. gonorrhoeae TbpB protein, or an497451850immunogenically equivalent portion thereof, in the second host cell, wherein the second TbpB protein is an Ngo2 TbpB protein.

[0083] In at least one embodiment, in an aspect, the Ngo1 TbpB protein, or immunologically equivalent portion thereof, can be a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 1 , SEQ.ID NO: 3, SEQ.ID NO: 5, SEQ.ID NO: 7, SEQ.ID NO: 19, SEQ.ID NO: 21 , SEQ.ID NO: 23, SEQ.ID NO: 25, SEQ.ID NO: 27, SEQ.ID NO: 29, SEQ.ID NO: 61 , SEQ.ID NO: 63, SEQ.ID NO: 65, SEQ.ID NO: 67, SEQ.ID NO: 69, SEQ.ID NO: 71 , SEQ.ID NO: 73, SEQ.ID NO: 75, SEQ.ID NO: 77, SEQ.ID NO: 79, SEQ.ID NO: 81 , SEQ.ID NO: 83, SEQ.ID NO: 85, SEQ.ID NO: 87, SEQ.ID NO: 101 , SEQ.ID NO: 103, SEQ.ID NO: 105, SEQ.ID NO: 107, SEQ.ID NO: 109, SEQ.ID NO: 111 , SEQ.ID NO: 113, SEQ.ID NO: 115, SEQ.ID NO: 117, SEQ.ID NO: 119, SEQ.ID NO: 121 , SEQ.ID NO: 123, SEQ.ID NO: 125, or SEQ.ID NO: 127;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 2, SEQ.ID NO: 4, SEQ.ID NO: 6, SEQ.ID. NO: 8, SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, SEQ.ID NO: 26, SEQ.ID NO: 28, SEQ.ID NO: 30, SEQ.ID NO: 62, SEQ.ID NO: 64, SEQ.ID NO: 66, SEQ.ID NO: 68, SEQ.ID NO: 70, SEQ.ID NO: 72, SEQ.ID NO: 74, SEQ.ID NO: 76, SEQ.ID NO: 78, SEQ.ID NO: 80, SEQ.ID NO: 82, SEQ.ID NO: 84, SEQ.ID NO: 86, SEQ.ID NO: 88, SEQ.ID NO: 102, SEQ.ID NO: 104, SEQ.ID NO: 106, SEQ.ID NO: 108, SEQ.ID NO: 110, SEQ.ID NO: 112 SEQ.ID NO: 114, SEQ.ID NO: 116, SEQ.ID NO: 187, SEQ.ID NO: 120, SEQ.ID NO: 122, SEQ.ID NO: 124, SEQ.ID NO: 126, or SEQ.ID NO: 128, or an immunogenically equivalent portion thereof;507451850(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 2, SEQ.ID NO: 4, SEQ.ID NO: 6, SEQ.ID. NO: 8, SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, SEQ.ID NO: 26, SEQ.ID NO: 28, SEQ.ID NO: 30, SEQ.ID NO: 62, SEQ.ID NO: 64, SEQ.ID NO: 66, SEQ.ID NO: 68, SEQ.ID NO: 70, SEQ.ID NO: 72, SEQ.ID NO: 74, SEQ.ID NO: 76, SEQ.ID NO: 78, SEQ.ID NO: 80, SEQ.ID NO: 82, SEQ.ID NO: 84, SEQ.ID NO: 86, SEQ.ID NO: 88, SEQ.ID NO: 102, SEQ.ID NO: 104, SEQ.ID NO: 106, SEQ.ID NO: 108, SEQ.ID NO: 110, SEQ.ID NO: 112 SEQ.ID NO: 114, SEQ.ID NO: 116, SEQ.ID NO: 187, SEQ.ID NO: 120, SEQ.ID NO: 122, SEQ.ID NO: 124, SEQ.ID NO: 126, or SEQ.ID NO: 128, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f); and the Ngo2 TbpB protein, or immunologically equivalent portion thereof, can be a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 9, SEQ.ID NO: 11 , SEQ.ID NO: 13, SEQ.ID NO: 15, SEQ.ID NO: 17, SEQ.ID NO: 31 , SEQ.ID NO: 32, SEQ.ID NO: 34, SEQ.ID NO: 36, SEQ.ID NO: 37, SEQ.ID NO: 39, SEQ.ID NO: 41 , SEQ.ID NO: 42, SEQ.ID NO: 44, SEQ.ID NO: 46, SEQ.ID NO: 47, SEQ.ID NO: 49, SEQ.ID NO: 51 , SEQ.ID NO: 52, SEQ.ID NO: 54, SEQ.ID NO: 56, SEQ.ID NO: 57, SEQ.ID NO: 59, SEQ.ID NO: 89, SEQ.ID NO: 91 , SEQ.ID NO: 93, SEQ.ID NO: 95, SEQ.ID NO: 97, SEQ.ID NO: 99, SEQ.ID NO: 207, SEQ.ID NO: 209, SEQ.ID NO: 211 , SEQ.ID NO: 213, SEQ.ID NO: 215, SEQ.ID NO: 217, SEQ.ID NO: 219, SEQ.ID NO: 221 , SEQ.ID NO: 223, SEQ.ID NO: 225, SEQ.ID NO: 227,SEQ.ID NO: 229, SEQ.ID NO: 231 , SEQ.ID NO: 233, SEQ.ID NO: 235SEQ.ID NO: 237, SEQ.ID NO: 239, SEQ.ID NO: 241 , SEQ.ID NO: 243SEQ.ID NO: 245, SEQ.ID NO: 247, SEQ.ID NO: 249, SEQ.ID NO: 251SEQ.ID NO: 253, SEQ.ID NO: 255, SEQ.ID NO: 257, SEQ.ID NO: 259SEQ.ID NO: 261 , SEQ.ID NO: 263, SEQ.ID NO: 265, SEQ.ID NO: 267SEQ.ID NO: 269, SEQ.ID NO: 271 , SEQ.ID NO: 273 SEQ.ID NO: 343, orSEQ.ID NO: 345;517451850(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.IDNO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45,SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.IDNO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94,SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.IDNO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO: 216, SEQ.ID NO:218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224, SEQ.ID NO: 226,SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232, SEQ.ID NO: 234SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240, SEQ.ID NO: 242SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248, SEQ.ID NO: 250SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256, SEQ.ID NO: 258SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264, SEQ.ID NO: 266SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272, SEQ.ID NO: 274SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.IDNO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO: 216, SEQ.ID NO: 218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224, SEQ.ID NO: 226, SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232, SEQ.ID NO: 234,527451850SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240, SEQ.ID NO: 242,SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248, SEQ.ID NO: 250,SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256, SEQ.ID NO: 258,SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264, SEQ.ID NO: 266,SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272, SEQ.ID NO: 274,SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0084] In another aspect, the present disclosure provides, in accordance with the teachings herein, in at least one embodiment, a use of an Ngo1 TbpB protein, or an immunogenically equivalent portion thereof, together with an Ngo2 TbpB protein, or an immunogenically equivalent portion thereof, or a use of or an mRNA encoding an Ngo1 TbpB protein, or an immunogenically equivalent portion thereof, together with an mRNA encoding an Ngo2 TbpB protein, or an immunogenically equivalent portion thereof, to prepare a vaccine formulation comprising the Ngo1 and Ngo2 TbpB proteins, or immunogenically equivalent portions thereof, or a vaccine formulation comprising the mRNAs encoding the Ngo1 and Ngo2 TbpB polypeptides, or immunologically equivalent portions thereof, together with a pharmaceutically acceptable adjuvant.

[0085] In another aspect, the present disclosure provides, in accordance with the teachings herein, in at least one embodiment, an Ngo1 TbpB protein, or an immunogenically equivalent portion thereof, together with an Ngo2 TbpB protein, or an immunogenically equivalent portion thereof, or an mRNA encoding an Ngo1 TbpB protein, or an immunogenically equivalent portion thereof, together with an mRNA encoding an Ngo2 TbpB protein, or an immunogenically equivalent portion thereof, for use in the preparation of a vaccine formulation comprising the Ngo1 and Ngo2 TbpB proteins, or immunogenically equivalent portions thereof, or a vaccine formulation comprising the mRNAs encoding the Ngo1 and Ngo2 TbpB polypeptides, or immunologically equivalent portions thereof, together with a pharmaceutically acceptable adjuvant.

[0086] Other features and advantages will become apparent from the following detailed description. It should be understood, however, that the detailed description, while indicating preferred implementations of the disclosure, are given537451850by way of illustration only, since various changes and modifications within the spirit and scope of the disclosure will become apparent to those of skill in the art from the detailed description.BRIEF DESCRIPTION OF THE DRAWINGS

[0087] The disclosure is in the hereinafter provided paragraphs described, by way of example, in relation to the attached figures. The figures provided herein are provided for a better understanding of the example embodiments and to show more clearly how the various embodiments may be carried into effect. The figures are not intended to limit the present disclosure.

[0088] FIGS. 1A and 1B are two phylogenetic trees of multiple unique / V. gonorrhoeae and N. meningitidis TbpB protein sequences collected from publicly available N. gonorrhoeae and N. meningitidis TbpB sequences available in online databases, combined with certain TbpB sequences obtained from a local collection. Indicated in the phylogenetic tree in FIG. 1A are separate N. gonorrhoeae phylogenetic clusters denoted as: “Ngo 1”, “Ngo 2”, and N. meningitidis phylogenetic clusters denoted as “Nme 1”, “Nme 2”, “Nme 3”, “Nme 4”, and “Isotype I”. Further indicated are the phylogenetic location of multiple N. gonorrhoeae TbpB polypeptides denoted as: “48627” (see: SEQ.ID NO: 20), “WHO Y” (see: SEQ.ID NO: 28), “27464” (see: SEQ.ID NO: 6), and “27230” (see: SEQ.ID NO: 2), and “41385” (see: SEQ.ID NO: 10). Indicated in the phylogenetic tree in FIG. 1B are separate N. gonorrhoeae phylogenetic clusters denoted as: “Ngo 1” and “Ngo 2”. Further indicated are the phylogenetic location of multiple N. gonorrhoeae TbpB polypeptides denoted as: “48627” (see: SEQ.ID NO: 20), “27464” (see: SEQ.ID NO: 6), “27230” (see: SEQ.ID NO: 2), “41385” (see: SEQ.ID NO: 10), “32079” (see: SEQ.ID NO: 38), “757977” (see: SEQ.ID NO: 53), “32017” (see: SEQ.ID NO: 33), “36247” (see: SEQ.ID NO: 43), “47626” (see: SEQ.ID NO: 48), and “78648” (see: SEQ.ID NO: 58). Branches are in units of average substitutions per site, with the magnitudes indicated by the scale bar in the bottom right corners of FIGS. 1A and 1B.

[0089] FIGS. 2A, 2B, and 2C depict schematics of workflows relating to the recombinant expression of TbpB proteins in Escherichia coli using a cytoplasmic expression system (FIG. 2A), and a secretion system (FIG. 2B), as well as certain experimental results, notably, a solid-phase filter obtained in the performance of a547451850solid-phase binding assay to detect binding of TbpB proteins to human transferrin (hTf), notably binding to wildtype TbpB protein and certain mutant TbpB proteins of various i\i. gonorrhoeae strains, notably (i) for Ngo1 strain 48627 wildtype (WT) and the mutant TbpB proteins: (K186A), (K186E), (K196A), (R200A), (R200D), (D209A), and (D209H) (non-binding mutants: (R200A); (R200D); (D209H)); (ii) for Ngo2 strain 41385, wildtype (WT) and the mutant TbpB proteins: (K199A), (K199E), (K208A), (R209A), (G222A), (G222R), (E223A), and (E223R) (nonbinding mutants: (G222R); E223R); (iii) for Ngo1 strain 27230, wildtype (WT) and the mutant TbpB proteins: (K182A), (K182E), (K192A), (K196A), (R196D), (D205A), (D205H), and (D205K) (non-binding mutants: (R196D); (D205A); (D205K); (iv) for Ngo1 strain 27464, wildtype (WT) and the mutant TbpB proteins: (R185A), (R185D), (K195A), (K196A), (K195E), (D208A), and (D208R) (nonbinding mutants: (R208A); D208R) ; and (v) for Ngo2 strain WHO Y wildtype (WT) and the mutant TbpB proteins: (K188A), (K188E), (K192A), (K198A), (R202A), (R202D), (D225A), (D225H), and (D225K) (non-binding mutants: (K188E); (R202D); (D225K), (FIG. 2C).

[0090] FIGS. 3A, 3B, 3C, 3D, 3E, 3F, 3G, 3H, 3I, and 3J depict various graphs representing certain experimental results, notably graphs obtained in the performance of experimental assays to evaluate the in vivo efficacy of vaccine formulations comprising an Ngo1 TbpB protein and an Ngo2 TbpB protein, notably a colonization assay following immunization and a challenge with a WHO L strain (FIGS. 3A, 3B) or a WHO F strain (FIGS. 3F, 3G), as well as a colony forming units (CFU) enumeration assay following immunization and a challenge with a WHO L strain in control animals (FIG. 3C) and vaccinated animals (FIG. 3D), or a WHO F strain in control animals (FIG. 3H) and vaccinated animals (FIG. 3I), and a total bacterial burden enumeration assay following immunization and a challenge with a WHO L strain (FIG. 3E), or a challenge with a WHO F strain (FIG. 3J).

[0091] FIGS. 4A, 4B, 4C, and 4D depict various graphs representing certain experimental results, notably graphs obtained in the performance of experimental assays to evaluate the in vivo efficacy of vaccine formulations comprising an Ngo1 TbpB protein and an Ngo2 TbpB protein, notably serum immunogenicity assays evaluating immune mouse serum detecting Ngo2 TbpB antibodies (denoted as Anti-41385) (FIG. 4A), Ngo1 TbpB antibodies (denoted as Anti-48627) (FIG. 4B), and cross reactivity against a panel of variant TbpBs from N. gonorrhoeae strains557451850(41385, 48627 MS11 , FA1090, WHO L, WHO M, WHO O, WHO, P, WHO V, WHO X, WHO Y, FA19, WHO F, WHO G, and WHO W) and N. meningitidis strains (P3006, M978, N206, M922, MC58, M990, H4476, N224, S3131 , B16B6, and MBP) (FIG. 4C), showing comparable human transferrin binding to all coated TbpB proteins (FIG. 4D).

[0092] FIGS. 5A, 5B and 5C depict various graphs representing certain experimental results, notably graphs obtained in the performance of experimental assays to evaluate the in vivo efficacy of vaccine formulations comprising a wildtype Ngo1 TbpB protein (Ngo_1 ) and a modulated Ngo1 TbpB protein (D209H), notably an immunogenicity assay detecting TbpB antibodies from serum from immunized transgenic mice expressing human transferrin (FIG. 5A), an hTf blocking assay using serum from wildtype (WT) mice and transgenic mice expressing human transferrin immunized with the wildtype and modulated TbpB proteins (FIG. 5B) and a genital colonization assay where transgenic mice expressing human transferrin were immunized with the different vaccine formulations and challenged with N. gonorrhoeae WHO L (FIG. 5C).

[0093] FIGS. 6A, 6B, and 6C depict various graphs representing certain experimental results, notably graphs obtained in the performance of experimental assays to evaluate the in vivo efficacy of vaccine formulations comprising a modulated Ngo1 TbpB protein (Ngo 48627 D209H TbpB (SEQ.ID NO: 26), denoted a “D”) and one of two modulated Ngo2 TbpB proteins (Ngo 41385 G222R TbpB (SEQ.ID NO: 16), denoted as “G” or Ngo 41385 E223R TbpB (SEQ.ID NO: 18), denoted as “E”), notably, a colonization assay in wild type mice where immunized mice where challenged with N. gonorrhoeae strain WHO L (FIGS. 6A, 6B) and the subsequent CFU enumeration of recovered bacteria (FIG. 6C).

[0094] FIGS. 7A, 7B, and 7C depict various heat maps representing certain experimental results, notably heatmaps obtained in the performance of experimental assays, notably modified enzyme-linked immunosorbent assays (ELISA) to evaluate the in vivo efficacy of vaccine formulations comprising various Ngo1 TbpB proteins (“27464”, “48627”, “WHO Y”, and “27236”) and Ngo2 TbpB proteins (“41385”) and combinations thereof, notably, a heat map evaluating cross-reactivity of serum of rabbits immunized with various vaccine formulations (identified below the heat map in horizontal direction) against various N. gonorrhoeae and N. meningitidis TbpBs (identified to the left of the heat map in567451850vertical direction) (FIG. 7A), a heat map evaluating blocking ability of serum from rabbits immunized with various vaccine formulations (identified in horizontal direction) against various N. gonorrhoeae and N. meningitidis TbpBs (identified in vertical direction) using human transferrin conjugated to horse radish peroxidase (FIG. 7C), and a control to establish comparable amounts of the different TbpBs were present in the ELISA assays (FIG. 7B).

[0095] FIGS. 8A, 8B, 8C, 8D, 8E, 8F, 8G, and 8H depict various graphs representing certain experimental results, notably graphs obtained in the performance of experimental assays, notably serum bactericidal assays (SBAs) to evaluate the efficacy of vaccine formulations comprising various Ngo1 TbpB proteins (“27464”, “48627”, “WHO Y”, and “27236”) and Ngo2 TbpB proteins (“41385”) and combinations thereof, notably, graphs evaluating cross-bactericidal activity of serum of rabbits immunized with various vaccine formulations (identified in horizontal direction) against various N. gonorrhoeae strains, with the gonococcal porin type and TbpB cluster and the percent normal human serum used for bactericidal killing noted (identified above each graph). Strains tested include MS11 strain (FIG. 8A), FA1090 strain (FIG. 8B), FA19 strain (FIG. 8C), WHO G strain (FIG. 8D), WHO P strain (FIG. 8E), WHO W strain (FIG. 8F), WHO X strain (FIG. 8G), and WHO Y strain (FIG. 8H).

[0096] The figures together with the following detailed description make apparent to those skilled in the art how the disclosure may be implemented in practice.DETAILED DESCRIPTION

[0097] The following paragraphs are intended to introduce the reader to the more detailed description, not to define or limit the claimed subject matter of the present disclosure.

[0098] Various compositions, methods, or processes will be described below to provide an example of an embodiment of each claimed subject matter. No embodiment described below limits any claimed subject matter and any claimed subject matter may cover processes, compositions or methods that differ from those described below. The claimed subject matter is not limited to compositions, processes or methods having all of the features of any one composition, method or process described below or to features common to577451850multiple or all of the compositions, methods or processes described below. It is possible that a composition, method, or process described below is not an embodiment of any claimed subject matter. Any subject matter disclosed in a composition, method, or process described below that is not claimed in this document may be the subject matter of another protective instrument, for example, a continuing patent application, and the applicant(s), inventor(s) orowner(s) do not intend to abandon, disclaim or dedicate to the public any such subject matter by its disclosure in this document.

[0099] As used herein and in the claims, the singular forms, such “a”, “an” and “the” include the plural reference and vice versa unless the context clearly indicates otherwise. Throughout this specification, unless otherwise indicated, “comprise,” “comprises” and “comprising” are used inclusively rather than exclusively, so that a stated integer or group of integers may include one or more other non-stated integers or groups of integers. The term “or” is inclusive unless modified, for example, by “either”.

[0100] When ranges are used herein for physical properties, such as molecular weight, or chemical properties, such as chemical formulae, all combinations and sub-combinations of ranges and specific embodiments therein are intended to be included. Other than in the operating examples, or where otherwise indicated, all numbers expressing quantities of ingredients or reaction conditions used herein should be understood as modified in all instances by the term “about.” The term “about” when referring to a number or a numerical range means that the number or numerical range referred to is an approximation within experimental variability (or within statistical experimental error), and thus the number or numerical range may vary between 1 % and 15% of the stated number or numerical range, as will be readily recognized by context. Furthermore, any range of values described herein is intended to specifically include the limiting values of the range, and any intermediate value or sub-range within the given range, and all such intermediate values and sub-ranges are individually and specifically disclosed (e.g., a range of 1 to 5 includes 1 , 1.5, 2, 2.75, 3, 3.90, 4, and 5). Similarly, otherterms of degree such as "substantially" and "approximately" as used herein mean a reasonable amount of deviation of the modified term such that the end result is not significantly changed. These terms of degree should be587451850construed as including a deviation of the modified term if this deviation would not negate the meaning of the term it modifies.

[0101] Unless otherwise defined, scientific and technical terms used in connection with the formulations described herein shall have the meanings that are commonly understood by those of ordinary skill in the art. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the present invention, which is defined solely by the claims.

[0102] All publications, patents and patent applications are herein incorporated by reference in their entirety to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference in its entirety.Terms and definitions

[0103] The terms “nucleic acid”, or “nucleic acid sequence”, as used herein, refer to a sequence of nucleoside or nucleotide monomers, consisting of naturally occurring bases, sugars and intersugar (backbone) linkages. The term also includes modified or substituted sequences comprising non-naturally occurring monomers or portions thereof. The nucleic acids of the present disclosure may be deoxyribonucleic nucleic acids (DNA) or ribonucleic acids (RNA) and may include naturally occurring bases including adenine, guanine, cytosine, thymidine, and uracil. The nucleic acids may also contain modified bases. Examples of such modified bases include aza and deaza adenine, guanine, cytosine, thymidine and uracil, and xanthine and hypoxanthine. A sequence of nucleotide or nucleoside monomers may be referred to as a polynucleotide sequence, nucleic acid sequence, a nucleotide sequence, or a nucleoside sequence.

[0104] The terms “polypeptide” and “protein”, as may be used interchangeably herein, in conjunction with a reference SEQ.ID NO, refer to any and all polypeptides and proteins comprising a sequence of amino acid residues which is (i) substantially identical to the amino acid sequence constituting the polypeptide having such reference SEQ.ID NO, or (ii) encoded by a nucleic acid sequence capable of hybridizing under at least moderately stringent conditions to any nucleic acid sequence encoding the polypeptide having such reference597451850SEQ.ID NO, but for the use of synonymous codons. A sequence of amino acid residues may be referred to as an amino acid sequence, or polypeptide sequence.

[0105] The terms “nucleic acid sequence encoding a polypeptide” and “nucleic acid sequence encoding a protein”, as used herein in conjunction with a reference SEQ.ID NO, refer to any and all nucleic acid sequences encoding a polypeptide or protein having such reference SEQ.ID NO. Nucleic acid sequences encoding a polypeptide, in conjunction with a reference SEQ.ID NO, further include any and all nucleic acid sequences which (i) encode polypeptides that are substantially identical to the polypeptide having such reference SEQ.ID NO; or (ii) hybridize to any nucleic acid sequences encoding polypeptides having such reference SEQ.ID NO under at least moderately stringent hybridization conditions or which would hybridize thereto under at least moderately stringent conditions but for the use of synonymous codons.

[0106] The terms “nucleic acid sequence encoding TbpB”, and “nucleic acid sequence encoding a “TbpB polypeptide”, “nucleic acid sequence encoding a TbpB protein” as may be used interchangeably herein, refer to any and all nucleic acid sequences encoding a TbpB polypeptide, including, for example, SEQ.ID NO: 1. Nucleic acid sequences encoding a TbpB polypeptide further include any and all nucleic acid sequences which (i) encode polypeptides that are substantially identical to the TbpB polypeptide sequences set forth herein; or (ii) hybridize to any TbpB nucleic acid sequences set forth herein under at least moderately stringent hybridization conditions or which would hybridize thereto under at least moderately stringent conditions but for the use of synonymous codons.

