Modified factor H binding protein

Abstract

The present invention relates to a modified factor H binding protein (fHbp), comprising fHbp