[0107] The terms “TbpB protein” or “TbpB polypeptide”, as may be used herein, interchangeably refer to any and all protein comprising a sequence of amino acid residues which is (i) substantially identical to the amino acid sequences constituting any TbpB polypeptide set forth herein, including, for example, SEQ.ID NO: 2, or (ii) encoded by a nucleic acid sequence capable of hybridizing under at least moderately stringent conditions to any nucleic acid sequence encoding any TbpB protein set forth herein, but for the use of synonymous codons.

[0108] By the term “substantially identical” it is meant that two amino acid sequences preferably are at least 70% identical, or preferably at least 80% identical, and more preferably are at least 85% or 90% identical, for example, 90%, 91 %, 92%, 93%, or 94% identical, and most preferably at least 95% identical, for607451850example, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, or 99.9% identical. In order to determine the percentage of identity between two amino acid sequences the amino acid sequences of such two sequences are aligned, using for example the alignment method of Needleman and Wunsch (J. Mol. Biol., 1970, 48: 443), as revised by Smith and Waterman (Adv. Appl. Math., 1981 , 2: 482) so that the highest order match is obtained between the two sequences and the number of identical amino acids is determined between the two sequences. Methods to calculate the percentage identity between two amino acid sequences are generally art recognized and include, for example, those described by Carillo and Lipton (SIAM J. Applied Math., 1988, 48:1073) and those described in Computational Molecular Biology, Lesk, e.d. Oxford University Press, New York, 1988, Biocomputing: Informatics and Genomics Projects. Generally, computer programs will be employed for such calculations. Computer programs that may be used in this regard include, but are not limited to, GCG (Devereux et al., Nucleic Acids Res., 1984, 12: 387) BLASTP, BLASTN and FASTA (Altschul et a!., J. Mol. Biol., 1990:215:403). A particularly preferred method for determining the percentage identity between two polypeptides involves the Clustal W algorithm (Thompson, J. D., Higgines, D. G. and Gibson T. J / , 1994, Nucleic Acid Res 22(22): 4673-4680 together with the BLOSUM 62 scoring matrix (Henikoff S & Henikoff, J G, 1992, Proc. Natl. Acad. Sci. USA 89: 10915-10919 using a gap opening penalty of 10 and a gap extension penalty of 0.1 , so that the highest order match obtained between two sequences wherein at least 50% of the total length of the two sequences is involved in the alignment.

[0109] By “at least moderately stringent hybridization conditions” it is meant that conditions are selected which promote selective hybridization between two complementary nucleic acid molecules in solution. Hybridization may occur to all or a portion of a nucleic acid sequence molecule. The hybridizing portion is typically at least 15 (e.g., 20, 25, 30, 40 or 50) nucleotides in length. Those skilled in the art will recognize that the stability of a nucleic acid duplex, or hybrids, is determined by the Tm, which in sodium containing buffers is a function of the sodium ion concentration and temperature (Tm=81.5° C.-16.6 (Log10 [Na+])+0.41 (% (G+C)- 600 / I), or similar equation). Accordingly, the parameters in the wash conditions that determine hybrid stability are sodium ion concentration and temperature. In order to identify molecules that are similar, but not identical,617451850to a known nucleic acid molecule a 1 % mismatch may be assumed to result in about a 1 ° C. decrease in Tm, for example if nucleic acid molecules are sought that have a >95% identity, the final wash temperature will be reduced by about 5° C. Based on these considerations those skilled in the art will be able to readily select appropriate hybridization conditions. In preferred embodiments, stringent hybridization conditions are selected. By way of example the following conditions may be employed to achieve stringent hybridization: hybridization at 5x sodium chloride / sodium citrate (SSC) / 5xDenhardt's solution / 1.0% SDS at Tm (based on the above equation) -5° C, followed by a wash of 0.2xSSC / 0.1 % SDS at 60° C. Moderately stringent hybridization conditions include a washing step in 3xSSC at 42° C. It is understood however that equivalent stringencies may be achieved using alternative buffers, salts, and temperatures. Additional guidance regarding hybridization conditions may be found in: Current Protocols in Molecular Biology, John Wiley & Sons, N.Y., 1989, 6.3.1.-6.3.6 and in: Sambrook et al., Molecular Cloning, a Laboratory Manual, Cold Spring Harbor Laboratory Press, 1989, Vol. 3.

[0110] The term “functional variant”, as used herein in reference to polynucleotides or polypeptides, refers to polynucleotides or polypeptides capable of performing the same function as a noted reference polynucleotide or polypeptide. Thus, for example, a functional variant of the polypeptide set forth in SEQ.ID NO: 2, refers to a polypeptide capable of performing the same function as the polypeptide set forth in SEQ.ID NO: 2. Functional variants include modified a polypeptide wherein, relative to a noted reference polypeptide, the modification includes a substitution, deletion, or addition of one or more amino acids. In some embodiments, substitutions are those that result in a replacement of one amino acid with an amino acid having similar characteristics. Such substitutions include, without limitation (i) glutamic acid and aspartic acid; (i) alanine, serine, and threonine; (iii) isoleucine, leucine, and valine, (iv) asparagine and glutamine, and (v) tryptophan, tyrosine, and phenylalanine.

[0111] The term “chimeric”, as used herein in the context of nucleic acids, refers to at least two linked nucleic acids which are not naturally linked. Chimeric nucleic acids include linked nucleic acids of different natural origins. For example, a nucleic acid constituting a microbial promoter linked to a nucleic acid encoding a plant polypeptide is considered chimeric. Chimeric nucleic acids also may comprise nucleic acids of the same natural origin, provided they are not naturally627451850linked. For example, a nucleic acid constituting a promoter obtained from a particular cell-type may be linked to a nucleic acid encoding a polypeptide obtained from that same cell-type, but not normally linked to the nucleic acid constituting the promoter. Chimeric nucleic acids also include nucleic acids comprising any naturally occurring nucleic acids linked to any non-naturally occurring nucleic acids.

[0112] The term “phylogenetic cluster”, as used herein, refers to a group of evolutionary related polypeptide sequences. In order to determine whether two polypeptides belong to the same phylogenetic cluster, an evolutionary tree containing multiple branches (for example, at least 5, 7, 10, 15, or 20 branches), can be constructed using multiple more or less similar input polypeptide sequences (for example, preferably at least 25, at least 50, at least 100, or at least 1 ,000, polypeptide sequences). Upon inspection of the phylogenetic tree, the evolutionary relationship of the polypeptide sequences can be evaluated. Polypeptides belonging to the same phylogenetic cluster are polypeptides located on a particular branch descended from a common ancestor on a phylogenetic tree. Those skilled in the art will be familiar with software programs to assist in the automated generation of phylogenetic trees based on polypeptide sequence input. Suitable phylogenetic tree construction software includes, for example, sequence alignment software such as MAFFT (e.g., v7.450 orv4.475) (Katoh, K. etal., 2002, Nucleic Acids Research, 30 (14), 3059-3066) which can, for example, be using the E-INS-i algorithm; evolutionary modeling software to identify an appropriate evolutionary model such as ProtTest (e.g., v3.4.2) (Darriba D, Taboada GL, Doallo R, Posada D. ProtTest 3: fast selection of best-fit models of protein evolution. Bioinformatics, 27:1164-1165, 2011); and phylogenetic tree reconstruction software such as, for example, PhyML (e.g., v3.3.20190909) (Guindon S., et al., 2010, Systematic Biology, 59(3):307-21 , 2010) and RAxML (e.g., v8.2.12) (Stamakis, A. Bioinformatics, Volume 30, Issue 9, May 2014, Pages 1312— 1313, doi.org / 10.1093 / bioinformatics / btu033), the LG model of evolution (https: / / pubmed.ncbi.nlm.nih.gov / 18367465), and Navargator to perform k- medoids clustering (https: / / www.compsysbio.org / navargator). FIGS. 1A and 1 B show example phylogenetic trees of TbpB polypeptide sequences. Indicated in the example phylogenetic tree of FIG. 1A are example phylogenetic clusters Ngo1 , Ngo2, Nme1 , Nme2, Nme3, Nme4 and Isotype I, and indicated in FIG. 1B are637451850example phylogenetic clusters Ngo1 and Ngo2. The example phylogenetic trees of FIGS. 1A and 1B can be constructed as described further in Example 1 hereto. To evaluate the evolutionary relationship of a TbpB polypeptide whose evolutionary relationship to other TbpB polypeptides is unknown, the TbpB polypeptide sequence thereof may be included in the input TbpB polypeptide sequences used to construct a phylogenetic tree, for example, the input TbpB sequences used to construct example phylogenetic trees of FIGS. 1 A and 1 B, and a phylogenetic tree including the TbpB polypeptide can be constructed. The constructed phylogenetic tree can then be evaluated with respect to the location of the TbpB polypeptide, including its location within a phylogenetic cluster.

[0113] “Phylogenetic cluster Ngo1”, as used herein, refers to a phylogenetic cluster designated as “Ngo1” in the example phylogenetic trees of FIGS. 1A and 1B. The phylogenetic trees may be constructed using a plurality of TbpB polypeptide sequences, as described in Example 1 hereto. Example TbpB polypeptide sequences in phylogenetic cluster Ngo1 include, for example, TbpB polypeptide sequences having: SEQ.ID NO: 20, SEQ.ID NO: 28, SEQ.ID NO: 6), and “27230” (see: SEQ.ID NO: 2), as set forth herein.

[0114] “Phylogenetic cluster Ngo2”, as used herein, refers to a phylogenetic cluster designated as “Ngo2” in the phylogenetic trees of FIGS. 1A and 1B. The phylogenetic trees may be constructed using a plurality of TbpB polypeptide sequences, as described in Example 1 hereto. Example TbpB polypeptide sequences in phylogenetic cluster Ngo2 include, for example, TbpB polypeptide sequences having SEQ.ID NO: 10, SEQ.ID NO: 33, SEQ.ID NO: 38, SEQ.ID NO: 43, SEQ.ID NO: 48, SEQ.ID NO: 53, and SEQ.ID NO: 58, as set forth herein.

[0115] The term “Ngo1 TbpB protein”, as used herein, refers to a TbpB protein located on a branch of a phylogenetic tree of TbpB proteins within phylogenetic cluster Ngo1 .

[0116] The term “Ngo2 TbpB protein”, as used herein, refers to a TbpB protein located on a branch of a phylogenetic tree of TbpB proteins within phylogenetic cluster Ngo2.

[0117] The terms “Neisseria gonorrhoeae” or “N. gonorrhoeae”, as used herein, refer to any bacteria belonging the bacterial species taxonomically classified as such and include any subspecies, strains, variants, serogroups, serotypes, or genotypes.647451850

[0118] The term “effective amount”, as used herein, refers to an amount of an active agent or pharmaceutical formulation, including a vaccine formulation, sufficient to induce a desired biological or therapeutic effect, including a prophylactic effect. Such effect can include an effect with respect to the signs, symptoms or causes of a disorder, or disease or any other desired alteration of a biological system. The effective amount can vary depending, for example, on the health condition, injury stage, disorder stage, or disease stage, of the human or test animal, being treated, timing of the administration, manner of the administration, age of the human or test animal, weight of the human or test animal, and the like, all of which can be determined by those of skill in the art.

[0119] The term “immunologically equivalent”, as used herein, refers to a molecule that is capable of eliciting a humoral immune response in the form of the production of native polyclonal antibodies in a subject human or test animal when administered thereto, wherein the binding specificity to the native polyclonal antibodies is comparable to the specificity of native polyclonal antibodies produced when a reference molecule is administered to a human or subject test animal. For example, immunologically equivalent portions of a reference full length TbpB polypeptide include immunogenic portions of a TbpB polypeptide which when administered to a human or subject test animal elicit a humoral immune response in the form of the production of native antibodies with a specificity to a TbpB polypeptide which is comparable to the binding specificity for a TbpB polypeptide of native antibodies obtained when the reference full length TbpB is administered to the human or subject test animal. Immunologically equivalent portions of full length TbpB polypeptides can vary in length and may, for example, include polypeptides comprising or consisting of at least 10, at least or up to 15, at least or up to 20, at least or up to 30, at least or up to 50, or at least or up to 60 consecutive amino acid residues which are identical to a portion of a TbpB polypeptide. Furthermore, immunologically equivalent portions of full length TbpB polypeptides include polypeptides which are at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9% identical to a full length TbpB polypeptide. To compare binding specificity between a reference molecule and an immunologically equivalent molecule a radioimmune assay (RIA) may be used, and the extent of binding may be measured. The dissociation constant of an657451850immunologically equivalent molecule is preferably at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of the dissociation constant of a reference molecule.

[0120] The term “cross-protective”, as used herein, refers to a vaccine formulation capable of providing protection against infection by multiple strains (e.g., 2, 3, 4, 5, or more strains) of a pathogenic microbial organism, for example, strains belonging to multiple serogroups or serotypes. A cross-protective vaccine formulation may comprise multiple antigenic substances, or a single antigenic substance, for example, multiple immunogenic polypeptides or a single immunogenic polypeptide. To evaluate cross-protection, a vaccine formulation comprising one or more immunogenic substances obtained from a selected strain of a microbial organism, a TbpB protein, for example, may be used to immunize a subject test animal or human. The subject test animal or human then may be exposed to (challenged with) another pathogenic strain of the pathogenic microbial organism, and the immunological response and the development of disease symptoms of the test animal or human may be evaluated. In the event the infection results in the development of an improved immune response, or less severe or no disease symptoms in the infected test animal or human than in an unvaccinated test animal or human exposed to the same infection, the vaccine formulation can be said to be cross-protective.

[0121] The term “pharmaceutically acceptable”, as used herein, refers to materials, including carriers, diluents, or auxiliary agent that are compatible with other materials in a pharmaceutical formulation, including a vaccine formulation, and within the scope of reasonable medical judgement suitable for use in contact with humans without excessive toxicity, allergic response, irritation, or other adverse response commensurate with a reasonable risk / benefit ratio.

[0122] The terms “treating” and “treatment”, and the like, as used herein, are intended to mean obtaining a desirable physiological, pharmacological, or biological effect. The effect may result in the prevention ( / .e., prophylactic treatment), inhibition, attenuation, reversal of a sign, symptom or cause of a disorder, or disease, attributable to the disorder, or disease. Clinical evidence of the treatment may vary with the disorder, or disease, the human, and the selected treatment. In the context of the treatment of an indication a physiological effect667451850may include, for example, diminished vaginal discharge, diminished penile discharge, improved dysuria.

[0123] The term “pharyngitis”, as used herein, refers to the accepted medical definition of pharyngitis and includes any pharyngitis developed upon infection and colonization by N. gonorrhoeae of the pharyngeal tract.

[0124] The terms “disseminated gonococcal infection” or “DGI”, as used herein, refers to the accepted medical definition of DGI and includes any DGI developed upon infection and colonization by / V. gonorrhoeae including DGI- associated arthritis, tenosynovitis, endocarditis and / or dermatitis.

[0125] The term “dysuria”, as used herein, refers to the accepted medical definition of dysuria and includes any dysuria state developed upon infection and colonization by N. gonorrhoeae of the female or male urinary tract.

[0126] The term “abnormal penile discharge”, as used herein refers to the accepted medical definition of abnormal penile discharge and includes any abnormal penile discharge developed upon infection and colonization by N. gonorrhoeae of the male genital tract.

[0127] The term “abnormal vaginal discharge”, as used herein, refers to the accepted medical definition of abnormal vaginal discharge and includes any abnormal vaginal discharge developed upon infection and colonization by N. gonorrhoeae of the female genital tract.

[0128] The terms “vaccine” and “vaccine formulation”, as used herein, refer to a pharmaceutically acceptable preparation that may be administered to a human to induce a humoral immune response (including eliciting a soluble antibody response) and / or cell-mediated immune response (including eliciting a cytotoxic T lymphocyte (CTL) response).

[0129] The terms “substantially pure” and “isolated”, as may be used interchangeably herein describe a compound, e.g., a polypeptide, which has been separated from components that naturally accompany it. Typically, a compound is substantially pure when at least 60%, more preferably at least 75%, more preferably at least 90%, 95%, 96%, 97%, or 98%, and most preferably at least 99% of the total material (by volume, by wet or dry weight, or by mole percent or mole fraction) in a sample is the compound of interest. Purity can be measured by any appropriate method, e.g., in the case of polypeptides, by chromatography, gel electrophoresis or HPLC analysis.677451850General implementation

[0130] As hereinbefore mentioned, the present disclosure relates to vaccine formulations. In general, the herein provided methods and compositions can be used to prevent, treat, or ameliorate Neisseria gonorrhoeae infections in humans. In this respect, the methods, and compositions of the present disclosure, in particular, may be used to prevent, treat, or ameliorate dysuria, pharyngitis, abnormal penile discharge, abnormal vaginal discharge, disseminated gonococcal infection (DGI), including DGI-associated arthritis, tenosynovitis, endocarditis and / or dermatitis, associated with Neisseria gonorrhoeae infections. The vaccine formulations of the present disclosure can provide long-term, effective protection against N. gonorrhoeae infections.

[0131] The inventors have discovered that the vaccine formulations of the present disclosure are cross-protective and can provide protection against infection by multiple Neisseria gonorrhoeae strains. Surprisingly, the vaccine formulations of the present disclosure can be cross-protective using only two immunogenic active agents.

[0132] Furthermore, the vaccine formulations of the present disclosure involve the use of polypeptide-based immunogenic active agents, and as such, the compositions and methods of the present disclosure do not involve the use of live attenuated microbial species, and thus avoid infection risks associated with the use of live vaccines.

[0133] Furthermore, the vaccine formulations of the present disclosure can limit the administration of antibiotics to humans, and thus limit the development of antibiotic resistant N. gonorrhoeae strains.

[0134] Furthermore, the polypeptide-based immunogenic active agents included in the vaccine formulations of the present disclosure may be prepared using a convenient recombinant production system, and the immunogenic active agents may be stably stored.

[0135] The present inventors have discovered, in particular, that proteins selected from a class of proteins known as TbpB proteins may be used as immunogenic active agents in the formulation of vaccines to prevent, treat or ameliorate Neisseria gonorrhoeae infections in humans. The TbpB proteins can be selected and obtained from N. gonorrhoeae strains. Preferably, the vaccines of687451850the present disclosure comprise two or at least two TbpB polypeptides, or equivalent immunogenic portions thereof, wherein at least one TbpB protein is an Ngo1 TbpB protein, and at least one TbpB protein is an Ngo2 TbpB protein. However, the TbpB proteins do not need to be selected from the same strain as the infecting N. gonorrhoeae strain, or even from the same phylogenetic cluster as the infecting N. gonorrhoeae strain.

[0136] In what follows example embodiments of the compositions and methods of the present disclosure are described.

[0137] Thus, the present disclosure provides, in at least one aspect, and in at least one embodiment, a vaccine formulation for the prevention, treatment or amelioration of N. gonorrhoeae infection in a human in need thereof, the vaccine formulation comprising an effective amount of at least two TbpB proteins or immunogenically equivalent portions thereof, or the vaccine formulation comprising an effective amount of at least two mRNA polynucleotides encoding the two N. gonorrhoeae TbpB proteins, or immunogenically equivalent portions thereof, wherein at least one TbpB protein is an Ngo1 TbpB protein ( / .e., selected from an Ngo1 phylogenetic cluster), and at least one TbpB protein is an Ngo2 TbpB protein ( / .e., selected from an Ngo2 phylogenetic cluster).

[0138] In another aspect, the present disclosure further provides, in at least one other embodiment, a method for the prevention, treatment or amelioration N. gonorrhoeae infection in a human in need thereof, the method comprising administering to the human a vaccine formulation comprising (i) at least two TbpB proteins, or immunogenically equivalent portions thereof, at least two mRNA polynucleotides encoding at least two N. gonorrhoeae TbpB proteins, wherein at least one TbpB protein is an Ngo1 TbpB protein, and at least one TbpB protein is an Ngo2 TbpB protein, and wherein the vaccine formulation is administered in an effective amount to human in need thereof to prevent, treat, or ameliorate the N. gonorrhoeae infection in a human.

[0139] In general, according to an aspect of the present disclosure, vaccine formulations comprising selected at least two TbpB proteins, or immunogenically equivalent portions thereof, or at least two mRNAs encoding at least two TbpB proteins, or immunologically equivalent portions thereof can be prepared or obtained. These formulations can be administered in effective amounts to a human in need thereof. Thus, in what follows next suitable preparations comprising697451850selected TbpB proteins, or immunogenically equivalent portions thereof, and mRNAs encoding selected TbpBs will be described, as well as methods of making selected TbpB proteins, or an immunogenically equivalent portion thereof, or mRNAs encoding selected mRNAs. Thereafter, vaccine formulations comprising at least two TbpB proteins, or immunogenically equivalent portions thereof, or at least two mRNAs encoding TbpB proteins, and methods of preparing such vaccine formulations and administering the same to a human in need thereof will be described.

[0140] Thus, initially, considering TbpB protein preparations, in an aspect of the present disclosure, preparations containing a selected TbpB protein, or an immunogenically equivalent portion thereof, can be prepared biosynthetically using a host cell system. In this respect, an isolated nucleic acid encoding an amino acid sequence corresponding with a TbpB protein or an immunogenically equivalent portion thereof can be introduced in host cells and expressed therein. It is noted that in what follows reference may be made to “TbpB protein”, “preparation containing a TbpB protein”, “formulation containing a TbpB protein”, or similar terms or expressions. When such reference is made, intended is for the same to be applicable to an Ngo1 TbpB protein and / or an Ngo2 TbpB protein, since the herein described preparations, formulations, methods, and techniques can generally be applied to make, obtain and / or use an Ngo1 TbpB protein or an Ngo2 TbpB protein without substantive adaptations. Where distinction between Ngo1 TbpB protein and Ngo2 TbpB protein is required or desired for context or clarity, reference is made to an “Ngo1 TbpB protein”, and / or an “Ngo2 TbpB protein”.

[0141] In general, any / V. gonorrhoeae TbpB protein may be used in accordance herewith. TbpB proteins can be obtained from N. gonorrhoeae bacteria, including any strain, variety, serogroup, or serotype of N. gonorrhoeae, including, in particular, strains belonging to phylogenetic cluster Ngo1 (Ngo1 TbpB) , or strains belonging to phylogenetic cluster Ngo2 (Ngo2 TbpB).

[0142] According to an aspect of the present disclosure, in an example embodiment, a nucleic acid sequence encoding an Ngo1 TbpB protein may be selected, wherein such nucleic acid includes SEQ.ID NO: 19, SEQ.ID NO: 21 , SEQ.ID and NO: 23, SEQ.ID which encode polypeptides having SEQ.ID NO: 20, SEQ.ID NO: 22, and SEQ.ID NO: 24, respectively.707451850

[0143] According to an aspect of the present disclosure, in an example embodiment, a nucleic acid sequence encoding an Ngo2 TbpB protein may be selected, wherein such nucleic acid includes SEQ.ID NO: 9 (encoding a polypeptide having SEQ.ID NO: 10), SEQ.ID NO: 31 , SEQ.ID NO: 32 (each encoding a polypeptide having SEQ.ID NO: 33), SEQ.ID NO: 36, SEQ.ID NO: 37 (each encoding a polypeptide having SEQ.ID NO: 38), SEQ.ID NO: 41 , SEQ.ID NO: 42 (each encoding a polypeptide having SEQ.ID NO: 43), SEQ.ID NO: 46, SEQ.ID NO: 47 (each encoding a polypeptide having SEQ.ID NO: 48), SEQ.ID NO: 51 , SEQ.ID NO: 52 (each encoding a polypeptide having SEQ.ID NO: 53), and SEQ.ID NO: 56, SEQ.ID NO: 57 (each encoding a polypeptide having SEQ.ID NO: 58).

[0144] In further example embodiments, the Ngo1 TbpB protein, or immunologically equivalent portion thereof, can be a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 1 , SEQ.ID NO: 5, SEQ.ID NO: 19, SEQ.ID and SEQ.ID. NO: 27;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 2, SEQ.ID NO: 6, SEQ.ID NO: 20, and SEQ.ID NO: 28, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 2, SEQ.ID NO: 6, SEQ.ID NO: 20, and SEQ.ID NO: 28, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e) or (f).717451850

[0145] In some embodiments, the Ngo2 TbpB protein, or an immunologically equivalent portion thereof, can be a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 9, SEQ.ID NO: 31 , SEQ.ID NO: 32, SEQ.ID NO: 36, SEQ.ID NO: 37, SEQ.ID NO: 41 , SEQ.ID NO: 42, SEQ.ID NO: 46, SEQ.ID NO: 47, SEQ.ID NO: 51 , SEQ.ID NO: 52, SEQ.ID NO: 56, and SEQ.ID NO: 57;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 33, SEQ.ID NO: 38 SEQ.ID NO: 43, SEQ.ID NO: 48, SEQ.ID NO: 53 and SEQ.ID NO: 58, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 33, SEQ.ID NO: 38 SEQ.ID NO: 43, SEQ.ID NO: 48, SEQ.ID NO: 53 and SEQ.ID NO: 58, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0146] TbpB proteins that may be used in accordance herewith include naturally occurring TbpB proteins, as well as TbpB proteins that may have been modified. Modifications in this respect include modifications made to the amino acid sequence of a TbpB polypeptide, including for example, a modification in one or more specific individual amino acids, which may be referred to as “site-directed mutations”, such as, for example, an Ngo2 TbpB protein having SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, or SEQ.ID NO: 100, which represent different site directed mutations of SEQ.ID NO: 10, as will be further described hereinafter.727451850

[0147] Particular modifications that according to certain embodiments hereof, may be made to TbpB proteins include modifications which diminish binding of the TbpB protein to a native host protein, (i.e., a protein in a human susceptible to N. gonorrhoeae infection). Such diminished binding can generally be assessed by evaluating the affinity of a TbpB protein for a native host protein. The affinity may be quantitatively evaluated by experimentally determining the dissociation constant (Kd) between the TbpB protein and the native host protein. Thus, for example, in this respect, the Kd between a native TbpB protein and a native host protein, for example, between TbpB and human host iron binding protein, may be compared with the Kd between a modified TbpB protein and the same native host protein. In general, the higher the Kd value, the weaker the affinity of the TbpB protein for the native host protein. In some embodiments, the Kd between a modified TbpB and a native host protein may exceed the Kd between the native non-modified TbpB and the native host protein by a factor of at least 2x, at least 5x, at least 10x, at least 25x, at least 50x, at least 10Ox, at least 250x, at least 500x, or at least 1000x. In some embodiments, the native nonmodified TbpB protein can comprise at least 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, or 25 consecutive amino acid residues which are identical to a modified TbpB protein, but for the modification of one, two, or three amino acid residues. Techniques to determine the Kd between two proteins are well known to those of skill in the art and include, for example isothermal calorimetry, surface plasmon resonance and biolayer interferometry (BLI) (see further, for example, Rich R. et al. 2007, Anal. Biochem. 361 : 1-6; Abdiche, Y. et al., 2008, Anal. Biochem. 377: 209-217; and Velazquez-Campoy, A. et al., 2004, Methods Mol. Biol. 261 : 35-54).

[0148] Examples of Ngo1 TbpB proteins that may be used for modification in this respect include TbpB proteins having SEQ.ID NO: 2, SEQ.ID NO: 6, SEQ.ID NO: 20, and SEQ.ID NO: 28 (encoded by nucleic acid sequences having SEQ.ID NO: 1 , SEQ.ID NO: 5, SEQ.ID NO: 19, and SEQ.ID NO: 27, respectively). In this respect, the Ngo1 TbpB polypeptides having SEQ.ID NO:26, SEQ.ID NO: 62, SEQ.ID NO: 64, SEQ.ID NO: 68, SEQ.ID NO:70, SEQ.ID NO: 72, SEQ.ID NO: 74, SEQ.ID NO: 76, SEQ.ID NO:80, SEQ.ID NO: 82, SEQ.ID NO: 84, SEQ.ID NO: 86, SEQ.ID NO: 88, SEQ.IDNO: 102, SEQ.ID NO: 108, SEQ.ID NO: 110, SEQ.ID NO: 116, SEQ.ID NO:737451850122, and SEQ.ID NO: 128, are modified and can exhibit reduced binding to human transferrin relative to native TbpBs.

[0149] Particular examples of Ngo1 proteins that are modified and that can exhibit reduced binding to human transferrin relative to native TbpBs include Ngo1 polypeptides comprising SEQ.ID NO: 26, SEQ.ID NO: 102, and SEQ.ID NO: 108.

[0150] Examples of Ngo2 TbpB proteins that may be used for modification in this respect include TbpB proteins having SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 32, SEQ.ID NO: 35, SEQ.ID NO: 37, SEQ.ID NO: 40, SEQ.ID NO: 42, SEQ.ID NO: 45, SEQ.ID NO: 47, SEQ.ID NO: 50, SEQ.ID NO: 52, SEQ.ID NO: 55, SEQ.ID NO: 57, and SEQ.ID NO: 60 (encoded by nucleic acid sequences having SEQ.ID NO: 9, SEQ.ID NO: 11 , SEQ.ID NO: 13, SEQ.ID NO: 31 , SEQ.ID NO: 34, SEQ.ID NO: 36, SEQ.ID NO: 39, SEQ.ID NO: 41 , SEQ.ID NO: 44, SEQ.ID NO: 46, SEQ.ID NO: 49, SEQ.ID NO: 51 , SEQ.ID NO: 54, SEQ.ID NO: 57, and SEQ.ID NO: 59, respectively). In this respect, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 48, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO: 216, SEQ.ID NO: 218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224, SEQ.ID NO: 226, SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232, SEQ.ID NO: 234, SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240, SEQ.ID NO: 242, SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248, SEQ.ID NO: 250, SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256, SEQ.ID NO: 258, SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264, SEQ.ID NO: 266, SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272, SEQ.ID NO: 274, SEQ.ID NO: 344, and SEQ.ID NO: 346, are modified and can exhibit reduced binding to human transferrin relative to native TbpBs.

[0151] Particular examples of Ngo2 proteins that are modified and that can exhibit reduced binding to human transferrin relative to native TbpBs include Ngo2 polypeptides comprising SEQ.ID NO: 230, SEQ.ID NO: 234, SEQ.ID NO: 238, and SEQ.ID NO: 346.

[0152] An example of an Ngo2 TbpB protein that may be used in this respect for modification includes the TbpB protein having SEQ.ID NO: 10 (encoded by the nucleic acid sequence having SEQ.ID NO: 9). In this respect, the Ngo2 TbpB747451850polypeptides having SEQ.ID 16, and SEQ.ID NO: 18 are modified to reduce binding to human transferrin relative to native TbpBs.

[0153] It is noted that when referring to modulated TbpB polypeptides herein, amino acid residues may be referred to by a specific number (e.g., aspartic acid amino acid residue 209 (or D209)). The numbering in conjunction with an amino acid residue in this respect corresponds with the sequential location of the amino acid residue in the amino acid sequence constituting a certain TbpB polypeptide. In this respect, pursuant to amino acid numbering practices conventionally used in the art, the N-terminal amino residue of the amino acid sequence constituting a polypeptide is referred to as amino acid number 1 , and the following amino acid residues are further sequentially numbered into C-terminal direction. Thus, for example, aspartic acid amino acid residue 209 (or D209) refers to the 209thamino acid residue from the N-terminal end of the amino acid sequence being an aspartic acid residue. The numbering in conjunction with specific amino acid residues further can refer to specific example native reference TbpB polypeptides, notably SEQ.ID NO: 2, SEQ.ID NO: 4, SEQ.ID NO: 6, SEQ.ID NO: 8, SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 28 SEQ.ID NO: 32, SEQ.ID NO: 35, SEQ.ID NO: 37, SEQ.ID NO: 40, SEQ.ID NO: 42, SEQ.ID NO: 45, SEQ.ID NO: 50, SEQ.ID NO: 52, SEQ.ID NO: 55, SEQ.ID NO: 57 and SEQ.ID NO: 60 (encoded by nucleic acid sequences having SEQ.ID NO: 1 , SEQ.ID NO: 3, SEQ.ID NO: 5, SEQ.ID NO: 7, SEQ.ID NO: 9, SEQ.ID NO: 11 , SEQ.ID NO: 13, SEQ.ID NO: 19, SEQ.ID NO: 21 , SEQ.ID NO: 27 SEQ.ID NO: 31 , SEQ.ID NO: 34, SEQ.ID NO: 36, SEQ.ID NO: 39, SEQ.ID NO: 41 , SEQ.ID NO: 44, SEQ.ID NO: 49, SEQ.ID NO: 51 , SEQ.ID NO: 54, SEQ.ID NO: 56 and SEQ.ID NO: 59, respectively).

[0154] Thus, for example, SEQ.ID NO: 22 can serve as a native reference polypeptide. Amino acid residue 209 in SEQ.ID NO: 22 corresponds with an aspartic acid amino acid residue (D209). SEQ.ID NO: 26 represents a modulated TbpB polypeptide in which aspartic acid residue D209 in SEQ.ID NO: 22 is replaced with a histidine residue. The modulated polypeptide may be referred to herein as “D209H”. Thus, it will be clear that the term “modulated amino acid residue”, as used herein, refers to an amino acid residue within a polypeptide sequence that differs from an amino acid residue found in the same position in another, similar polypeptide sequence, for example, a polypeptide that contains at757451850least 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, or 25 consecutive identical amino acid residues, as the polypeptide containing the modulated amino acid sequence, wherein within at least 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, or 25 consecutive identical amino acid residues, an amino acid residue is replaced with another amino acid residue. Furthermore, it will be understood that when considering the aforementioned two polypeptides, the amino acid numbering may not be identical. By way of example, in a certain polypeptide that is identical to SEQ.ID NO: 22, except that it lacks the first five N-terminal amino acid residues (and hence comprising at least 100 consecutive amino acids which are identical to SEQ.ID NO: 22), a D209H modulation will correspond with D204 being modulated in that certain polypeptide, however the amino acid residue being modulated can be said to be the same or corresponding amino acid residue. By way of a further example, amino acid residue 209 in SEQ.ID NO: 22 can be said to correspond with amino residue 231 in SEQ.ID NO: 26, as a result of SEQ.ID NO: 26 comprising a signal peptide extending the N-terminal end of the polypeptide. It is noted that SEQ.ID NO: 22 and SEQ.ID NO: 26 comprise at least 100 consecutive amino acids which are identical, but for a single modulated amino acid residue, namely D209H.

[0155] Thus, for example, referring to an example native TbpB polypeptide having amino acid sequence SEQ.ID NO: 20 encoded by SEQ.ID NO; 19, it is noted that SEQ.ID NO: 19 can be codon optimized and its signal peptide can be removed and replaced with another signal peptide. In this respect, SEQ.ID NO: 19 can yield codon optimized SEQ.ID NO: 21 , encoding polypeptide SEQ.ID NO: 22. Amino acid residue 209 corresponds with an aspartic acid amino acid residue (D209) in codon optimized sequence SEQ.ID NO: 22. SEQ.ID NO: 26 represents a modulated TbpB polypeptide in which aspartic acid residue D209 in SEQ.ID NO: 22 is replaced with a histidine residue. The modulated polypeptide may be referred to herein as D209H and, as noted, SEQ.ID NO: 22 and SEQ.ID NO: 26 share at least 100 consecutive amino acid residues, one of which (D209) is modulated.

[0156] In some embodiments, in order to modulate a TbpB polypeptide, a charged (acidic or basic) amino acid (Arg (R), Lys (K), His (H), Asp (D), Glu (E)), may be replaced with an uncharged (neutral) amino acid residue (Ser (S), Thr (T), Asn (N), Gin (Q), Cys (C), Gly (G), Pro (P), Ala (A), Vai (V), lie (I), Leu (L), Met (M), Phe (F), Tyr (Y), or Trp (W)).767451850

[0157] In some embodiments, in order to modulate a TbpB polypeptide, an uncharged (neutral) amino acid residue (Ser (S), Thr (T), Asn (N), Gin (Q), Cys (C), Gly (G), Pro (P), Ala (A), Vai (V), lie (I), Leu (L), Met (M), Phe (F), Tyr (Y), or Trp (W)) may be replace with a charged (acidic or basic) amino acid (Arg (R), Lys (K), His (H), Asp (D), Glu (E)).

[0158] In some embodiments, in order to modulate a TbpB polypeptide, a positively charged (basic) amino acid residue (Arg (R), Lys (K), His (H)), may be replaced with a negatively charged (acidic) amino acid residue (Asp (D), Glu (E)).

[0159] In some embodiments, in order to modulate a TbpB polypeptide, a negatively charged (acidic) amino acid residue (Asp (D), Glu (E)) may be replaced with a positively charged (basic) amino acid residue (Arg (R), Lys (K), His (H)).

[0160] In one particular embodiment, an TbpB protein can be a modulated TbpB protein, such that the Kd between the modulated TbpB protein and a native host iron binding protein exceeds the Kd between the native non-modified TbpB protein and the native host iron binding protein by a factor of at least 2x, at least 5x, at least 10x, at least 25x, at least 50x, at least 100x, at least 250x, at least 500x, or at least 1000x wherein, optionally the native non-modified TbpB protein can comprise at least 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, or 25 consecutive amino acid residues which are identical to the modulated TbpB protein, but for the modulation of one, two or three modulated amino acids comprised within such 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, or 25 consecutive amino acid residues, i.e., the one, two, or three modulated amino acid residues may be amino acid residues independently positioned from one another along the peptide chain of the at least 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, or 25 consecutive amino acid residues, and the one, two, or three modulated amino acid residues may be positioned anywhere along the polypeptide chain of the at least 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, or25 consecutive amino acid residues, e.g., in a central portion thereof (e.g., less than 50, 40, 30, 20 10, or 5 amino acids from the central one or two amino acids of the polypeptide chain), or towards the N-terminal portions or C- terminal portions thereof (for example, 5, 10, 15, 20, 25 amino acid residues from the N-terminal or C-terminal amino acid residue of the polypeptide chain).

[0161] In one particular embodiment, the Ngo1 TbpB protein can be a modulated Ngo1 TbpB protein, such that the Kd between the modulated Ngo1 TbpB protein and a native host iron binding protein exceeds the Kd between the777451850native non-modified Ngo1 TbpB protein and the native host iron binding protein by a factor of at least 2x, at least 5x, at least 10x, at least 25x, at least 50x, at least 100x, at least 250x, at least 500x, or at least 1000x wherein the native nonmodified Ngo1 TbpB protein can comprise at least 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, or 25 consecutive amino acid residues of SEQ.ID NO: 22, and wherein the modulated TbpB protein can comprise at least 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, or 25 consecutive amino acids identical thereto but for one, two, or three modulated amino acid residues within the at least 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, or 25 consecutive amino acid residues.

[0162] In one further particular embodiment, the modulated TbpB protein can comprise at least 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, or 25 consecutive amino acids identical thereto but for one modulated amino acid residue within the at least 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, or 25 consecutive amino acid residues, wherein the modulated amino acid residue corresponds with aspartic acid amino acid residue 209 (209D) in SEQ.ID NO: 22, and wherein, optionally, aspartic acid amino acid residue 209 (209D) is modulated to replace aspartic acid amino acid residue 209 (209D) with a positively charged amino acid residue, optionally, a histidine (H) amino acid residue.

[0163] In one further particular embodiment, in an aspect, the modulated TbpB protein can comprise at least 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, or 25 consecutive amino acids identical thereto but for one modulated amino acid residue within the at least 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, or 25 consecutive amino acid residues, wherein the modulated amino acid residue corresponds with aspartic acid amino acid residue 209 (209D) in SEQ.ID NO: 22, and wherein, and wherein the modulated TbpB sequence comprises SEQ.ID NO: 26.

[0164] In further particular example embodiments, involving modified TbpB proteins, TbpB proteins that may be used in this respect, the Ngo 1 TbpB protein can be a TbpB protein having SEQ.ID NO: 26, and the Ngo2 TbpB protein can be a TbpB protein having SEQ.ID NO: 16 or SEQ.ID NO: 18, as further detailed in Example 5.

[0165] According to an aspect of the present disclosure, suitable nucleic acid sequences include nucleic acid sequences encoding an immunogenically equivalent portion of a TbpB polypeptide, notably, in particular, TbpB polypeptide787451850portions which are at least immunologically equivalent to full length TbpB proteins, including the Ngo1 TbpB proteins set forth in SEQ.ID NO: 2, SEQ.ID NO: 6, SEQ.ID NO: 20, and SEQ.ID NO: 28, and the Ngo2 TbpB proteins set forth in SEQ.ID NO: 10, SEQ.ID NO: 33, SEQ.ID NO: 38 SEQ.ID NO: 43, SEQ.ID NO: 48, SEQ.ID NO: 53 and SEQ.ID NO: 58. Thus, further example nucleic acid sequences that may be used in selected embodiments include nucleic acid sequences encoding an amino acid sequence which is at least immunologically equivalent to a TbpB polypeptide, the amino acid sequence corresponding with at least 10 consecutive amino acids and up to 150 amino acids, including 10, 20, 30, 40, 50, 55, 60, 70, 80, 90, 100, 110, 120, 130, 140, or 150 amino acids of a full length TbpB protein, including the Ngo 1 TbpB protein set forth in in SEQ.ID NO: 2, SEQ.ID NO: 6, SEQ.ID NO: 20, and SEQ.ID NO: 28, and the Ngo2 TbpB protein set forth in SEQ.ID NO: 10, SEQ.ID NO: 33, SEQ.ID NO: 38 SEQ.ID NO: 43, SEQ.ID NO: 48, SEQ.ID NO: 53 and SEQ.ID NO: 58.

[0166] In a further embodiment, the TbpB protein, or immunogenically equivalent portion, thereof may be linked to another polypeptide to form a hybrid protein, including, for example, a carrier protein or extension to facilitate detection or purification, such as a polyhistidine extension (HIS-tag) or a FLAG-tag peptide extension, or another immunogenic polypeptide, including, further a second TbpB protein, or an immunogenically equivalent portion thereof, and further including a scaffold protein, for example, a TbpA polypeptide or portion thereof, in which a TbpB protein, or (one or more) immunogenically equivalent portions thereof, has been included.

[0167] Further examples of hybrid polypeptides included herein, are hybrid proteins comprising a first TbpB polypeptide fused to a second TbpB polypeptide, including an Ngo1 TbpB protein, or an immunogenic portion thereof, fused to an Ngo2 TbpB protein, or an immunogenic portion thereof, including, in particular, an Ngo 1 TbpB protein set forth in in SEQ.ID NO: 2, SEQ.ID NO: 6, SEQ.ID NO: 20, and SEQ.ID NO: 28, or an immunogenic portion thereof, fused to an Ngo2 TbpB protein set forth in SEQ.ID NO: 10, SEQ.ID NO: 33, SEQ.ID NO: 38 SEQ.ID NO: 43, SEQ.ID NO: 48, SEQ.ID NO: 53 and SEQ.ID NO: 58, or an immunogenic portion thereof.

[0168] Thus, it will now be clear that in accordance with the teachings herein various Ngo1 TbpB and Ngo2 TbpB polypeptides may be selected, including797451850native Ngo1 TbpB and Ngo2 TbpB polypeptides, and modulated Ngo1 TbpB and Ngo2 TbpB polypeptides. These Ngo1 TbpB and Ngo2 TbpB polypeptides include, at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 1 , SEQ.ID NO: 3, SEQ.ID NO: 5, SEQ.ID NO: 7, SEQ.ID NO: 19, SEQ.ID NO: 21 , SEQ.ID NO: 23, SEQ.ID NO: 25, SEQ.ID NO: 27, SEQ.ID NO: 29, SEQ.ID NO: 61 , SEQ.ID NO: 63, SEQ.ID NO: 65, SEQ.ID NO: 67, SEQ.ID NO: 69, SEQ.ID NO: 71 , SEQ.ID NO: 73, SEQ.ID NO: 75, SEQ.ID NO: 77, SEQ.ID NO: 79, SEQ.ID NO: 81 , SEQ.ID NO: 83, SEQ.ID NO: 85, SEQ.ID NO: 87, SEQ.ID NO: 101 , SEQ.ID NO: 103, SEQ.ID NO: 105, SEQ.ID NO: 107, SEQ.ID NO: 109, SEQ.ID NO: 111 , SEQ.ID NO: 113, SEQ.ID NO: 115, SEQ.ID NO: 117, SEQ.ID NO: 119, SEQ.ID NO: 121 , SEQ.ID NO: 123, SEQ.ID NO: 125, or SEQ.ID NO: 127;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 2, SEQ.ID NO: 4, SEQ.ID NO: 6, SEQ.ID. NO: 8, SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, SEQ.ID NO: 26, SEQ.ID NO: 28, SEQ.ID NO: 30, SEQ.ID NO: 62, SEQ.ID NO: 64, SEQ.ID NO: 66, SEQ.ID NO: 68, SEQ.ID NO: 70, SEQ.ID NO: 72, SEQ.ID NO: 74, SEQ.ID NO: 76, SEQ.ID NO: 78, SEQ.ID NO: 80, SEQ.ID NO: 82, SEQ.ID NO: 84, SEQ.ID NO: 86, SEQ.ID NO: 88, SEQ.ID NO: 102, SEQ.ID NO: 104, SEQ.ID NO: 106, SEQ.ID NO: 108, SEQ.ID NO: 110, SEQ.ID NO: 112 SEQ.ID NO: 114, SEQ.ID NO: 116, SEQ.ID NO: 187, SEQ.ID NO: 120, SEQ.ID NO: 122, SEQ.ID NO: 124, SEQ.ID NO: 126, or SEQ.ID NO: 128, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 2, SEQ.ID NO: 4,807451850SEQ.ID NO: 6, SEQ.ID. NO: 8, SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, SEQ.ID NO: 26, SEQ.ID NO: 28, SEQ.ID NO: 30, SEQ.ID NO: 62, SEQ.ID NO: 64, SEQ.ID NO: 66, SEQ.ID NO: 68, SEQ.ID NO: 70, SEQ.ID NO: 72, SEQ.ID NO: 74, SEQ.ID NO: 76, SEQ.ID NO: 78, SEQ.ID NO: 80, SEQ.ID NO: 82, SEQ.ID NO: 84, SEQ.ID NO: 86, SEQ.ID NO: 88, SEQ.ID NO: 102, SEQ.ID NO: 104, SEQ.ID NO: 106, SEQ.ID NO: 108, SEQ.ID NO: 110, SEQ.ID NO: 112, SEQ.ID NO: 114, SEQ.ID NO: 116, SEQ.ID NO: 187, SEQ.ID NO: 120, SEQ.ID NO: 122, SEQ.ID NO: 124, SEQ.ID NO: 126, or SEQ.ID NO: 128, or an immunogenically equivalent portion thereto; and (g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f); and at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 9, SEQ.ID NO: 11 , SEQ.ID NO: 13, SEQ.ID NO: 15, SEQ.ID NO: 17, SEQ.ID NO: 31 , SEQ.ID NO: 32, SEQ.ID NO: 34, SEQ.ID NO: 36, SEQ.ID NO: 37, SEQ.ID NO: 39, SEQ.ID NO: 41 , SEQ.ID NO: 42, SEQ.IDNO: 44, SEQ.ID NO: 46, SEQ.ID NO: 47, SEQ.ID NO: 49, SEQ.ID NO: 51 ,SEQ.ID NO: 52, SEQ.ID NO: 54, SEQ.ID NO: 56, SEQ.ID NO: 57, SEQ.IDNO: 59, SEQ.ID NO: 89, SEQ.ID NO: 91 , SEQ.ID NO: 93, SEQ.ID NO: 95,SEQ.ID NO: 97, SEQ.ID NO: 99, SEQ.ID NO: 207, SEQ.ID NO: 209, SEQ.IDNO: 211 , SEQ.ID NO: 213, SEQ.ID NO: 215, SEQ.ID NO: 217, SEQ.ID NO:219, SEQ.ID NO: 221 , SEQ.ID NO: 223, SEQ.ID NO: 225, SEQ.ID NO: 227,SEQ.ID NO: 229, SEQ.ID NO: 231 , SEQ.ID NO: 233, SEQ.ID NO: 235,SEQ.ID NO: 237, SEQ.ID NO: 239, SEQ.ID NO: 241 , SEQ.ID NO: 243,SEQ.ID NO: 245, SEQ.ID NO: 247, SEQ.ID NO: 249, SEQ.ID NO: 251 ,SEQ.ID NO: 253, SEQ.ID NO: 255, SEQ.ID NO: 257, SEQ.ID NO: 259,SEQ.ID NO: 261 , SEQ.ID NO: 263, SEQ.ID NO: 265, SEQ.ID NO: 267,SEQ.ID NO: 269, SEQ.ID NO: 271 , SEQ.ID NO: 273, SEQ.ID NO: 343, or SEQ.ID NO: 345;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);817451850(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.ID NO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.IDNO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94,SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.IDNO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO: 216, SEQ.ID NO:218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224, SEQ.ID NO: 226,SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232, SEQ.ID NO: 234,SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240, SEQ.ID NO: 242,SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248, SEQ.ID NO: 250,SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256, SEQ.ID NO: 258,SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264, SEQ.ID NO: 266,SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272, SEQ.ID NO: 274, SEQ.ID NO: 344, or SEQ.ID NO: 3< 6, or an immunoc enically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.IDNO: 35, SEQ.ID NO: 38, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43,SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.IDNO: 55, SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92,SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.IDNO: 208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO:216, SEQ.ID NO: 218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224,SEQ.ID NO: 226, SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232,SEQ.ID NO: 234, SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240,SEQ.ID NO: 242, SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248,827451850SEQ.ID NO: 250, SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256,SEQ.ID NO: 258, SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264,SEQ.ID NO: 266, SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272,SEQ.ID NO: 274, SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0169] In particular example embodiments, the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, can be a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 25;(b) a nucleic acid sequence having at least 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with the nucleic acid sequence of (a);(c) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to the nucleic acid sequence of (a);(e) a nucleic acid sequence encoding a polypeptide having the amino acid sequences set forth in SEQ.ID NO: 26, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of the amino acid sequence set forth in SEQ.ID NO: 26, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f); and the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, can be a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 9, SEQ.ID NO: 11 , SEQ.ID NO: 13, SEQ.ID NO: 15, SEQ.ID NO: 17, SEQ.ID NO: 31 , SEQ.ID NO: 32, SEQ.ID NO: 34, SEQ.ID NO: 36, SEQ.ID NO: 37, SEQ.ID NO: 39, SEQ.ID NO: 41 , SEQ.ID NO: 42, SEQ.ID NO: 44, SEQ.ID NO: 46, SEQ.ID NO: 47, SEQ.ID NO: 49, SEQ.ID NO: 51 ,837451850SEQ.ID NO: 52, SEQ.ID NO: 54, SEQ.ID NO: 56, SEQ.ID NO: 57, SEQ.ID NO: 59, SEQ.ID NO: 89, SEQ.ID NO: 91 , SEQ.ID NO: 93, SEQ.ID NO: 95,SEQ.ID NO: 97, SEQ.ID NO: 99, SEQ.ID NO: 207, SEQ.ID NO: 209, SEQ.IDNO: 211 , SEQ.ID NO: 213, SEQ.ID NO: 215, SEQ.ID NO: 217, SEQ.ID NO:219, SEQ.ID NO: 221 , SEQ.ID NO: 223, SEQ.ID NO: 225, SEQ.ID NO: 227,SEQ.ID NO: 229, SEQ.ID NO: 231 , SEQ.ID NO: 233, SEQ.ID NO: 235SEQ.ID NO: 237, SEQ.ID NO: 239, SEQ.ID NO: 241 , SEQ.ID NO: 243SEQ.ID NO: 245, SEQ.ID NO: 247, SEQ.ID NO: 249, SEQ.ID NO: 251SEQ.ID NO: 253, SEQ.ID NO: 255, SEQ.ID NO: 257, SEQ.ID NO: 259SEQ.ID NO: 261 , SEQ.ID NO: 263, SEQ.ID NO: 265, SEQ.ID NO: 267SEQ.ID NO: 269, SEQ.ID NO: 271 , SEQ.ID NO: 273, SEQ.ID NO: 343, or SEQ.ID NO: 345;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.ID NO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.IDNO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO: 216, SEQ.ID NO218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224, SEQ.ID NO: 226SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232, SEQ.ID NO: 234,SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240, SEQ.ID NO: 242,SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248, SEQ.ID NO: 250,SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256, SEQ.ID NO: 258,SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264, SEQ.ID NO: 266,SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272, SEQ.ID NO: 274,847451850SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.ID NO: 35, SEQ.ID NO: 38, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO:216, SEQ.ID NO: 218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224,SEQ.ID NO: 226, SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232,SEQ.ID NO: 234, SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240,SEQ.ID NO: 242, SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248,SEQ.ID NO: 250, SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256,SEQ.ID NO: 258, SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264,SEQ.ID NO: 266, SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272,SEQ.ID NO: 274, SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0170] In particular example embodiments, the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, can be a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 25;(b) a nucleic acid sequence having at least 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with the nucleic acid sequence of (a);(c) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to the nucleic acid sequence of (a);857451850(e) a nucleic acid sequence encoding a polypeptide having the amino acid sequences set forth in SEQ.ID NO: 26, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of the amino acid sequence set forth in SEQ.ID NO: 26, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f); and the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, can be a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 15, or SEQ.ID NO: 17;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 16 or SEQ.ID NO: 18, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 16 or SEQ.ID NO: 18, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0171] In further particular example embodiments, the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, included in a vaccine formulation can comprise:SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, or SEQ.ID NO: 26, or immunologically equivalent portion thereof;867451850and the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, included in a vaccine formulation can comprise: any of the amino acid sequences setforth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.ID NO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO:216, SEQ.ID NO: 218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO:224, SEQ.ID NO: 226, SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO:232, SEQ.ID NO: 234, SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO:240, SEQ.ID NO: 242, SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO:248, SEQ.ID NO: 250, SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO:256, SEQ.ID NO: 258, SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO:264, SEQ.ID NO: 266, SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO:272, SEQ.ID NO: 274, SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereof.

[0172] In further example embodiments, the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, included in a vaccine formulation can comprise:SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, orSEQ.ID NO: 26, or immunologically equivalent portion thereof; and the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, included in a vaccine formulation can comprise: any of the amino acid sequences setforth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, or SEQ.ID NO: 100, or an immunogenically equivalent portion thereof.

[0173] The thus selected nucleic acid sequences encoding a TbpB polypeptide may be expressed in a host cell to thereby produce the TbpB polypeptide. As is known to those of skill in the art, generally expression of nucleic877451850acids in a host cell, to produce a protein thereby biosynthetically, can be achieved by providing one or more nucleic acids capable of controlling expression in a host cell, and operably linking the one or more nucleic acids capable of controlling expression in a host cell to the nucleic acid one wishes to express. Such operable linking of a nucleic acid controlling expression generally involves linking in the 5’ to 3’ direction of expression the nucleic acid capable of controlling expression in a host cell to the nucleic acid one wishes to express, i.e., within the context of the instant disclosure, a TbpB protein. Nucleic acid sequences capable of controlling expression in host cells that may be used herein include any transcriptional promoter capable of controlling expression of polypeptides in host cells. Generally, promoters obtained from bacterial cells are used when a bacterial host cell is selected, while a yeast promoter will be used when a yeast host cell is selected, a plant promoter will be used when a plant cell is selected, and so on. The obtained nucleic acid comprising a promoter and the nucleic acid expressing a TbpB protein is generally a chimeric nucleic acid. Further nucleic acid elements capable elements of controlling expression in a host cell include transcriptional terminators, enhancers and the like, all of which may be included in the chimeric nucleic acid sequences of the present disclosure.

[0174] In accordance with the present disclosure, the chimeric nucleic acid sequences can be integrated into a recombinant expression vector which ensures good expression in the host cell, wherein the recombinant expression vector is suitable for expression in a host cell. The term “suitable for expression in a host cell” means that the recombinant expression vector comprises the chimeric nucleic acid linked to genetic elements required to achieve expression in a cell. As noted, such genetic elements can include transcriptional promoters, terminators, and enhancers, and the like. Further genetic elements that may be included in the expression vector are one or more nucleic acid sequences encoding marker genes, and one or more origins of replication. In some embodiments, the expression vector can freely replicate in the host cell. In other embodiments, the chimeric nucleic acid can be integrated into the host cell’s genomic DNA. In some embodiments, the expression vector further can comprise genetic elements required for the integration of the vector or a portion thereof in the host cell's genome, for example, if a plant host cell is used the T-DNA left and right border887451850sequences which facilitate the integration into the plant's nuclear genome can be included in the vector.

[0175] Marker genes that may be used in accordance with the present disclosure include all genes that allow the distinction of transformed cells from nontransformed cells, including all selectable and screenable marker genes. A marker gene may be a resistance marker such as an antibiotic resistance marker against, for example, kanamycin, chloramphenicol, methotrexate, or ampicillin. In other instances, a marker gene may be a gene which allows a cell to produce an essential nutrient, for example amino acids.

[0176] Thus, in an aspect, the present disclosure provides, in accordance with the teachings herein, in at least one embodiment, an expression vector comprising a chimeric nucleic acid sequence comprising as operably linked components:(a) a nucleic acid sequence encoding a N. gonorrhoeae TbpB protein, protein, or an immunogenically equivalent portion thereof; and(b) a nucleic acid sequence capable of controlling expression of the nucleic acid sequence encoding the N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof in a host cell, wherein the TbpB protein is an Ngo1 TbpB protein or an Ngo2 TbpB protein.

[0177] In a further aspect, the present disclosure provides, in accordance with the teachings herein, in at least one embodiment, an expression vector comprising a chimeric nucleic acid sequence comprising as operably linked components:(a) a nucleic acid sequence encoding a first and second N. gonorrhoeae TbpB protein, or immunogenically equivalent portions thereof; and(b) one or more nucleic acid sequences capable of separately or jointly controlling expression of the nucleic acid sequence encoding the first and second N. gonorrhoeae TbpB protein, or immunogenically equivalent portions thereof in a host cell, wherein the first and second TbpB protein are an Ngo1 TbpB protein and an Ngo2 TbpB protein, respectively.

[0178] Turning now to the host cell, it is noted, initially, that any host cell which upon cultivation expresses the chimeric nucleic acid can be selected and897451850used in accordance with the present disclosure. Suitable host cells in this respect include, for example, microbial cells, such as bacterial cells, yeast cells, for example, and algal cells or plant cells. A variety of techniques and methodologies to manipulate host cells to introduce nucleic acid sequences in cells and attain expression exists and are well known to the skilled artisan. These methods include, for example, cation-based methods, for example, lithium ion or calcium ion-based methods, electroporation, biolistics, and glass beads-based methods. As will be known to those of skill in the art, depending on the host cell selected, the methodology to introduce nucleic acid material in the host cell may vary, and, furthermore, methodologies may be optimized for uptake of nucleic acid material by the host cell, for example, by comparing uptake of nucleic acid material using different conditions. Detailed guidance can be found, for example, in Sambrook et al., Molecular Cloning, a Laboratory Manual, Cold Spring Harbor Laboratory Press, 2012, Fourth Ed. It is noted that the chimeric nucleic acid is a non-naturally occurring chimeric nucleic acid sequence and can be said to be heterologous to the host cell.

[0179] One example host cell that conveniently may be used is Escherichia coli. The preparation of the E. coli vectors may be accomplished using commonly known techniques such as restriction digestion, ligation, gel electrophoresis, DNA sequencing, the polymerase chain reaction (PCR) and other methodologies. A wide variety of cloning vectors is available to perform the necessary steps required to prepare a recombinant expression vector. Among the vectors with a replication system functional in E. coli, are vectors such as pBR322, the pUC series of vectors, the M13 mp series of vectors, pBluescript etc. Suitable promoter sequences for use in E. coli include, for example, the T7 promoter, the T5 promoter, tryptophan (trp) promoter, lactose (lac) promoter, tryptophan / lactose (tac) promoter, lipoprotein (Ipp) promoter, and A phage PL promoter. Typically, cloning vectors contain a marker, for example, an antibiotic resistance marker, such as ampicillin or kanamycin resistance marker, allowing selection of transformed cells. Nucleic acid sequences may be introduced in these vectors, and the vectors may be introduced in E. coli by preparing competent cells, electroporation or using other well-known methodologies to a person of skill in the art. E. coli may be grown in an appropriate medium, such as Luria-Broth medium907451850and harvested. Recombinant expression vectors may readily be recovered from cells upon harvesting and lysing of the cells.

[0180] Another example host cell that may be conveniently used is a yeast cell. Example yeast host cells that can be used are yeast cells belonging to the genus Candida, Kluyveromyces, Saccharomyces, Schizosaccharomyces, Pichia, Hansenula, and Yarrowia. In specific example embodiments, the yeast cell can be a Saccharomyces cerevisiae cell, a Yarrowia lipolytica cell, or Pichia pastoris cell.

[0181] A number of vectors exist for the expression of recombinant proteins in yeast host cells. Examples of vectors that may be used in yeast host cells include, for example, Yip type vectors, YEp type vectors, YRp type vectors, YCp type vectors, pGPD-2, pAO815, pGAPZ, pGAPZa, pHIL-D2, pHIL-S1 , pPIC3.5K, pPIC9K, pPICZ, pPICZa, pPIC3K, pHWO10, pPUZZLE and 2 pm plasmids. Such vectors are known to the art and are, for example, described in Cregg et al., Mol. Biotechnol. (2000) 16(1): 23-52. Suitable promoter sequences for use in yeast host cells are also known and described, for example, in Mattanovich et al., Methods Mol. Biol., 2012, 824:329-58, and in Romanos et al., 1992, Yeast 8: 423- 488. Examples of suitable promoters for use in yeast host cells include promoters of glycolytic enzymes, like triosephosphate isomerase (TPI), phosphoglycerate kinase (PGK), glyceraldehyde-3-phosphate dehydrogenase (GAPDH or GAP) and variants thereof, lactase (LAC) and galactosidase (GAL), P. pastoris glucose-6- phosphate isomerase promoter (PPGI), the 3-phosphoglycerate kinase promoter (PPGK), the glycerol aldehyde phosphate dehydrogenase promoter (PGAP), translation elongation factor promoter (PTEF), S. cerevisiae enolase (ENO-1 ), S. cerevisiae galactokinase (GAL1), S. cerevisiae alcohol dehydrogenase / glyceraldehyde-3-phosphate dehydrogenase (ADH1 , ADH2 / GAP), S. cerevisiae triose phosphate isomerase (TPI), S. cerevisiae metallothionein (CUP1), and S. cerevisiae 3-phosphoglycerate kinase (PGK), and the maltase gene promoter (MAL). Marker genes suitable for use in yeast host cells are also known to the art. Thus, antibiotic resistance markers, such as ampicillin resistance markers, can be used in yeast, as well as marker genes providing genetic functions for essential nutrients, for example, leucine (LEU2), tryptophan (TRP1 and TRP2), uracil (URA3, URA5, URA6), histidine (HIS3), and the like. Methods for introducing vectors into yeast host cells can, for example, be found in S. Kawai et al., 2010, Bioeng. Bugs 1 (6): 395-403.917451850

[0182] Yet other example host cells that may be used in accordance herewith are plant cells. Methods for introducing nucleic acids in plant cells are known to those of skill in the art. Agrobacterium mediated plant cell transformation methods are described, for example, by Gelvin S. in Microbiol. Mol. Biol. Rev., 2003, 67(1): 16-37, and physical transformation-based methods for plant cells are described by Rivera A.L. et al., 2012, Phys. Life Rev. 9(3): 308-345. Plant selectable marker genes are known to those of skill in the art and include antibiotic resistance genes, for example kanamycin resistance genes, and herbicide resistance genes, such as the bar and pat genes (Wohlleben et al., 1988, Gene 70:25-37). Screenable markers that may be employed to identify plant transformants through visual inspection include p-glucuronidase (GUS) (U.S. Pat. Nos. 5,268,463 and 5,599,670) and green fluorescent protein (GFP) (Niedz et al., 1995, Plant Cell Rep., 14: 403). Plant promoters are also known to those in the art and include, for example, constitutive promoters, such as the 35S cauliflower mosaic virus (CaMV) promoter (Rothstein et al., 1987, Gene 53: 153-161), the rice actin promoter (McElroy et al., 1990, Plant Cell 2:163-171 ; U.S. Pat. No. 6,429,357), a ubiquitin promoter, such as the corn ubiquitin promoter (U.S. Pat. Nos 5,879,903 and 5,273,894), and the parsley ubiquitin promoter (Kawalleck, P. et al., 1993, Plant Mol. Biol. 21 :673-684), and organ specific promoters, such as seed specific promoters, for example, a phaseolin promoter (Sengupta-Gopalan et al., 1985, Proc. Natl. Acad. Sci. USA 82: 3320-3324), or an oleosin promoter (U.S. Pat. No. 5,792,922).

[0183] Further, guidance with respect to the preparation of expression vectors and introduction thereof into host cells, including in E. coli cells, yeast cells, and other host cells, may be found in, for example: Sambrook et al., Molecular Cloning, a Laboratory Manual, Cold Spring Harbor Laboratory Press, 2012, Fourth Ed.

[0184] Thus, in another aspect, the present disclosure further provides, in at least one other embodiment, a host cell comprising a chimeric nucleic acid comprising as operably linked components:(a) a nucleic acid sequence encoding a N. gonorrhoeae TbpB protein, protein, or an immunogenically equivalent portion thereof; and927451850(b) a nucleic acid sequence capable of controlling expression of the nucleic acid sequence encoding the N. gonorrhoeae TbpB protein, or immunogenically equivalent portion thereof in a host cell, wherein the TbpB protein is an Ngo1 TbpB protein or an Ngo2 TbpB protein.

[0185] In an example embodiment, a single nucleic acid sequence encoding both an Ngo1 TbpB protein and an Ngo2 TbpB protein, optionally configured to produce an Ngo1 TbpB protein / Ngo2 TbpB fusion polypeptide, may be introduced in a host cell.

[0186] Thus, in another aspect, the present disclosure further provides, in at least one other embodiment, a host cell comprising a chimeric nucleic acid sequence comprising as operably linked components:(a) a nucleic acid sequence encoding a first and second N. gonorrhoeae TbpB protein, or immunogenically equivalent portions thereof; and(b) one or more nucleic acid sequences capable of separately or jointly controlling expression of the nucleic acid sequence encoding the first and second N. gonorrhoeae TbpB protein, or the immunogenically equivalent portions thereof in a host cell, wherein the first and second TbpB protein are selected from an Ngo1 TbpB protein, and an Ngo2 TbpB protein, respectively.

[0187] In an example embodiment, separate cells producing an Ngo1 TbpB protein and an Ngo2 TbpB protein may be prepared and such cells may be mixed. Such cells may be mixed to be grown together (as herein after further described), or they may be mixed following growth, for example, to separate an Ngo1 TbpB protein / Ngo2 TbpB protein mixture.

[0188] Thus, in another aspect, the present disclosure further provides, in at least one other embodiment, a mixture comprising a first and second host cell, the first host cell comprising a first chimeric nucleic acid comprising as operably linked components:(a) a nucleic acid sequence encoding a first N. gonorrhoeae TbpB protein, protein, or an immunogenically equivalent portion thereof; and(b) a nucleic acid sequence capable of controlling expression of the nucleic acid sequence encoding the first N. gonorrhoeae TbpB937451850protein, or immunogenically equivalent portion thereof, in the first host cell, wherein the first TbpB protein is an Ngo1 TbpB protein; and the second host cell comprising a second chimeric nucleic acid comprising as operably linked components:(c) a nucleic acid sequence encoding a second N. gonorrhoeae TbpB protein, protein, or an immunogenically equivalent portion thereof; and(d) a nucleic acid sequence capable of controlling expression of the nucleic acid sequence encoding the second N. gonorrhoeae TbpB protein, or immunogenically equivalent portion thereof, in the second host cell, wherein the second TbpB protein is an Ngo2 TbpB protein.

[0189] Thus, to briefly recap, a host cell comprising a chimeric nucleic acid comprising (i) a nucleic acid sequence encoding a first and / or second TbpB protein or an immunogenically equivalent portion thereof; and (ii) a nucleic acid sequence capable of controlling expression of the nucleic acid sequence encoding a first and / or TbpB protein, or an immunogenically equivalent portion thereof in a host cell, wherein the first and / or second TbpB protein is an Ngo1 TbpB protein and / or an Ngo2 protein, can be prepared in accordance with the present disclosure.

[0190] In accordance herewith, host cells can be grown to multiply and to express a chimeric nucleic acid. Expression of the chimeric nucleic acid results in the biosynthetic production in the host cell of a TbpB protein of an immunogenically equivalent portion thereof. Growth media and growth conditions can vary depending on the host cell that is selected, as will be readily appreciated to those of ordinary skill in the art. Growth media typically contain a carbon source, one or several nitrogen sources, essential salts including salts of potassium, sodium, magnesium, phosphate and sulphate, trace metals, water soluble vitamins, and process aids including but not limited to antifoam agents, protease inhibitors, stabilizers, ligands and inducers. Typical carbon sources are e.g., mono- or disaccharides. Typical nitrogen sources are, e.g., ammonia, urea, amino acids, yeast extract, corn steep liquor and fully or partially hydrolyzed proteins. Typical trace metals are e.g., Fe, Zn, Mn, Cu, Mo and H3BO3. Typical water-soluble947451850vitamins are e.g., biotin, pantothenate, niacin, thiamine, p- aminobenzoic acid, choline, pyridoxine, folic acid, riboflavin, and ascorbic acid. Further, specific example media include liquid culture media for the growth of yeast cells and bacterial cells including, Luria-Bertani (LB) broth for bacterial cell cultivation, and yeast extract peptone dextrose (YEPD or YPD), for yeast cell cultivation. Further media and growth conditions can be found in Sambrook et al., Molecular Cloning, a Laboratory Manual, Cold Spring Harbor Laboratory Press, 2012, Fourth Ed.

[0191] Upon production by the host cells of a TbpB protein or an immunogenically equivalent portion thereof, the TbpB protein or the immunogenically equivalent portion thereof may be recovered from the host cells, and separated from other constituents, such as cellular debris, or media constituents, for example. Separation techniques will be known to those of skill in the art and include a variety of different protein purification techniques including, e.g., ion-exchange chromatography, size exclusion chromatography, affinity chromatography, hydrophobic interaction chromatography, reverse phase chromatography, gel filtration, etc. Further general guidance with respect to protein purification may for example be found in: Cutler, P. Protein Purification Protocols, Humana Press, 2004, Second Ed. Thus, substantially pure preparations of TbpB proteins or immunogenically equivalent portions thereof may be obtained. The recovered TbpB proteins may be obtained in a more or less pure form, for example, a preparation of a TbpB protein or immunogenically equivalent portion thereof having a purity of at least about 60% (w / v), about 70% (w / v), about 80% (w / v), about 90% (w / v), about 95% (w / v), or about 99% (w / v) may be obtained.

[0192] Furthermore, it is noted that the recombinant production of the TbpB proteins or immunogenically equivalent portions thereof in host cell system permits the production thereof in a manner in which the TbpB proteins or immunogenically equivalent portions are substantially free from other N. gonorrhoeae constituent materials, such as other N. gonorrhoeae proteins, membrane materials, lipopolysaccharides, and the like, naturally associated with TbpB proteins.

[0193] It is noted that the cells, in some embodiments, may secrete a portion of the produced TbpB protein or immunogenically equivalent portion thereof the in the cell growth medium, thus a portion of the produced TbpB protein or immunogenically equivalent portion thereof may be recovered from the cells957451850and a further portion of the TbpB protein or immunogenically equivalent portion thereof may be recovered from the growth medium.

[0194] It is further noted that the vaccine formulations of the present disclosure may comprise TbpB proteins or an immunogenically equivalent portion thereof in more or less pure form. Thus, in accordance herewith, a substantially pure TbpB protein or immunogenically equivalent portion thereof may be obtained and used to prepare human vaccine formulations. Thus, for example, in some embodiments, the TbpB protein may be substantially free of other host cell constituent materials, such as host cell proteins, membrane materials, lipopolysaccharides, and the like. In other embodiments, more crude preparations comprising TbpB protein or immunogenically equivalent portion thereof may be obtained and used to prepare vaccine formulations. Thus, for example, in such embodiments, host cells, host cell lysates or host cell fractions comprising the TbpB protein or immunogenically equivalent portion thereof may be used to prepare the vaccine formulations.

[0195] It is noted that any TbpB or immunogenically equivalent portion thereof may be used to formulate the vaccine formulations of the present disclosure, including a TbpB protein having any one of the amino acid sequences set forth in SEQ.ID NO: 2, SEQ.ID NO: 4, SEQ.ID NO: 6, SEQ.ID. NO: 8 (Ngo1 TbpB proteins), SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14 SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID. NO: 20, SEQ.ID. NO: 22 (Ngo2 TbpB proteins), or an immunogenically equivalent portion thereof.

[0196] It is noted that a N. gonorrhoeae strain may be isolated from infected humans and cultured in order to identify the N. gonorrhoeae strain, and the sequence of the TbpB polypeptide present therein may be determined using techniques known to those of ordinary skill in the art.

[0197] Thus, it will now be clear that in one embodiment, the vaccine formulation can comprise at least two N. gonorrhoeae TbpB proteins, or immunogenically equivalent portions thereof, wherein the TbpB proteins at least one Ngo1 TbpB protein and at least one Ngo2 TbpB protein.

[0198] It is noted that the vaccine formulations of the present disclosure can be used to ameliorate or prevent infections by N. gonorrhoeae strains, other than the strains from which the TbpB protein included in a vaccine formulation is selected. Thus, the vaccine formulations of the present disclosure, surprisingly, do967451850not necessarily need to include TbpBs from a plurality of / V. gonorrhoeae strains in order to be used for the treatment of a human, even if the human can be or has been exposed to, or can be or has been infected by a plurality of N. gonorrhoeae strains. Thus, for example, a vaccine formulation comprising a TbpB protein obtained from a N. gonorrhoeae strain of a first strain, may be used to treat a human for an infection caused by a N. gonorrhoeae strain. In this respect, the vaccine formulations, even if they include only two TbpB polypeptides , or an immunogenically equivalent portion thereof, can be said to be cross-protective. Thus, for example, the vaccine formulations of the present disclosure can be cross-protective against infection by N. gonorrhoeae strains MS11 , FA1090, FA19, WHO F, WHO G, WHO L, WHO M, WHO O, WHO, P, WHO V, WHO W, WHO X, or WHO Y, without including a TbpB protein from any of these strains.

[0199] Notwithstanding the foregoing, in some embodiments, vaccine formulations may comprise three or more TbpB proteins.

[0200] Thus, to briefly recap, in accordance with the present disclosure N. gonorrhoeae TbpB polypeptides, or immunogenically equivalent portions thereof, may be selected for inclusion in vaccine formulations to treat humans infected by N. gonorrhoeae. Preferably, the vaccine formulations include at least two TbpB polypeptides selected N. gonorrhoeae TbpB polypeptide belonging to phylogenetic cluster Ngo1 and Ngo2, / .e., at least one Ngo1 TbpB protein and at least one Ngo2 TbpB protein.

[0201] In another embodiment, two mRNA polynucleotides encoding at least two N. gonorrhoeae TbpB proteins, may be prepared and included in a vaccine formulation, wherein the two mRNA polynucleotide encode at least one Ngo1 TbpB protein, and at least one Ngo2 TbpB protein, or immunologically equivalent portions thereof. Preferably, the vaccine formulations include at least two mRNAs encoding a TbpB polypeptide, one selected N. gonorrhoeae TbpB polypeptide belonging to phylogenetic cluster Ngo1 and one selected N. gonorrhoeae TbpB polypeptide belonging to phylogenetic cluster Ngo2.

[0202] It is noted that in embodiments wherein mRNA encoding an Ngo1 TbpB polypeptide, or an immunogenically equivalent portion thereof, or an Ngo2 TbpB polypeptide, or an immunogenically equivalent portion thereof, is produced, the mRNA encoding an Ngo1 TbpB polypeptide or Ngo2 TbpB polypeptide, may be produced in vivo, or in vitro. In vitro methods for synthesizing mRNA may977451850include, for example, an in vitro transcription (IVT) system, using a template polynucleotide, for example, a template cDNA or linearized plasmid DNA, encoding an Ngo1 TbpB polypeptide or Ngo2 TbpB polypeptide, or an immunogenically equivalent portion thereof. IVT systems typically include a transcription buffer, nucleotide triphosphates (NTPs), and RNAse inhibitor, and an RNA-polymerase. An example IVT system that may be used in this respect is described in PCT / US2014 / 026835. The mRNAs may be non-chemically modified and contain the conventional naturally occurring ribonucleotides, / .e., adenosine, guanosine, cytosine, and uridine. In other embodiments, the mRNAs may comprise chemically modified ribonucleotides, e.g., 1-methyl-pseudouridine (m1 i ), 1-ethyl-pseudouridine (e1 1), 5-methoxy-uridine (mo5U), 5-methyl-cytidine (m5C), and / or pseudouridine ( 1), 5-methoxym ethyl uridine, 5-methylthio uridine, 1 -methoxymethyl pseudouridine, 5-methyl cytidine, and / or 5-methoxy cytidine. When using an IVT system, non-modified and chemically modified NTPs may be included in the IVT system.

[0203] In some embodiments, the template can include a 5’-untranslated region (5’-UTR) ( / .e., the region that is located immediately upstream (5’) of the start codon, / .e., the first codon of an mRNA translated by a ribosome) and / or a 3’- untranslated region (3’-UTR) ( / .e., the region that is located immediately downstream (3’) of the stop codon, / .e., the last codon of an mRNA translated by a ribosome). The 5’-UTR and 3’-UTR do not encode protein, however they may contain stabilizing elements, such as a 3’-polyA tail, or a 5’-terminal cap.

[0204] The 5’-UTR may contain a promoter sequence, for example, a T7- promoter sequence, or any other suitable promoter sequence. The 5’-UTR may further contain a 5’ -terminal cap, for example, a 3'-0-Me-m7G(5')ppp(5') G (ARCA cap; New England BioLabs, Ipswich, Mass., USA), which may be included concomitantly with in vitro transcription. The 3’-UTR may contain a polyA tail, including, for example, 10 - 300 monophosphates, or it may contain AU rich elements (AREs). 5’-UTRs and 3’-UTRs may be heterologous to the mRNA sequence being expressed or synthetic ( / .e., not occurring in nature), including natural sequences that have been mutated. Example 5'-UTRs include Xenopus or human derived a-globin or p-globin (see: U.S. Pat. No. 8,278,036; 9,012,219), human cytochrome b-245a polypeptide, and hydroxysteroid (17b)987451850dehydrogenase, and Tobacco etch virus (see: U.S. Pat. Nos. 8,278,036, 9,012,219). Example 3’-UTRs include including Xenopus p-globin UTRs and human p -globin UTRs (see: U.S. Pat. Nos. 8,278,036; 9,012,219; US20110086907). A further modified p-globin construct which provides enhanced stability in some cell types by cloning two sequential human p-globin 3'-UTRs head to tail that may be used is well known in the art (see: Holtkamp, S. et al. , 2006, Blood, 109 (13), 4009 - 4017; WO2017 / 060314). Those of ordinary skill in the art will understand that 5'-UTRs that are heterologous or synthetic may be used with any desired 3'-UTR sequence. For example, a heterologous 5-'UTR may be used with a synthetic 3'-UTR with or with heterologous 3'-UTR.

[0205] Purification of the nucleic acids described herein may include, but is not limited to, nucleic acid clean-up, quality assurance and quality control. Cleanup may be performed by methods known in the arts such as, but not limited to, AGENCOURT® beads (Beckman Coulter Genomics, Danvers, Mass., USA), poly- T beads, LNA™ oligo-T capture probes (EXIQON® Inc, Vedbaek, Denmark) or F1 PLC based purification methods such as, but not limited to, strong anion exchange F1 PLC, weak anion exchange F1 PLC, reverse phase F1 PLC (RP- F1 PLC), and hydrophobic interaction F1 PLC (F1 IC-F1 PLC).

[0206] Thus, in another aspect, the present disclosure further provides in one embodiment expression vectors and host cells expressing mRNAs encoding TbpB proteins.

[0207] The present disclosure provides, in one embodiment, an expression vector comprising a chimeric nucleic acid sequence comprising as operably linked components:(a) a nucleic acid sequence encoding an mRNA encoding a first N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof; and(b) a nucleic acid sequence capable of controlling expression of the nucleic acid sequence encoding the mRNA encoding a first N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, in a host cell, wherein the TbpB protein is an Ngo1 TbpB protein, or an Ngo2 TbpB protein.997451850

[0208] In another embodiment, the present disclosure provides an expression vector comprising a chimeric nucleic acid sequence comprising as operably linked components:(a) a nucleic acid sequence encoding an mRNA encoding a first and second N. gonorrhoeae TbpB protein, or immunogenically equivalent portions thereof; and(b) one or more nucleic acid sequences capable of separately or jointly controlling expression of the nucleic acid sequence the mRNA encoding the first and second N. gonorrhoeae TbpB protein, or immunogenically equivalent portions thereof, in a host cell, wherein the first and second TbpB protein are an Ngo1 TbpB protein, and an Ngo2 TbpB protein, respectively.

[0209] The present disclosure provides, in one embodiment, a host cell comprising a chimeric nucleic acid comprising as operably linked components:(a) a nucleic acid sequence encoding a N. gonorrhoeae mRNA encoding a N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof; and(b) a nucleic acid sequence capable of controlling expression of the nucleic acid sequence encoding the mRNA encoding the N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, in a host cell, wherein the TbpB protein is an Ngo1 TbpB protein or an Ngo2 TbpB protein.

[0210] In another embodiment, the present disclosure provides, a host cell comprising a chimeric nucleic acid sequence comprising as operably linked components:(a) a nucleic acid sequence encoding an mRNA encoding a first and second N. gonorrhoeae TbpB protein, or immunogenically equivalent portions thereof,; and(b) one or more nucleic acid sequences capable of separately or jointly controlling expression of the nucleic acid sequence encoding the mRNA encoding the first and second N. gonorrhoeae TbpB protein, or immunogenically equivalent portions thereof, wherein the first and second TbpB protein are selected from an Ngo1 TbpB protein, and an Ngo2 TbpB protein, respectively.1007451850

[0211] In another embodiment, the present disclosure provide a mixture comprising a first and second host cell, the first host cell comprising a first chimeric nucleic acid comprising as operably linked components:(a) a nucleic acid sequence encoding an mRNA encoding a first N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof; and(b) a nucleic acid sequence capable of controlling expression of the nucleic acid sequence encoding the first mRNA encoding a first N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, in the first host cell, wherein the first TbpB protein is an Ngo1 TbpB protein; and the second host cell comprising a second chimeric nucleic acid comprising as operably linked components:(c) a nucleic acid sequence encoding an mRNA encoding a second N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof; and(d) a nucleic acid sequence capable of controlling expression of the nucleic acid sequence encoding the mRNA encoding the second N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, in the second host cell, wherein the second TbpB protein is an Ngo2 TbpB protein.

[0212] Turning next to the preparation of vaccine formulations, in an aspect hereof, in order to prepare a vaccine formulation, a preparation comprising a TbpB protein or immunogenically equivalent portion thereof, or mRNA encoding a TbpB protein, or immunogenically equivalent portion thereof, may be combined with at least one other pharmaceutically acceptable in ingredient, including, but not limited to, a diluent, an excipient, a carrier, an adjuvant, or mixtures thereof, whereby the TbpB protein or immunogenically equivalent portion thereof, or mRNA encoding a TbpB protein, or immunogenically equivalent portion thereof, and at least one other ingredient are mixed together or blended or homogenized or otherwise prepared until the vaccine formulation is formed.

[0213] The amount of TbpB protein or immunogenically equivalent portion thereof, or mRNA encoding a TbpB protein, or immunogenically equivalent portion1017451850thereof, in the vaccine formulation may vary. In general, consideration is given to the dose to be administered to a human. Doses for the TbpB protein or an immunogenically equivalent portion thereof, or mRNA encoding a TbpB protein, or immunogenically equivalent portion thereof, may be formulated to include TbpB protein or an immunogenically equivalent portion thereof, or mRNA encoding a TbpB protein, or immunogenically equivalent portion thereof, in quantities ranging from about 1 pg / kg of human body weight to about 0.25 mg / kg of human body weight, preferably about 1 pg / kg of human body weight to about 100 pg / kg of human body weight. Furthermore, vaccine formulations are preferably formulated so that a dose comprises at least about 0.001 % by weight or volume, at least 0.025% or about 0.025%, at least 0.05% or about 0.05%, at least 0.1 % or about 0.1 %, at least 0.5 or about 0.5%, at least 1 % or about 1 %, at least 5% or about 5%, at least 10% or about 10%, at least 15% or about 15%, at least 20% or about 20%, or at least 25% or about 25%, by weight of the TbpB protein, or mRNA encoding a TbpB protein, so that the ratio of TbpB, or mRNA encoding a TbpB protein, to other vaccine constituents (e.g., adjuvants, diluents, carriers, excipients) of the vaccine formulation by weight or volume is at least 0.001 :99.999, 0.025:99.975, 0.05: 99,95, 0.01 :99.99, 0.5:99.5, 1 :99, 5:95, 15:85, 20:80, or25:75, respectively, by weight or volume. The exact amount necessary, however, will vary depending on the age, and general condition of the recipient human to be treated, the severity of the condition being treated, the particular preparation delivered, the site of administration, as well as other factors. In this respect, vaccine formulations may, in particular, vary with respect to the quantity of TbpB protein, or immunogenically equivalent portion thereof, or mRNA encoding a TbpB protein, or immunogenically equivalent portion thereof, included in a dose. A suitable effective amount can be readily determined by one of skill in the art. Thus, a therapeutically effective amount of the TbpB protein or immunogenically equivalent portion, or mRNA encoding a TbpB protein, or immunogenically equivalent portion thereof, to be included in the vaccine formulations of the present disclosure will be an amount sufficient to bring about amelioration or prevention of disease or condition symptoms and will fall in a relatively broad range that can be determined through routine trials.

[0214] Vaccine formulations comprising the TbpB protein, or immunogenically equivalent portion thereof, or mRNA encoding a TbpB protein, or1027451850immunogenically equivalent portion thereof, of the present disclosure preferably further are prepared by combining the TbpB protein, or immunogenically equivalent portion thereof, or mRNA encoding a TbpB protein, or immunogenically equivalent portion thereof, with e.g., carriers, excipients, diluents, and auxiliary substances, such as wetting or emulsifying agents, pH buffering substances and the like. These carriers, excipients, diluents, and auxiliary substances are pharmaceutically acceptable ingredients. Pharmaceutically acceptable excipients include, but are not limited to, liquids such as water, saline, polyethyleneglycol, hyaluronic acid, glycerol, and ethanol. Pharmaceutically acceptable salts can also be included in the formulation, for example, mineral acid salts such as hydrochlorides, phosphates, sulfates, and the like; and the salts of organic acids such as acetates, propionates, benzoates, and the like. It is also preferred, although not required, that the vaccine formulation will contain a pharmaceutically acceptable carrier that serves as a stabilizer, particularly in order to stabilize the polypeptides of the present disclosure. Examples of suitable carriers that also act as stabilizers for peptides include, without limitation, pharmaceutical grades of dextrose, sucrose, lactose, sorbitol, inositol, dextran, and the like. Other suitable carriers include, again without limitation, starch, cellulose, sodium or calcium phosphates, citric acid, glycine, polyethylene glycols (PEGs), and combinations thereof. Carriers, excipients, diluents, may constitute, for example, from about 10% to about 95% by weight or volume of the vaccine formulation.

[0215] Further, auxiliary agents such as freeze-drying stabilizers, wetting or emulsifying agents, pH buffering agents, gelling or viscosity enhancing additives, and preservatives may also be included in the vaccine formulations of the present disclosure. Vaccine formulations generally comprise less than about 5% by weight of such auxiliary agents.

[0216] In order to augment an immune response, the vaccine formulations provided herein further preferably include one or more adjuvants, such as pharmacological agents, cytokines, or the like. Suitable adjuvants include any substance that enhances the immune response of the recipient human to the immunogenic TbpB protein, or immunogenically equivalent portion thereof, or mRNA encoding a TbpB protein, or immunogenically equivalent portion thereof, of the disclosure. Non-limiting examples of adjuvants include cytokines, e.g., IL-1 , IL- 2, IL-12, IL-6, and further include inorganic salts, e.g., aluminum hydroxide,1037451850aluminum phosphate, and calcium phosphate; oil emulsions, e.g., mineral oil, MF59, QS-21 , MontanideTmISA51 VG, or MontanideTmISA-720; Isocoms, e.g., ISCOMATRIX; microbial derivatives, e.g., monophosphoryl lipid A (MPLA), macrophage-activating protein-2, virosomes, LT / CT, CpG; natural polymers, e.g., polysaccharides; and synthetic polymers, e.g., polyanhydrides and polyesters, or nucleic acid analogs, such as Poly I :C. Adjuvants may be administered, for example, as proteins or other macromolecules at the same time (e.g., by inclusion in the vaccine formulation), prior to, or subsequent to, administration of the polypeptide antigens. When included in a vaccine formulation, adjuvants may constitute, for example, from 0.1 % or about 0.1 % to 50% or about 50%, from 0.1 % or about 0.1 % to 20% or about 20%, or from 1 % or about 1 % to 10% or about 10% by weight or volume of a vaccine formulation.

[0217] Furthermore, vaccine formulations comprising outer membrane vesicles (OMVs) comprising TbpB proteins may be prepared. In this respect, OMVs may be prepared from recombinant cells, for example E. coli cells, expressing TbpB proteins, as hereinbefore described. OMVs may be prepared from recombinant cells using techniques known to those of skill in the art, including, for example, as described by Thoma, J. and Burmann, B., 2020, in Bioprotocol 10(24): e3853 DOI :10.21769 / BioProtoc.3853. The OMVs comprising TbpB proteins may be used, togetherwith ingredients, such as excipients, diluents, carriers, and adjuvants, for example, as hereinbefore described, to prepare a vaccine formulation.

[0218] Thus, in another aspect, the present disclosure further provides, in at least one other embodiment, a method for preparing vaccine formulation comprising an effective amount of at least two N. gonorrhoeae TbpB proteins, or immunogenically equivalent portions thereof, or at least two mRNAs encoding two TbpB proteins, the vaccine formulation comprising at least one Ngo1 TbpB protein, and at least one Ngo2 TbpB protein, the method comprising:(a) providing a first chimeric nucleic acid sequence comprising as operably linked components:(i) a nucleic acid sequence encoding a first N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, or an mRNA encoding a first N. gonorrhoeae TbpB protein, or an1047451850immunogenically equivalent portion thereof, wherein the first TbpB protein is an Ngo1 protein; and(ii) one or more nucleic acid sequences capable of controlling expression of the nucleic acid sequence encoding the first TbpB protein, or immunogenically equivalent portion thereof, or the mRNA encoding the first N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, in a host cell;(b) providing a second chimeric nucleic acid sequence comprising as operably linked components:(i) a nucleic acid sequence encoding a second N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, or an mRNA encoding a second N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, wherein the second TbpB protein is an Ngo2 protein; and(ii) one or more nucleic acid sequences capable of controlling expression of the nucleic acid sequence encoding the second TbpB protein, or immunogenically equivalent portion thereof, or an mRNA encoding a first N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, in a host cell;(c) introducing the first and second chimeric nucleic acid sequence each into a separate first and second host cell, or together in a common host cell;(d) growing the first and second separate host cells, or common host cell, to produce the first and second TbpB proteins, or immunogenically equivalent portions thereof, or the mRNA encoding the first or second N. gonorrhoeae TbpB proteins, or an immunogenically equivalent portion thereof;(e) recovering the first and second TbpB proteins, or immunogenically equivalent portions thereof, or the mRNA encoding the first or second N. gonorrhoeae TbpB proteins, or an immunogenically equivalent portion thereof; and(f) formulating the first and second TbpB proteins, or immunogenically equivalent portions thereof, or the mRNA encoding the first and second N. gonorrhoeae TbpB proteins, or an immunogenically equivalent portion1057451850thereof, together with a pharmaceutically acceptable adjuvant to form a vaccine formulation comprising an effective amount of the first and second TbpB protein, or immunogenically equivalent portions thereof, to treat a human susceptible to i\i. gonorrhoeae infection.

[0219] The vaccine formulations of the present disclosure may be used to prevent infection or disease caused by pathogenic infectious i\i. gonorrhoeae in humans. The vaccine formulations may be used to immunize any human. Upon administration to a human clinical signs associated with N. gonorrhoeae infection may be diminished, such clinical signs include one or more of dysuria, pharyngitis, abnormal penile discharge, abnormal vaginal discharge, disseminated gonococcal infection (DGI), including DGI-associated arthritis, tenosynovitis, endocarditis and / or dermatitis. In general, such diminishing of clinical signs can be assessed relative to a human infected by N. gonorrhoeae not having been administered the vaccine formulation.

[0220] The vaccine formulations of the present may be administered to human using any convenient administration means. Thus, for example, the vaccine formulations may be injected, for example, intramuscularly or subcutaneously, or the vaccine formulations may be orally administered to the human. It will be understood that, in this respect, the administration means and techniques, such as, for example, in the case of injections, the gauge of the injection needle, may vary depending on for example, the age of the human. The dosage of the vaccine formulation will be dependent upon the disease, the route of administration, body weight, and other standard factors. In this respect, a person of ordinary skill in the art can readily titrate the appropriate dosage for an effective amount as well as select a suitable method of administration.

[0221] It is further noted that the vaccine formulations of the present disclosure may be administered prophylactically, i.e., in order to prevent N. gonorrhoeae infection in a human, or in order to ameliorate symptoms associated with N. gonorrhoeae infection following the occurrence of an infection in a human.

[0222] The administration of the vaccine formulations of the present disclosure generally elicits an immune response in the subject human. It will be understood that vaccine formulations comprising mRNAs encoding TbpB proteins may upon administration to a subject human elicit an in vivo production of the TbpB proteins in the subject human. Moreover, antibodies against a TbpB protein1067451850included in the vaccine formulation, or TbpB proteins produced in vivo may be formed by the human. In some embodiments, anti-TbpB antibodies can be detected in the blood serum of the human at least 7 days, at least 2 weeks, at least 5 weeks, at least 10 weeks, at least 13 weeks, at least 26 weeks, or at least 52 weeks following administration of the vaccine formulation.

[0223] In light of the foregoing, it will now be understood that, in another aspect, the present disclosure provides, in an example embodiment, a use of an Ngo1 TbpB protein, or an immunogenically equivalent portion thereof, together with an Ngo2 TbpB protein, or an immunogenically equivalent portion thereof, or a use of an mRNA encoding an Ngo1 TbpB protein, or an immunogenically equivalent portion thereof, together with an mRNA encoding an Ngo2 TbpB protein, or an immunogenically equivalent portion thereof, to prepare a vaccine formulation comprising the Ngo1 and Ngo2 TbpB proteins, or immunogenically equivalent portions thereof, or a vaccine formulation comprising the mRNAs encoding the Ngo1 and Ngo2 proteins, or immunologically equivalent portions thereof, together with a pharmaceutically acceptable adjuvant.

[0224] In light of the foregoing, it will now further be understood that, in another aspect, the present disclosure provides, in another example embodiment, a use of a vaccine formulation in the manufacture of a medicament for the prevention, treatment or amelioration of N. gonorrhoeae infection in a human in need thereof, the vaccine formulation comprising an effective amount of (i) at least two N. gonorrhoeae TbpB proteins, or immunogenically equivalent portions thereof, or (ii) at least two mRNA polynucleotides encoding the two N. gonorrhoeae TbpB proteins, or immunogenically equivalent portions thereof, the at least two N. gonorrhoeae TbpB proteins being selected from at least one Ngo1 TbpB protein, and at least one Ngo2 TbpB protein.

[0225] In light of the foregoing, it will now further be understood that, in another aspect, the present disclosure provides, in another example embodiment, a use of a vaccine formulation for the prevention or amelioration of N. gonorrhoeae infection in a human, the vaccine formulation comprising an effective amount of (i) at least two N. gonorrhoeae TbpB proteins, or immunogenically equivalent portions thereof, or (ii) at least two mRNA polynucleotides encoding the two N. gonorrhoeae TbpB proteins, or immunogenically equivalent portions thereof, the1077451850at least two / V. gonorrhoeae TbpB proteins being selected from at least one Ngo1 TbpB protein, and at least one Ngo2 TbpB protein.

[0226] As can now be understood, vaccine formulations comprising at least two TbpB proteins, or immunogenically equivalent portions thereof, or at least two mRNAs encoding TbpB proteins, or immunologically equivalent portions thereof, may be prepared. The vaccine formulations can be administered to a human to ameliorate or prevent infection of the human by N. gonorrhoeae.

[0227] Of course, the above-described example embodiments of the present disclosure are intended to be illustrative and in no way limiting. The embodiments are susceptible to many modifications or composition, details, and order of operation. The invention and this disclosure are intended to encompass all such modifications within its scope, as defined by the claims, which should be given a broad interpretation consistent with the description as a whole.SUMMARY OF SEQUENCES

[0228] SEQ.ID NO: 1 and SEQ.ID NO: 2 set forth the native polynucleotide sequence and deduced amino acid, respectively, of a TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the polypeptide also referred to as “27230”).

[0229] SEQ.ID NO: 3 and SEQ.ID NO: 4 set forth the polynucleotide sequence (E. coli codon optimized) and deduced amino acid, respectively, of a TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the polypeptide also referred to as “27230”).

[0230] SEQ.ID NO: 5 and SEQ.ID NO: 6 set forth the native polynucleotide sequence and deduced amino acid, respectively, of a TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the polypeptide also referred to as “27464”).

[0231] SEQ.ID NO: 7 and SEQ.ID NO: 8 set forth the polynucleotide sequence (E. coli codon optimized) and deduced amino acid, respectively, of a TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the polypeptide also referred to as “27464”).

[0232] SEQ.ID NO: 9 and SEQ.ID NO: 10 set forth the native polynucleotide sequence and deduced amino acid, respectively, of a TbpB of N. gonorrhoeae1087451850(belonging to phylogenetic cluster Ngo2, the polypeptide also referred to as “41385”).

[0233] SEQ.ID NO: 11 and SEQ.ID NO: 12 set forth the polynucleotide sequence (E. coli codon optimized) and deduced amino acid, respectively, of a TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the polypeptide also referred to as “41385”).

[0234] SEQ.ID NO: 13 and SEQ.ID NO: 14 set forth the native polynucleotide sequence and deduced amino acid, respectively, of a secreted TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the polypeptide also referred to as “41385”).

[0235] SEQ.ID NO: 15 and SEQ.ID NO: 16 set forth the native polynucleotide sequence and deduced amino acid, respectively, of a mutated (G222R) secreted TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “41385”).

[0236] SEQ.ID NO: 17 and SEQ.ID NO: 18 set forth the native polynucleotide sequence and deduced amino acid, respectively, of a mutated (E223R) secreted TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “41385”).

[0237] SEQ.ID NO: 19 and SEQ.ID NO: 20 set forth the native polynucleotide sequence and deduced amino acid, respectively, of a TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , also referred to as “48627”).

[0238] SEQ.ID NO: 21 and SEQ.ID NO: 22 set forth the polynucleotide sequence (E. coli codon optimized) and deduced amino acid, respectively, of a TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the polypeptide also referred to as “48627”).

[0239] SEQ.ID NO: 23 and SEQ.ID NO: 24 set forth the native polynucleotide sequence and deduced amino acid, respectively, of a secreted TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the polypeptide also referred to as “48627”).

[0240] SEQ.ID NO: 25 and SEQ.ID NO: 26 set forth the native polynucleotide sequence and deduced amino acid, respectively, of a mutated (D209H) secreted TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “48627”).1097451850

[0241] SEQ.ID NO: 27 and SEQ.ID NO: 28 set forth the native polynucleotide sequence and deduced amino acid, respectively, of a TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the polypeptide also referred to as “WHO Y”).

[0242] SEQ.ID NO: 29 and SEQ.ID NO: 30 set forth the polynucleotide sequence (E. coli codon optimized) and deduced amino acid, respectively, of a TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the polypeptide also referred to as “WHO Y”).

[0243] SEQ.ID NO: 31 , SEQ.ID NO: 32 and SEQ.ID NO: 33 set forth the native polynucleotide sequence, an E. coli optimized polynucleotide sequence, and deduced amino acid, respectively, of a TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the polypeptide also referred to as “32017”).

[0244] SEQ.ID NO: 34 and SEQ.ID NO: 35 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the polypeptide also referred to as “32017”).

[0245] SEQ.ID NO: 36, SEQ.ID NO: 37 and SEQ.ID NO: 38 set forth the native polynucleotide sequence, an E. coli optimized polynucleotide sequence, and deduced amino acid, respectively, of a TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the polypeptide also referred to as “32079”).

[0246] SEQ.ID NO: 39 and SEQ.ID NO: 40 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the polypeptide also referred to as “32079”).

[0247] SEQ.ID NO: 41 , SEQ.ID NO: 42 and SEQ.ID NO: 43 set forth the native polynucleotide sequence, an E. coli optimized polynucleotide sequence, and deduced amino acid, respectively, of a TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the polypeptide also referred to as “36247”).

[0248] SEQ.ID NO: 44 and SEQ.ID NO: 45 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the polypeptide also referred to as “36247”).

[0249] SEQ.ID NO: 46, SEQ.ID NO: 47 and SEQ.ID NO: 48 set forth the native polynucleotide sequence, an E. coli optimized polynucleotide sequence,1107451850and deduced amino acid, respectively, of a TbpB of / V. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the polypeptide also referred to as “47626”).

[0250] SEQ.ID NO: 49 and SEQ.ID NO: 50 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the polypeptide also referred to as “47626”).

[0251] SEQ.ID NO: 51 , SEQ.ID NO: 52 and SEQ.ID NO: 53 set forth the native polynucleotide sequence, an E. coli optimized polynucleotide sequence, and deduced amino acid, respectively, of a TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the polypeptide also referred to as “75977”).

[0252] SEQ.ID NO: 54 and SEQ.ID NO: 55 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the polypeptide also referred to as “75977”).

[0253] SEQ.ID NO: 56, SEQ.ID NO: 57 and SEQ.ID NO: 58 set forth the native polynucleotide sequence, an E. coli optimized polynucleotide sequence, and deduced amino acid, respectively, of a TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the polypeptide also referred to as “78646”).

[0254] SEQ.ID NO: 59 and SEQ.ID NO: 60 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the polypeptide also referred to as “78646”).

[0255] SEQ.ID NO: 61 and SEQ.ID NO: 62 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated K182A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27230”).

[0256] SEQ.ID NO: 63 and SEQ.ID NO: 64 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated K182E TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27230”).

[0257] SEQ.ID NO: 65 and SEQ.ID NO: 66 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated K192A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27230”).1117451850

[0258] SEQ.ID NO: 67 and SEQ.ID NO: 68 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated R196A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27230”).

[0259] SEQ.ID NO: 69 and SEQ.ID NO: 70 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated R196D TbpB of / V. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27230”).

[0260] SEQ.ID NO: 71 and SEQ.ID NO: 72 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D205ATbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27230”).

[0261] SEQ.ID NO: 73 and SEQ.ID NO: 74 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D205H TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27230”).

[0262] SEQ.ID NO: 75 and SEQ.ID NO: 76 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D205K TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27230”).

[0263] SEQ.ID NO: 77 and SEQ.ID NO: 78 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated R185A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27464”). 264] SEQ.ID NO: 79 and SEQ.ID NO: 80 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated R185D TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27464”).

[0265] SEQ.ID NO: 79 and SEQ.ID NO: 80 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated K185DTbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27464”).

[0266] SEQ.ID NO: 81 and SEQ.ID NO: 82 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated K195A1127451850TbpB of / V. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27464”).

[0267] SEQ.ID NO: 83 and SEQ.ID NO: 84 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated K195E TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27464”).

[0268] SEQ.ID NO: 85 and SEQ.ID NO: 86 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D208A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27464”).

[0269] SEQ.ID NO: 87 and SEQ.ID NO: 88 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D208R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27464”).

[0270] SEQ.ID NO: 89 and SEQ.ID NO: 90 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated K199A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “41385”).

[0271] SEQ.ID NO: 91 and SEQ.ID NO: 92 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated K199E TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “41385”).

[0272] SEQ.ID NO: 93 and SEQ.ID NO: 94 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated K208A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “41385”).

[0273] SEQ.ID NO: 95 and SEQ.ID NO: 96 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated R209A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “41385”).

[0274] SEQ.ID NO: 97 and SEQ.ID NO: 98 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated G222A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “41385”).1137451850

[0275] SEQ.ID NO: 99 and SEQ.ID NO: 100 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated E223A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “41385”).

[0276] SEQ.ID NO: 101 and SEQ.ID NO: 102 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated K186A TbpB of / V. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “48627”).

[0277] SEQ.ID NO: 103 and SEQ.ID NO: 104 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated K186E TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “48627”).

[0278] SEQ.ID NO: 105 and SEQ.ID NO: 106 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated K196A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “48627”).

[0279] SEQ.ID NO: 107 and SEQ.ID NO: 108 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated R200A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “48627”).

[0280] SEQ.ID NO: 109 and SEQ.ID NO: 110 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated R200D TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “48627”).

[0281] SEQ.ID NO: 111 and SEQ.ID NO: 112 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D209A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “48627”).

[0282] SEQ.ID NO: 113 and SEQ.ID NO: 114 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated K188A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “WHO Y”).

[0283] SEQ.ID NO: 115 and SEQ.ID NO: 116 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated K188E1147451850TbpB of / V. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “WHO Y”).

[0284] SEQ.ID NO: 117 and SEQ.ID NO: 118 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated K198A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “WHO Y”).

[0285] SEQ.ID NO: 119 and SEQ.ID NO: 120 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated R202A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “WHO Y”).

[0286] SEQ.ID NO: 121 and SEQ.ID NO: 122 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated R202D TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “WHO Y”).

[0287] SEQ.ID NO: 123 and SEQ.ID NO: 124 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D225A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “WHO Y”).

[0288] SEQ.ID NO: 125 and SEQ.ID NO: 126 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D225H TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “WHO Y”).

[0289] SEQ.ID NO: 127 and SEQ.ID NO: 128 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D225K TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “WHO Y”).

[0290] SEQ.ID NO: 129 is polynucleotide primer sequence referred to as “F- RF”.

[0291] SEQ.ID NO: 130 is polynucleotide primer sequence referred to as “R-RF”.

[0292] SEQ.ID NO: 131 is polynucleotide primer sequence referred to as “ngo-C2FORx2”.

[0293] SEQ.ID NO: 132 is polynucleotide primer sequence referred to as “ngo-C2REVx”.1157451850

[0294] SEQ.ID NO: 133 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated R185A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27464”).

[0295] SEQ.ID NO: 134 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated R185A TbpB of / V. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27464”).

[0296] SEQ.ID NO: 135 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated R185D TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27464”).

[0297] SEQ.ID NO: 136 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated R185D TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27464”).

[0298] SEQ.ID NO: 137 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated K195A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27464”).

[0299] SEQ.ID NO: 138 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated K195A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27464”).

[0300] SEQ.ID NO: 139 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated K195E TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27464”).

[0301] SEQ.ID NO: 140 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated K195E TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27464”).

[0302] SEQ.ID NO: 141 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated D208A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27464”).

[0303] SEQ.ID NO: 142 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated D208A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27464”).

[0304] SEQ.ID NO: 143 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated D208R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27464”).1167451850

[0305] SEQ.ID NO: 144 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated D208R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27464”).

[0306] SEQ.ID NO: 145 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated K182A TbpB of / V. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27230”).

[0307] SEQ.ID NO: 146 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated K182A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27230”).

[0308] SEQ.ID NO: 147 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated K182E TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27230”).

[0309] SEQ.ID NO: 148 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated K182E TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27230”).

[0310] SEQ.ID NO: 149 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated K192A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27230”).

[0311] SEQ.ID NO: 150 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated K192A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27230”).

[0312] SEQ.ID NO: 151 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated R196A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27230”).

[0313] SEQ.ID NO: 152 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated R196A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27230”).

[0314] SEQ.ID NO: 153 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated R196E TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27230”).

[0315] SEQ.ID NO: 154 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated R196E TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27230”).1177451850

[0316] SEQ.ID NO: 155 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated D205A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27230”).

[0317] SEQ.ID NO: 156 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated D205A TbpB of / V. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27230”).

[0318] SEQ.ID NO: 157 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated D205H TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27230”).

[0319] SEQ.ID NO: 158 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated D205H TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27230”).

[0320] SEQ.ID NO: 159 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated D205E TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27230”).

[0321] SEQ.ID NO: 160 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated D205E TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “27230”).

[0322] SEQ.ID NO: 161 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated K199A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “41385”).

[0323] SEQ.ID NO: 162 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated K199A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “41385”).

[0324] SEQ.ID NO: 163 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated K199E TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “41385”).

[0325] SEQ.ID NO: 164 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated K199E TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “41385”).

[0326] SEQ.ID NO: 165 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated K208A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “41385”).1187451850

[0327] SEQ.ID NO: 166 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated K208A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “41385”).

[0328] SEQ.ID NO: 167 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated R209A TbpB of / V. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “41385”).

[0329] SEQ.ID NO: 168 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated R209A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “41385”).

[0330] SEQ.ID NO: 169 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated G222A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “41385”).

[0331] SEQ.ID NO: 170 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated G222A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “41385”).

[0332] SEQ.ID NO: 171 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated G222R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “41385”).

[0333] SEQ.ID NO: 172 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated G222R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “41385”).

[0334] SEQ.ID NO: 173 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated E223A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “41385”).

[0335] SEQ.ID NO: 174 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated E223A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “41385”).

[0336] SEQ.ID NO: 175 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated E223R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “41385”).

[0337] SEQ.ID NO: 176 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated E223R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “41385”).1197451850

[0338] SEQ.ID NO: 177 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated K186A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “48627”).

[0339] SEQ.ID NO: 178 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated K186A TbpB of / V. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “48627”).

[0340] SEQ.ID NO: 179 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated K186E TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “48627”).

[0341] SEQ.ID NO: 180 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated K186E TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “48627”).

[0342] SEQ.ID NO: 181 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated K196A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “48627”).

[0343] SEQ.ID NO: 182 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated K196A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “48627”).

[0344] SEQ.ID NO: 183 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated R200A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “48627”).

[0345] SEQ.ID NO: 184 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated R200A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “48627”).

[0346] SEQ.ID NO: 185 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated R200D TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “48627”).

[0347] SEQ.ID NO: 186 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated R200D TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “48627”).

[0348] SEQ.ID NO: 187 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated D209A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “48627”).1207451850

[0349] SEQ.ID NO: 188 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated D209A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “48627”).

[0350] SEQ.ID NO: 189 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated D209H TbpB of / V. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “48627”).

[0351] SEQ.ID NO: 190 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated D209H TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “48627”).

[0352] SEQ.ID NO: 191 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated K188A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “WHO Y”).

[0353] SEQ.ID NO: 192 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated K188A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “WHO Y”).

[0354] SEQ.ID NO: 193 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated K188E TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “WHO Y”).

[0355] SEQ.ID NO: 194 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated K188E TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “WHO Y”).

[0356] SEQ.ID NO: 195 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated K198A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “WHO Y”).

[0357] SEQ.ID NO: 196 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated K198A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “WHO Y”).

[0358] SEQ.ID NO: 197 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated R202A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “WHO Y”).

[0359] SEQ.ID NO: 198 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated R202A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “WHO Y”).1217451850

[0360] SEQ.ID NO: 199 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated R202D TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “WHO Y”).

[0361] SEQ.ID NO: 200 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated R202D TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “WHO Y”).

[0362] SEQ.ID NO: 201 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated D225A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “WHO Y”).

[0363] SEQ.ID NO: 202 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated D225A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “WHO Y”).

[0364] SEQ.ID NO: 203 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated D225H TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “WHO Y”).

[0365] SEQ.ID NO: 204 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated D225H TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “WHO Y”).

[0366] SEQ.ID NO: 205 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated D225K TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “WHO Y”).

[0367] SEQ.ID NO: 206 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated D225K TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo1 , the native polypeptide also referred to as “WHO Y”).

[0368] SEQ.ID NO: 207 and SEQ.ID NO: 208 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D202A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “32017”).

[0369] SEQ.ID NO: 209 and SEQ.ID NO: 210 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D202R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “32017”).

[0370] SEQ.ID NO: 211 and SEQ.ID NO: 212 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated E221A1227451850TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “32017”).

[0371] SEQ.ID NO: 213 and SEQ.ID NO: 214 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated E221 R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “32017”).

[0372] SEQ.ID NO: 215 and SEQ.ID NO: 216 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D203A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “32079”).

[0373] SEQ.ID NO: 217 and SEQ.ID NO: 218 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D203R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “32079”).

[0374] SEQ.ID NO: 219 and SEQ.ID NO: 220 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D221 K,D223K,D225K TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “32079”).

[0375] SEQ.ID NO: 221 and SEQ.ID NO: 222 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D223K TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “32079”).

[0376] SEQ.ID NO: 223 and SEQ.ID NO: 224 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D230A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “32079”).

[0377] SEQ.ID NO: 225 and SEQ.ID NO: 226 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D230R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “32079”).

[0378] SEQ.ID NO: 227 and SEQ.ID NO: 228 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated E205A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “36247”).1237451850

[0379] SEQ.ID NO: 229 and SEQ.ID NO: 230 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated E205K TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “36247”).

[0380] SEQ.ID NO: 231 and SEQ.ID NO: 232 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated E221A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “36247”).

[0381] SEQ.ID NO: 233 and SEQ.ID NO: 234 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated E221 K TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “36247”).

[0382] SEQ.ID NO: 235 and SEQ.ID NO: 236 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D202A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “47626”).

[0383] SEQ.ID NO: 237 and SEQ.ID NO: 238 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D202K TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “47626”).

[0384] SEQ.ID NO: 239 and SEQ.ID NO: 240 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated E205A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “47626”).

[0385] SEQ.ID NO: 241 and SEQ.ID NO: 242 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated E205R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “47626”).

[0386] SEQ.ID NO: 243 and SEQ.ID NO: 244 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D221A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “47626”).

[0387] SEQ.ID NO: 245 and SEQ.ID NO: 246 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D221 K1247451850TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “47626”).

[0388] SEQ.ID NO: 247 and SEQ.ID NO: 248 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D119A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “75977”).

[0389] SEQ.ID NO: 249 and SEQ.ID NO: 250 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D119R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “75977”).

[0390] SEQ.ID NO: 251 and SEQ.ID NO: 252 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D202A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “75977”).

[0391] SEQ.ID NO: 253 and SEQ.ID NO: 254 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D202R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “75977”).

[0392] SEQ.ID NO: 255 and SEQ.ID NO: 256 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D220A.D222A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “75977”).

[0393] SEQ.ID NO: 257 and SEQ.ID NO: 258 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D220R.D222R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “75977”).

[0394] SEQ.ID NO: 259 and SEQ.ID NO: 260 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D202A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “78646”).

[0395] SEQ.ID NO: 261 and SEQ.ID NO: 262 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D202R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “78646”).1257451850

[0396] SEQ.ID NO: 263 and SEQ.ID NO: 264 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated E204A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “78646”).

[0397] SEQ.ID NO: 265 and SEQ.ID NO: 266 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated E204K TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “78646”).

[0398] SEQ.ID NO: 267 and SEQ.ID NO: 268 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D221A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “78646”).

[0399] SEQ.ID NO: 269 and SEQ.ID NO: 270 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D221 R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “78646”).

[0400] SEQ.ID NO: 271 and SEQ.ID NO: 272 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D267A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “78646”).

[0401] SEQ.ID NO: 273 and SEQ.ID NO: 274 set forth the polynucleotide sequence and deduced amino acid, respectively, of a secreted mutated D267R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “78646”).

[0402] SEQ.ID NO: 275 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated D202A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “32017”).

[0403] SEQ.ID NO: 276 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated D202A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “32017”).

[0404] SEQ.ID NO: 277 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated D202R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “32017”).1267451850

[0405] SEQ.ID NO: 278 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated D202R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “32017”).

[0406] SEQ.ID NO: 279 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated E221A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “32017”).

[0407] SEQ.ID NO: 280 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated E221A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “32017”).

[0408] SEQ.ID NO: 281 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated E221 R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “32017”).

[0409] SEQ.ID NO: 282 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated E221 R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “32017”).

[0410] SEQ.ID NO: 283 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated D203A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “32079”).

[0411] SEQ.ID NO: 284 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated D203A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “32079”).

[0412] SEQ.ID NO: 285 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated D203R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “32079”).

[0413] SEQ.ID NO: 286 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated D203R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “32079”).

[0414]

[0415] SEQ.ID NO: 287 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated D221 K, D223K, D225K TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “32079”).

[0416] SEQ.ID NO: 288 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated D221 K, D223K, D225K TbpB of N. gonorrhoeae1277451850(belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “32079”).

[0417] SEQ.ID NO: 289 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated D223K TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “32079”).

[0418] SEQ.ID NO: 290 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated D223K TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “32079”).

[0419] SEQ.ID NO: 291 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated D230A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “32079”).

[0420] SEQ.ID NO: 292 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated D230A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “32079”).

[0421] SEQ.ID NO: 293 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated D230R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “32079”).

[0422] SEQ.ID NO: 294 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated D230R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “32079”).

[0423] SEQ.ID NO: 295 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated E208A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “36247”).

[0424] SEQ.ID NO: 296 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated E208A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “36247”).

[0425] SEQ.ID NO: 297 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated E205K TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “36247”).

[0426] SEQ.ID NO: 298 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated E205K TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “36247”).1287451850

[0427] SEQ.ID NO: 299 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated E221A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “36247”).

[0428] SEQ.ID NO: 300 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated E221A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “36247”).

[0429] SEQ.ID NO: 301 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated E221 K TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “36247”).

[0430] SEQ.ID NO: 302 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated E221 K TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “36247”).

[0431] SEQ.ID NO: 303 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated D202A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “47626”).

[0432] SEQ.ID NO: 304 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated D202A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “47626”).

[0433] SEQ.ID NO: 305 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated D202K TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “47626”).

[0434] SEQ.ID NO: 306 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated D202K TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “47626”).

[0435] SEQ.ID NO: 307 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated E205A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “47626”).

[0436] SEQ.ID NO: 308 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated E205A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “47626”).

[0437] SEQ.ID NO: 309 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated E205R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “47626”).1297451850

[0438] SEQ.ID NO: 310 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated E205R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “47626”).

[0439] SEQ.ID NO: 311 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated D221A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “47626”).

[0440] SEQ.ID NO: 312 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated D221A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “47626”)

[0441] SEQ.ID NO: 313 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated D221 K TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “47626”).

[0442] SEQ.ID NO: 314 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated D221 K TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “47626”).

[0443] SEQ.ID NO: 315 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated D119A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “75977”).

[0444] SEQ.ID NO: 316 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated D119A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “75977”)

[0445] SEQ.ID NO: 317 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated D119R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “75977”).

[0446] SEQ.ID NO: 318 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated D119R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “75977”).

[0447] SEQ.ID NO: 319 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated D202A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “75977”).

[0448] SEQ.ID NO: 320 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated D202A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “75977”).1307451850

[0449] SEQ.ID NO: 321 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated D202R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “75977”).

[0450] SEQ.ID NO: 322 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated D202R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “75977”).

[0451] SEQ.ID NO: 323 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated D220A.D222A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “75977”).

[0452] SEQ.ID NO: 324 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated D220A.D222A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “75977”)

[0453] SEQ.ID NO: 325 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated D220R.D222R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “75977”).

[0454] SEQ.ID NO: 326 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated D220R.D222R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “75977”).

[0455] SEQ.ID NO: 327 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated D202A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “78646”).

[0456] SEQ.ID NO: 328 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated D202A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “78646”)

[0457] SEQ.ID NO: 329 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated D202R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “78646”).

[0458] SEQ.ID NO: 330 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated D202R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “78646”).1317451850

[0459] SEQ.ID NO: 331 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated E204A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “78646”)

[0460] SEQ.ID NO: 332 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated E204A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “78646”).

[0461] SEQ.ID NO: 333 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated E204K TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “78646”).

[0462] SEQ.ID NO: 334 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated E204K TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “78646”).

[0463] SEQ.ID NO: 335 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated E221A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “78646”).

[0464] SEQ.ID NO: 336 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated E221A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “78646”).

[0465] SEQ.ID NO: 337 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated E221 R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “78646”).

[0466] SEQ.ID NO: 338 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated E221 R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “78646”).

[0467] SEQ.ID NO: 339 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated D267A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “78646”).

[0468] SEQ.ID NO: 340 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated D267A TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “78646”).

[0469] SEQ.ID NO: 341 is polynucleotide primer sequence (forward) for use in preparing a secreted mutated D267R TbpB of N. gonorrhoeae (belonging to phylogenetic cluster Ngo2, the native polypeptide also referred to as “78646”).1327451850

[0470] SEQ.ID NO: 342 is polynucleotide primer sequence (reverse) for use in preparing a secreted mutated D267...

Claims

CLAIMS1. A vaccine formulation for the prevention or amelioration of / V. gonorrhoeae infection in a human, the vaccine formulation comprising an effective amount of (i) at least two N. gonorrhoeae TbpB proteins, or immunogenically equivalent portions thereof, or (ii) at least two mRNA polynucleotides encoding the two N. gonorrhoeae TbpB proteins, or immunogenically equivalent portions thereof, the at least two N. gonorrhoeae TbpB proteins being selected from at least one Ngo1 TbpB protein, and at least one Ngo2 TbpB protein.

2. A vaccine formulation according to claim 1 , wherein the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, is a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 1 , SEQ.ID NO: 3, SEQ.ID NO: 5, SEQ.ID NO: 7, SEQ.ID NO: 19, SEQ.ID NO: 21 , SEQ.ID NO: 23, SEQ.ID NO: 25, SEQ.ID NO: 27, SEQ.ID NO: 29, SEQ.ID NO: 61 , SEQ.ID NO: 63, SEQ.ID NO: 65, SEQ.ID NO: 67, SEQ.ID NO: 69, SEQ.ID NO: 71 , SEQ.ID NO: 73, SEQ.ID NO: 75, SEQ.ID NO: 77, SEQ.ID NO: 79, SEQ.ID NO: 81 , SEQ.ID NO: 83, SEQ.ID NO: 85, SEQ.ID NO: 87, SEQ.ID NO: 101 , SEQ.ID NO: 103, SEQ.ID NO: 105, SEQ.ID NO: 107, SEQ.ID NO: 109, SEQ.ID NO: 111 , SEQ.ID NO: 113, SEQ.ID NO: 115, SEQ.ID NO: 117, SEQ.ID NO: 119, SEQ.ID NO: 121 , SEQ.ID NO: 123, SEQ.ID NO: 125, or SEQ.ID NO: 127;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 2, SEQ.ID NO: 4, SEQ.ID NO: 6, SEQ.ID. NO: 8, SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24,1567451850SEQ.ID NO: 26, SEQ.ID NO: 28, SEQ.ID NO: 30, SEQ.ID NO: 62, SEQ.ID NO: 64, SEQ.ID NO: 66, SEQ.ID NO: 68, SEQ.ID NO: 70, SEQ.ID NO: 72, SEQ.ID NO: 74, SEQ.ID NO: 76, SEQ.ID NO: 78, SEQ.ID NO: 80, SEQ.ID NO: 82, SEQ.ID NO: 84, SEQ.ID NO: 86, SEQ.ID NO: 88, SEQ.ID NO: 102, SEQ.ID NO: 104, SEQ.ID NO: 106, SEQ.ID NO: 108, SEQ.ID NO: 110, SEQ.ID NO: 112 SEQ.ID NO: 114, SEQ.ID NO: 116, SEQ.ID NO: 187, SEQ.ID NO: 120, SEQ.ID NO: 122, SEQ.ID NO: 124, SEQ.ID NO: 126, or SEQ.ID NO: 128, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 2, SEQ.ID NO: 4, SEQ.ID NO: 6, SEQ.ID. NO: 8, SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, SEQ.ID NO: 26, SEQ.ID NO: 28, SEQ.ID NO: 30, SEQ.ID NO: 62, SEQ.ID NO: 64, SEQ.ID NO: 66, SEQ.ID NO: 68, SEQ.ID NO: 70, SEQ.ID NO: 72, SEQ.ID NO: 74, SEQ.ID NO: 76, SEQ.ID NO: 78, SEQ.ID NO: 80, SEQ.ID NO: 82, SEQ.ID NO: 84, SEQ.ID NO: 86, SEQ.ID NO: 88, SEQ.ID NO: 102, SEQ.ID NO: 104, SEQ.ID NO: 106, SEQ.ID NO: 108, SEQ.ID NO: 110, SEQ.ID NO: 112 SEQ.ID NO: 114, SEQ.ID NO: 116, SEQ.ID NO: 187, SEQ.ID NO: 120, SEQ.ID NO: 122, SEQ.ID NO: 124, SEQ.ID NO: 126, or SEQ.ID NO: 128, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

3. A vaccine formulation according to claim 1 , the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, is a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 25;(b) a nucleic acid sequence having at least 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with the nucleic acid sequence of (a);(c) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to the nucleic acid sequence of (a);1577451850(e) a nucleic acid sequence encoding a polypeptide having the amino acid sequences set forth in SEQ.ID NO: 26, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of the amino acid sequence set forth in SEQ.ID NO: 26, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

4. A vaccine formulation according to claim 1 , wherein the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, is a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 9, SEQ.ID NO: 11 , SEQ.ID NO: 13, SEQ.ID NO: 15, SEQ.ID NO: 17, SEQ.ID NO: 31 , SEQ.ID NO: 32, SEQ.ID NO: 34, SEQ.ID NO: 36, SEQ.ID NO: 37, SEQ.ID NO: 39, SEQ.ID NO: 41 , SEQ.ID NO: 42, SEQ.ID NO: 44, SEQ.ID NO: 46, SEQ.ID NO: 47, SEQ.ID NO: 49, SEQ.ID NO: 51 , SEQ.ID NO: 52, SEQ.ID NO: 54, SEQ.ID NO: 56, SEQ.ID NO: 57, SEQ.IDNO: 59, SEQ.ID NO: 89, SEQ.ID NO: 91 , SEQ.ID NO: 93, SEQ.ID NO: 95,SEQ.ID NO: 97, SEQ.ID NO: 99, SEQ.ID NO: 207, SEQ.ID NO: 209, SEQ.IDNO: 211 , SEQ.ID NO: 213, SEQ.ID NO: 215, SEQ.ID NO: 217, SEQ.ID NO:219, SEQ.ID NO: 221 , SEQ.ID NO: 223, SEQ.ID NO: 225, SEQ.ID NO: 227,SEQ.ID NO: 229, SEQ.ID NO: 231 , SEQ.ID NO: 233, SEQ.ID NO: 235SEQ.ID NO: 237, SEQ.ID NO: 239, SEQ.ID NO: 241 , SEQ.ID NO: 243SEQ.ID NO: 245, SEQ.ID NO: 247, SEQ.ID NO: 249, SEQ.ID NO: 251SEQ.ID NO: 253, SEQ.ID NO: 255, SEQ.ID NO: 257, SEQ.ID NO: 259SEQ.ID NO: 261 , SEQ.ID NO: 263, SEQ.ID NO: 265, SEQ.ID NO: 267SEQ.ID NO: 269, SEQ.ID NO: 271 , SEQ.ID NO: 273, SEQ.ID NO: 343, orSEQ.ID NO: 345;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;1587451850(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.ID NO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO: 216, SEQ.ID NO:218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224, SEQ.ID NO: 226,SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232, SEQ.ID NO: 234,SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240, SEQ.ID NO: 242,SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248, SEQ.ID NO: 250,SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256, SEQ.ID NO: 258,SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264, SEQ.ID NO: 266,SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272, SEQ.ID NO: 274, SEQ.ID NO: 344, or SEQ.ID NO: 3< 6, or an immunoc enically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.IDNO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.IDNO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO: 216, SEQ.ID NO218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224, SEQ.ID NO: 226SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232, SEQ.ID NO: 234,SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240, SEQ.ID NO: 242,SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248, SEQ.ID NO: 250,SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256, SEQ.ID NO: 258,SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264, SEQ.ID NO: 266,SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272, SEQ.ID NO: 274,1597451850SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

5. A vaccine formulation according to claim 1 , wherein the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, is a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 1 , SEQ.ID NO: 3, SEQ.ID NO: 5, SEQ.ID NO: 7, SEQ.ID NO: 19, SEQ.ID NO: 21 , SEQ.ID NO: 23, SEQ.ID NO: 25, SEQ.ID NO: 27, SEQ.ID NO: 29, SEQ.ID NO: 61 , SEQ.ID NO: 63, SEQ.ID NO: 65, SEQ.ID NO: 67, SEQ.ID NO: 69, SEQ.ID NO: 71 , SEQ.ID NO: 73, SEQ.ID NO: 75, SEQ.ID NO: 77, SEQ.ID NO: 79, SEQ.ID NO: 81 , SEQ.ID NO: 83, SEQ.ID NO: 85, SEQ.ID NO: 87, SEQ.ID NO: 101 , SEQ.ID NO: 103, SEQ.ID NO: 105, SEQ.ID NO: 107, SEQ.ID NO: 109, SEQ.ID NO: 111 , SEQ.ID NO: 113, SEQ.ID NO: 115, SEQ.ID NO: 117, SEQ.ID NO: 119, SEQ.ID NO: 121 , SEQ.ID NO: 123, SEQ.ID NO: 125, or SEQ.ID NO: 127;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 2, SEQ.ID NO: 4, SEQ.ID NO: 6, SEQ.ID. NO: 8, SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, SEQ.ID NO: 26, SEQ.ID NO: 28, SEQ.ID NO: 30, SEQ.ID NO: 62, SEQ.ID NO: 64, SEQ.ID NO: 66, SEQ.ID NO: 68, SEQ.ID NO: 70, SEQ.ID NO: 72, SEQ.ID NO: 74, SEQ.ID NO: 76, SEQ.ID NO: 78, SEQ.ID NO: 80, SEQ.ID NO: 82, SEQ.ID NO: 84, SEQ.ID NO: 86, SEQ.ID NO: 88, SEQ.ID NO: 102, SEQ.ID NO: 104, SEQ.ID NO: 106, SEQ.ID NO: 108, SEQ.ID NO: 110, SEQ.ID NO: 112 SEQ.ID NO: 114, SEQ.ID NO: 116, SEQ.ID NO: 187,1607451850SEQ.ID NO: 120, SEQ.ID NO: 122, SEQ.ID NO: 124, SEQ.ID NO: 126, or SEQ.ID NO: 128, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 2, SEQ.ID NO: 4, SEQ.ID NO: 6, SEQ.ID. NO: 8, SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, SEQ.ID NO: 26, SEQ.ID NO: 28, SEQ.ID NO: 30, SEQ.ID NO: 62, SEQ.ID NO: 64, SEQ.ID NO: 66, SEQ.ID NO: 68, SEQ.ID NO: 70, SEQ.ID NO: 72, SEQ.ID NO: 74, SEQ.ID NO: 76, SEQ.ID NO: 78, SEQ.ID NO: 80, SEQ.ID NO: 82, SEQ.ID NO: 84, SEQ.ID NO: 86, SEQ.ID NO: 88, SEQ.ID NO: 102, SEQ.ID NO: 104, SEQ.ID NO: 106, SEQ.ID NO: 108, SEQ.ID NO: 110, SEQ.ID NO: 112 SEQ.ID NO: 114, SEQ.ID NO: 116, SEQ.ID NO: 187, SEQ.ID NO: 120, SEQ.ID NO: 122, SEQ.ID NO: 124, SEQ.ID NO: 126, or SEQ.ID NO: 128, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f); and the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, is a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 9, SEQ.ID NO: 11 , SEQ.ID NO: 13, SEQ.ID NO: 15, SEQ.ID NO: 17, SEQ.ID NO: 31 , SEQ.ID NO: 32, SEQ.ID NO: 34, SEQ.ID NO: 36, SEQ.ID NO: 37, SEQ.ID NO: 39, SEQ.ID NO: 41 , SEQ.ID NO: 42, SEQ.ID NO: 44, SEQ.ID NO: 46, SEQ.ID NO: 47, SEQ.ID NO: 49, SEQ.ID NO: 51 , SEQ.ID NO: 52, SEQ.ID NO: 54, SEQ.ID NO: 56, SEQ.ID NO: 57, SEQ.ID NO: 59, SEQ.ID NO: 89, SEQ.ID NO: 91 , SEQ.ID NO: 93, SEQ.ID NO: 95, SEQ.ID NO: 97, SEQ.ID NO: 99, SEQ.ID NO: 207, SEQ.ID NO: 209, SEQ.IDNO: 211 , SEQ.ID NO: 213, SEQ.ID NO: 215, SEQ.ID NO: 217, SEQ.ID NO219, SEQ.ID NO: 221 , SEQ.ID NO: 223, SEQ.ID NO: 225, SEQ.ID NO: 227SEQ.ID NO: 229, SEQ.ID NO: 231 , SEQ.ID NO: 233, SEQ.ID NO: 235,SEQ.ID NO: 237, SEQ.ID NO: 239, SEQ.ID NO: 241 , SEQ.ID NO: 243,SEQ.ID NO: 245, SEQ.ID NO: 247, SEQ.ID NO: 249, SEQ.ID NO: 251 ,SEQ.ID NO: 253, SEQ.ID NO: 255, SEQ.ID NO: 257, SEQ.ID NO: 259,SEQ.ID NO: 261 , SEQ.ID NO: 263, SEQ.ID NO: 265, SEQ.ID NO: 267,1617451850SEQ.ID NO: 269, SEQ.ID NO: 271 , SEQ.ID NO: 273, SEQ.ID NO: 343, or SEQ.ID NO: 345;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.ID NO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO: 216, SEQ.ID NO: 218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224, SEQ.ID NO: 226,SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232, SEQ.ID NO: 234SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240, SEQ.ID NO: 242SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248, SEQ.ID NO: 250SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256, SEQ.ID NO: 258SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264, SEQ.ID NO: 266SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272, SEQ.ID NO: 274SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.ID NO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO: 216, SEQ.ID NO:1627451850218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224, SEQ.ID NO: 226,SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232, SEQ.ID NO: 234,SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240, SEQ.ID NO: 242,SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248, SEQ.ID NO: 250,SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256, SEQ.ID NO: 258,SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264, SEQ.ID NO: 266,SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272, SEQ.ID NO: 274,SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

6. A vaccine formulation according to claim 1 , wherein the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, is a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 19, SEQ.ID NO: 21 , SEQ.ID NO: 23, orSEQ.ID NO: 25;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, orSEQ.ID NO: 26, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, orSEQ.ID NO: 26, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f);1637451850and the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, is a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 9, SEQ.ID NO: 11 , SEQ.ID NO: 13, SEQ.ID NO: 15, SEQ.ID NO: 17, SEQ.ID NO: 31 , SEQ.ID NO: 32, SEQ.ID NO: 34, SEQ.ID NO: 36, SEQ.ID NO: 37, SEQ.ID NO: 39, SEQ.ID NO: 41 , SEQ.ID NO: 42, SEQ.ID NO: 44, SEQ.ID NO: 46, SEQ.ID NO: 47, SEQ.ID NO: 49, SEQ.ID NO: 51 , SEQ.ID NO: 52, SEQ.ID NO: 54, SEQ.ID NO: 56, SEQ.ID NO: 57, SEQ.ID NO: 59, SEQ.ID NO: 89, SEQ.ID NO: 91 , SEQ.ID NO: 93, SEQ.ID NO: 95,SEQ.ID NO: 97, SEQ.ID NO: 99, SEQ.ID NO: 207, SEQ.ID NO: 209, SEQ.IDNO: 211 , SEQ.ID NO: 213, SEQ.ID NO: 215, SEQ.ID NO: 217, SEQ.ID NO:219, SEQ.ID NO: 221 , SEQ.ID NO: 223, SEQ.ID NO: 225, SEQ.ID NO: 227,SEQ.ID NO: 229, SEQ.ID NO: 231 , SEQ.ID NO: 233, SEQ.ID NO: 235SEQ.ID NO: 237, SEQ.ID NO: 239, SEQ.ID NO: 241 , SEQ.ID NO: 243SEQ.ID NO: 245, SEQ.ID NO: 247, SEQ.ID NO: 249, SEQ.ID NO: 251SEQ.ID NO: 253, SEQ.ID NO: 255, SEQ.ID NO: 257, SEQ.ID NO: 259SEQ.ID NO: 261 , SEQ.ID NO: 263, SEQ.ID NO: 265, SEQ.ID NO: 267SEQ.ID NO: 269, SEQ.ID NO: 271 , SEQ.ID NO: 273, SEQ.ID NO: 343, or SEQ.ID NO: 345;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.ID NO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.ID1647451850NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO: 216, SEQ.ID NO: 218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224, SEQ.ID NO: 226,SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232, SEQ.ID NO: 234,SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240, SEQ.ID NO: 242,SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248, SEQ.ID NO: 250,SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256, SEQ.ID NO: 258,SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264, SEQ.ID NO: 266,SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272, SEQ.ID NO: 274,SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.ID NO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO: 216, SEQ.ID NO:218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224, SEQ.ID NO: 226,SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232, SEQ.ID NO: 234,SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240, SEQ.ID NO: 242,SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248, SEQ.ID NO: 250,SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256, SEQ.ID NO: 258,SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264, SEQ.ID NO: 266,SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272, SEQ.ID NO: 274,SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

7. A vaccine formulation according to claim 1 , wherein the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, is a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:1657451850(a) SEQ.ID NO: 25;(b) a nucleic acid sequence having at least 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with the nucleic acid sequence of (a);(c) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to the nucleic acid sequence of (a);(e) a nucleic acid sequence encoding a polypeptide having the amino acid sequences set forth in SEQ.ID NO: 26, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of the amino acid sequence set forth in SEQ.ID NO: 26, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f); and the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, is a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 9, SEQ.ID NO: 11 , SEQ.ID NO: 13, SEQ.ID NO: 15, SEQ.ID NO: 17, SEQ.ID NO: 31 , SEQ.ID NO: 32, SEQ.ID NO: 34, SEQ.ID NO: 36, SEQ.ID NO: 37, SEQ.ID NO: 39, SEQ.ID NO: 41 , SEQ.ID NO: 42, SEQ.ID NO: 44, SEQ.ID NO: 46, SEQ.ID NO: 47, SEQ.ID NO: 49, SEQ.ID NO: 51 , SEQ.ID NO: 52, SEQ.ID NO: 54, SEQ.ID NO: 56, SEQ.ID NO: 57, SEQ.ID NO: 59, SEQ.ID NO: 89, SEQ.ID NO: 91 , SEQ.ID NO: 93, SEQ.ID NO: 95, SEQ.ID NO: 97, SEQ.ID NO: 99, SEQ.ID NO: 207, SEQ.ID NO: 209, SEQ.IDNO: 211 , SEQ.ID NO: 213, SEQ.ID NO: 215, SEQ.ID NO: 217, SEQ.ID NO219, SEQ.ID NO: 221 , SEQ.ID NO: 223, SEQ.ID NO: 225, SEQ.ID NO: 227SEQ.ID NO: 229, SEQ.ID NO: 231 , SEQ.ID NO: 233, SEQ.ID NO: 235,SEQ.ID NO: 237, SEQ.ID NO: 239, SEQ.ID NO: 241 , SEQ.ID NO: 243,SEQ.ID NO: 245, SEQ.ID NO: 247, SEQ.ID NO: 249, SEQ.ID NO: 251 ,SEQ.ID NO: 253, SEQ.ID NO: 255, SEQ.ID NO: 257, SEQ.ID NO: 259,SEQ.ID NO: 261 , SEQ.ID NO: 263, SEQ.ID NO: 265, SEQ.ID NO: 267,1667451850SEQ.ID NO: 269, SEQ.ID NO: 271 , SEQ.ID NO: 273, SEQ.ID NO: 343, or SEQ.ID NO: 345;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.ID NO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO: 216, SEQ.ID NO: 218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224, SEQ.ID NO: 226,SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232, SEQ.ID NO: 234SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240, SEQ.ID NO: 242SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248, SEQ.ID NO: 250SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256, SEQ.ID NO: 258SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264, SEQ.ID NO: 266SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272, SEQ.ID NO: 274SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.ID NO: 35, SEQ.ID NO: 38, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO:1677451850216, SEQ.ID NO: 218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224SEQ.ID NO: 226, SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232,SEQ.ID NO: 234, SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240,SEQ.ID NO: 242, SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248,SEQ.ID NO: 250, SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256,SEQ.ID NO: 258, SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264,SEQ.ID NO: 266, SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272,SEQ.ID NO:

27. , SEQ.ID NO: >44, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

8. A vaccine formulation according to claim 1, wherein the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, is a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 25;(b) a nucleic acid sequence having at least 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with the nucleic acid sequence of (a);(c) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to the nucleic acid sequence of (a);(e) a nucleic acid sequence encoding a polypeptide having the amino acid sequences set forth in SEQ.ID NO: 26, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of the amino acid sequence set forth in SEQ.ID NO: 26, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f); and1687451850the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, is a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 15, or SEQ.ID NO: 17;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 16 or SEQ.ID NO: 18, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 16 or SEQ.ID NO: 18, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

9. A vaccine formulation according to claim 1 , wherein the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, is a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 19, SEQ.ID NO: 21 , SEQ.ID NO: 23, orSEQ.ID NO: 25;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);1697451850(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, orSEQ.ID NO: 26, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, orSEQ.ID NO: 26, or an immunogenically equivalent portion thereto; and(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f); and the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, is a protein expressed by a nucleic acid sequence selected from the group of nucleic acid sequences consisting of:(a) SEQ.ID NO: 9, SEQ.ID NO: 11 , SEQ.ID NO: 13, SEQ.ID NO: 15, SEQ.ID NO: 17, SEQ.ID NO: 89, SEQ.ID NO: 91 , SEQ.ID NO: 93, SEQ.ID NO: 95, SEQ.ID NO: 97, or SEQ.ID NO: 99;(b) a nucleic acid sequence having at least 70% identity with any one of the nucleic acid sequences of (a);(c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) but for the degeneration of the genetic code;(d) a nucleic acid sequence that is complementary to any one of the nucleic acid sequences of (a);(e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, or SEQ.ID NO: 100, or an immunogenically equivalent portion thereof;(f) a nucleic acid sequence that encodes a functional variant of any one of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, or SEQ.ID NO: 100, or an immunogenically equivalent portion thereto; and1707451850(g) a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

10. A vaccine formulation according to claim 1 , wherein the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof comprises any of the amino acid sequences set forth in: SEQ.ID NO: 2, SEQ.ID NO: 4, SEQ.ID NO: 6, SEQ.ID. NO: 8, SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, SEQ.ID NO: 26, SEQ.ID NO: 28, SEQ.ID NO: 30, SEQ.ID NO: 62, SEQ.ID NO: 64, SEQ.ID NO: 66, SEQ.ID NO: 68, SEQ.ID NO: 70, SEQ.ID NO: 72, SEQ.ID NO: 74, SEQ.ID NO: 76, SEQ.ID NO: 78, SEQ.ID NO: 80, SEQ.ID NO: 82, SEQ.ID NO: 84, SEQ.ID NO: 86, SEQ.ID NO: 88, SEQ.ID NO: 102, SEQ.ID NO: 104, SEQ.ID NO: 106, SEQ.ID NO: 108, SEQ.ID NO: 110, SEQ.ID NO: 112 SEQ.ID NO: 114, SEQ.ID NO: 116, SEQ.ID NO: 187, SEQ.ID NO: 120, SEQ.ID NO: 122, SEQ.ID NO: 124, SEQ.ID NO: 126, or SEQ.ID NO: 128, or an immunologically equivalent portion thereof.

11. A vaccine formulation according to claim 1 , wherein the at least one Ngo2TbpB protein, or immunologically equivalent portion thereof, comprises any of the amino acid sequences set forth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.ID NO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.IDNO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55, SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.IDNO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.IDNO: 100, SEQ.ID NO: 208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214,SEQ.ID NO: 216, SEQ.ID NO: 218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224, SEQ.ID NO: 226, SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232, SEQ.ID NO: 234, SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240, SEQ.ID NO: 242, SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248, SEQ.ID NO: 250, SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256, SEQ.ID NO: 258, SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264, SEQ.ID NO: 266, SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272, SEQ.ID NO: 274, SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereof.171745185012. A vaccine formulation according to claim 1 , wherein the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, comprises: any of the amino acid sequences set forth in: SEQ.ID NO: 2, SEQ.ID NO: 4, SEQ.ID NO: 6, SEQ.ID. NO: 8, SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, SEQ.ID NO: 26, SEQ.ID NO: 28, SEQ.ID NO: 30, SEQ.ID NO: 62, SEQ.ID NO: 64, SEQ.ID NO: 66, SEQ.ID NO: 68, SEQ.ID NO: 70, SEQ.ID NO: 72, SEQ.ID NO: 74, SEQ.ID NO: 76, SEQ.ID NO: 78, SEQ.ID NO: 80, SEQ.ID NO: 82, SEQ.ID NO: 84, SEQ.ID NO: 86, SEQ.ID NO: 88, SEQ.ID NO: 102, SEQ.ID NO: 104, SEQ.ID NO: 106, SEQ.ID NO: 108,SEQ.ID NO: 110, SEQ.ID NO: 112 SEQ.ID NO: 114, SEQ.ID NO: 116,SEQ.ID NO: 187, SEQ.ID NO: 120, SEQ.ID NO: 122, SEQ.ID NO: 124,SEQ.ID NO: 126, or SEQ.ID NO: 128, or an immunologically equivalent portion thereof; and the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, comprises: any of the amino acid sequences setforth in SEQ.ID NO: 10, SEQ.IDNO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33,SEQ.ID NO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.IDNO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55,SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.IDNO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO:208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO:216, SEQ.ID NO: 218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO:224, SEQ.ID NO: 226, SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO:232, SEQ.ID NO: 234, SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO:240, SEQ.ID NO: 242, SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO:248, SEQ.ID NO: 250, SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO:256, SEQ.ID NO: 258, SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO:264, SEQ.ID NO: 266, SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO:272, SEQ.ID NO: 274, SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereof.172745185013. A vaccine formulation according to claim 1 , wherein the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, comprises:SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, or SEQ.ID NO: 26, or an immunologically equivalent portion thereof; and the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, comprises: any of the amino acid sequences setforth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.ID NO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.IDNO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55,SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.IDNO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO:208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO:216, SEQ.ID NO: 218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO:224, SEQ.ID NO: 226, SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO:232, SEQ.ID NO: 234, SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO:240, SEQ.ID NO: 242, SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO:248, SEQ.ID NO: 250, SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO:256, SEQ.ID NO: 258, SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO:264, SEQ.ID NO: 266, SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO:272, SEQ.ID NO: 274, SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereof.

14. A vaccine formulation according to claim 1 , wherein the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, comprises SEQ.ID NO: 26, or an immunologically equivalent portion thereof; and the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, comprises: any of the amino acid sequences setforth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 33, SEQ.ID NO: 35, SEQ.ID NO: 38, SEQ.ID NO: 40, SEQ.ID NO: 43, SEQ.ID NO: 45, SEQ.ID NO: 48, SEQ.ID NO: 50, SEQ.ID NO: 53, SEQ.ID NO: 55,1737451850SEQ.ID NO: 58, SEQ.ID NO: 60, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.IDNO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO:208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO:216, SEQ.ID NO: 218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO:224, SEQ.ID NO: 226, SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO:232, SEQ.ID NO: 234, SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO:240, SEQ.ID NO: 242, SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO:248, SEQ.ID NO: 250, SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO:256, SEQ.ID NO: 258, SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO:264, SEQ.ID NO: 266, SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO:272, SEQ.ID NO: 274, SEQ.ID NO: 344, or SEQ.ID NO: 346, or an immunogenically equivalent portion thereof.

15. A vaccine formulation according to claim 1 , wherein the at least one Ngo1 TbpB protein, or immunologically equivalent portion thereof, comprises:SEQ.ID NO: 20, SEQ.ID NO: 22, SEQ.ID NO: 24, or SEQ.ID NO: 26, or an immunologically equivalent portion thereof; and the at least one Ngo2 TbpB protein, or immunologically equivalent portion thereof, comprises: any of the amino acid sequences setforth in SEQ.ID NO: 10, SEQ.ID NO: 12, SEQ.ID NO: 14, SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, or SEQ.ID NO: 100, or an immunogenically equivalent portion thereof.

16. A vaccine formulation according to claim 1 , wherein at least one of the at least one Ngo1 TbpB protein and / or at least one Ngo2 TbpB protein is a modulated TbpB protein, such that the Kd between the modulated TbpB protein and native host iron binding protein exceeds the Kd between the native non-modified TbpB protein and the native host iron binding protein by a factor of at least 2x, at least 10x, or at least 100x, wherein, optionally, the native non-modified TbpB protein and the modulated TbpB protein each comprise at least 100 consecutive amino acid residues which are identical to one another, but for one, two, or three1747451850modulated amino acid residues comprised within the at least 100 consecutive amino acids of the modulated TbpB protein.

17. A vaccine formulation according to claim 16, wherein the modulated TbpB protein is an Ngo1 protein comprising an amino acid sequence selected from SEQ.ID NO: 26, SEQ.ID NO: 62, SEQ.ID NO: 64, SEQ.ID NO: 68, SEQ.ID NO: 70, SEQ.ID NO: 72, SEQ.ID NO: 74, SEQ.ID NO: 76, SEQ.ID NO:80, SEQ.ID NO: 82, SEQ.ID NO: 84, SEQ.ID NO: 86, SEQ.ID NO: 88, SEQ.ID NO; 102, SEQ.ID NO: 108, SEQ.ID NO: 110, SEQ.ID NO: 116, SEQ.ID NO: 122, or SEQ.ID NO: 128, and wherein the Ngo2 protein is a modulated TbpB protein comprising an amino acid selected from SEQ.ID NO: 16, SEQ.ID NO: 18, SEQ.ID NO: 48, SEQ.ID NO: 90, SEQ.ID NO: 92, SEQ.ID NO: 94, SEQ.ID NO: 96, SEQ.ID NO: 98, SEQ.ID NO: 100, SEQ.ID NO: 208, SEQ.ID NO: 210, SEQ.ID NO: 212, SEQ.ID NO: 214, SEQ.ID NO: 216, SEQ.ID NO: 218, SEQ.ID NO: 220, SEQ.ID NO: 222, SEQ.ID NO: 224, SEQ.ID NO: 226, SEQ.ID NO: 228, SEQ.ID NO: 230, SEQ.ID NO: 232, SEQ.ID NO: 234, SEQ.ID NO: 236, SEQ.ID NO: 238, SEQ.ID NO: 240, SEQ.ID NO: 242, SEQ.ID NO: 244, SEQ.ID NO: 246, SEQ.ID NO: 248, SEQ.ID NO: 250, SEQ.ID NO: 252, SEQ.ID NO: 254, SEQ.ID NO: 256, SEQ.ID NO: 258, SEQ.ID NO: 260, SEQ.ID NO: 262, SEQ.ID NO: 264, SEQ.ID NO: 266, SEQ.ID NO: 268, SEQ.ID NO: 270, SEQ.ID NO: 272, SEQ.ID NO: 274, SEQ.ID NO: 344, or SEQ.ID NO: 346.

18. A vaccine formulation according to claim 16, wherein the modulated TbpB protein is an Ngo1 protein comprising an amino acid sequence selected from SEQ.ID NO: 26, SEQ.ID NO: 102, and SEQ.ID NO: 108, and the Ngo2 protein is a modulated TbpB protein comprising an amino acid selected from SEQ.ID NO: 230, SEQ.ID NO: 234, SEQ.ID NO: 238, and SEQ.ID NO: 346.

19. A vaccine formulation according to claim 16, wherein the native nonmodified TbpB protein is an Ngo1 TbpB protein comprising at least 100 consecutive amino acid residues of SEQ.ID NO: 22.

20. A vaccine formulation according to claim 16, wherein the native nonmodified TbpB protein is an Ngo1 TbpB protein comprising at least 1001757451850consecutive amino acid residues of SEQ.ID NO: 22, and the modulated TbpB protein comprises one modulated amino acid residue comprised within the at least 100 consecutive amino acids, the modulated amino acid residue corresponding with aspartic acid amino acid residue 209 (209D) in SEQ.ID NO: 22, and wherein, optionally, aspartic acid amino acid residue 209 (209D) is modulated to replace aspartic acid amino acid residue 209 (209D) in the modulated TbpB protein with a positively charged amino acid residue, optionally, a histidine (H) amino acid residue.

21. A vaccine formulation according to claim 16, wherein the native nonmodified TbpB protein is an Ngo1 TbpB protein comprising at least 100 consecutive amino acid residues of SEQ.ID NO: 22, and the modulated TbpB protein comprises SEQ.ID NO: 26.

22. A vaccine formulation according to claim 1 , wherein the vaccine formulation comprises a fusion polypeptide comprising the first and second N. gonorrhoeae TbpB protein, or immunologically equivalent portion thereof.

23. A vaccine formulation according to claim 1 , wherein the vaccine formulation comprises two N. gonorrhoeae TbpB proteins, or immunogenically equivalent portions thereof.

24. A vaccine formulation according to claim 1 , wherein the vaccine formulation comprises three, four, or five, N. gonorrhoeae TbpB proteins, or immunogenically equivalent portions thereof.

25. A vaccine formulation according to any one of claims 1 to 24, wherein the vaccine formulation is a cross-protective vaccine formulation, wherein the at least two TbpB proteins, or immunogenically equivalent portions thereof, are obtained from a first and second N. gonorrhoeae strain, and the vaccine formulation is for the administration to a human to prevent or ameliorate an infection caused by a strain other than the first or second N. gonorrhoeae strain.176745185026. A vaccine formulation according to any one of claims 1 to 24, wherein the at least two TbpB proteins, or immunogenically equivalent portions thereof, are recombinantly produced in a host cell, optionally a microbial host cell.

27. A vaccine formulation according to any one of claims 1 to 24, wherein the at least two TbpB proteins, or immunologically equivalent portions thereof, are recombinantly produced in a host cell, optionally a microbial host cell, wherein the vaccine formulation is substantially free of host cell constituents, other than the at least two TbpB polypeptides, or immunologically equivalent portions thereof.

28. A vaccine formulation according to claim 25, wherein the infection is caused by an N. gonorrhoeae strain selected from MS11 , FA1090, FAW, WHO F, WHO G, WHO L, WHO M, WHO O, WHO, P, WHO V, WHO W, WHO X, WHO Y.

29. A vaccine formulation according to any one of claims 1 to 28, wherein the vaccine formulation is substantially free of N. gonorrhoeae constituents, other than the at least two TbpB polypeptides, or immunologically equivalent portions thereof.

30. A vaccine formulation according to any one of claims 1 to 29, wherein the vaccine formulation further comprises a pharmaceutically acceptable adjuvant.

31. A vaccine formulation according to claim 30, wherein the vaccine formulation further comprises a pharmaceutically acceptable excipient, carrier, or diluent.

32. A vaccine formulation according to claims 29 or 30, wherein the vaccine formulation comprises from about 0.001 % to about 20% by weight of the at least two TbpB proteins, or the immunogenically equivalent portions thereof, and a pharmaceutically acceptable adjuvant constituting from about 0.1 % to about 60% by weight or volume of the vaccine formulation.

33. A use of a vaccine formulation according to any one of claims 1 to 32, for the prevention, treatment or amelioration of N. gonorrhoeae infection in a human in need thereof.177745185034. A method for preparing a vaccine formulation according to any one of claims 1 to 32, the method comprising:(a) providing a first chimeric nucleic acid sequence comprising as operably linked components:(i) a nucleic acid sequence encoding a first N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, or an mRNA encoding a first N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, wherein the first TbpB protein is an Ngo1 protein; and(ii) one or more nucleic acid sequences capable of controlling expression of the nucleic acid sequence encoding the first TbpB protein, or immunogenically equivalent portion thereof, or the mRNA encoding the first N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, in a host cell;(b) providing a second chimeric nucleic acid sequence comprising as operably linked components:(i) a nucleic acid sequence encoding a second N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, or an mRNA encoding a second N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, wherein the second TbpB protein is an Ngo2 protein; and(ii) one or more nucleic acid sequences capable of controlling expression of the nucleic acid sequence encoding the second TbpB protein, or immunogenically equivalent portion thereof, or an mRNA encoding a first N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, in a host cell;(c) introducing the first and second chimeric nucleic acid sequence each into a separate first and second host cell, or together in a common host cell;(d) growing the first and second separate host cells, or common host cell, to produce the first and second TbpB proteins, or immunogenically equivalent portions thereof, or the mRNA encoding the first or second N.1787451850gonorrhoeae TbpB proteins, or an immunogenically equivalent portion thereof;(e) recovering the first and second TbpB proteins, or immunogenically equivalent portions thereof, or the mRNA encoding the first or second N. gonorrhoeae TbpB proteins, or an immunogenically equivalent portion thereof; and(f) formulating the first and second TbpB proteins, or immunogenically equivalent portions thereof, or the mRNA encoding the first and second N. gonorrhoeae TbpB proteins, or an immunogenically equivalent portion thereof, together with a pharmaceutically acceptable adjuvant to form a vaccine formulation comprising an effective amount of the first and second TbpB protein, or immunogenically equivalent portions thereof, to treat a human susceptible to N. gonorrhoeae infection.

35. A method for preparing a vaccine formulation according to any one of claims 1 to 32, the method comprising:(a) providing a chimeric nucleic acid sequence comprising as operably linked components:(i) a nucleic acid sequence encoding a first N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, or an mRNA encoding a first N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, wherein the first TbpB protein is an Ngo1 TbpB protein;(ii) a nucleic acid sequence encoding a second N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, or an mRNA encoding a second N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, wherein the second TbpB protein is an Ngo2 TbpB protein; and(iii) one or more nucleic acid sequences capable of controlling expression of the nucleic acid sequence encoding the first and the second TbpB protein, or an immunogenically equivalent portion thereof in a host cell;(b) introducing the chimeric nucleic acid sequence into a host cell;1797451850(c) growing the host cell to produce the first and second TbpB proteins, or the immunogenically equivalent portions thereof, or the mRNA encoding the first and second N. gonorrhoeae TbpB proteins, or an immunogenically equivalent portion thereof;(d) recovering the first and second TbpB proteins, or the immunogenically equivalent portions thereof, or the mRNA encoding the first or second N. gonorrhoeae TbpB proteins, or an immunogenically equivalent portion thereof; and(e) formulating the first and second TbpB proteins, or the immunogenically equivalent portions thereof, or the mRNA encoding the first or second N. gonorrhoeae TbpB proteins, or an immunogenically equivalent portion thereof, together with a pharmaceutically acceptable adjuvant to form a vaccine formulation comprising an effective amount of the first and second TbpB protein, or the immunogenically equivalent portions thereof, or the mRNA encoding the first or second N. gonorrhoeae TbpB proteins, or an immunogenically equivalent portion thereof, to treat a human susceptible to N. gonorrhoeae infection.

36. An expression vector comprising a chimeric nucleic acid sequence comprising as operably linked components:(a) a nucleic acid sequence encoding a N. gonorrhoeae TbpB protein, protein, or an immunogenically equivalent portion thereof, or an mRNA encoding a N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof; and(b) a nucleic acid sequence capable of controlling expression of the nucleic acid sequence encoding the N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, or an mRNA encoding a N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, in a host cell, wherein the TbpB protein is an Ngo1 TbpB protein, or an Ngo2 TbpB protein.

37. An expression vector comprising a chimeric nucleic acid sequence comprising as operably linked components:1807451850(a) a nucleic acid sequence encoding a first and second / V. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, or an mRNA encoding a first and second N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof,; and (b) one or more nucleic acid sequences capable of separately or jointly controlling expression of the nucleic acid sequence encoding the first and second N. gonorrhoeae TbpB protein, or the immunogenically equivalent portion thereof, or , or the mRNA encoding the first and second N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, in a host cell, wherein the first and second TbpB protein are an Ngo1 TbpB protein, and an Ngo2 TbpB protein, respectively.

38. A host cell comprising a chimeric nucleic acid comprising as operably linked components:(a) a nucleic acid sequence encoding a N. gonorrhoeae TbpB protein, protein, or an immunogenically equivalent portion thereof, or an mRNA encoding a N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof,; and (b) a nucleic acid sequence capable of controlling expression of the nucleic acid sequence encoding the N. gonorrhoeae TbpB protein, or immunogenically equivalent portion thereof, or the mRNA encoding the N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, in a host cell, wherein the TbpB protein is an Ngo1 TbpB protein or an Ngo2 TbpB protein.

39. A host cell comprising a chimeric nucleic acid sequence comprising as operably linked components:(a) a nucleic acid sequence encoding a first and second N. gonorrhoeae TbpB protein, or immunogenically equivalent portions thereof, or an mRNA encoding a first and second N. gonorrhoeae TbpB protein, or immunogenically equivalent portions thereof,; and(b) one or more nucleic acid sequences capable of separately or jointly controlling expression of the nucleic acid sequence encoding the first1817451850and second N. gonorrhoeae TbpB protein, or the immunogenically equivalent portions thereof, or the mRNA encoding the first and second N. gonorrhoeae TbpB protein, or immunogenically equivalent portions thereof, in a host cell, wherein the first and second TbpB protein are selected from an Ngo1 TbpB protein, and an Ngo2 TbpB protein, respectively.

40. A host cell mixture comprising a first and second host cell, the first host cell comprising a first chimeric nucleic acid comprising as operably linked components:(a) a nucleic acid sequence encoding a first N. gonorrhoeae TbpB protein, protein, or an immunogenically equivalent portion thereof, or an mRNA encoding a first N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof; and (b) a nucleic acid sequence capable of controlling expression of the nucleic acid sequence encoding the first N. gonorrhoeae TbpB protein, or immunogenically equivalent portion thereof, or the mRNA encoding the first N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, in the first host cell, wherein the first TbpB protein is an Ngo1 TbpB protein; and the second host cell comprising a second chimeric nucleic acid comprising as operably linked components:(c) a nucleic acid sequence encoding a second / V. gonorrhoeae TbpB protein, protein, or an immunogenically equivalent portion thereof, or an mRNA encoding a second N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof,; and(d) a nucleic acid sequence capable of controlling expression of the nucleic acid sequence encoding the second N. gonorrhoeae TbpB protein, or immunogenically equivalent portion thereof, or the mRNA encoding the second N. gonorrhoeae TbpB protein, or an immunogenically equivalent portion thereof, in the second host cell, wherein1827451850the second TbpB protein is an Ngo2 TbpB protein.1837451850

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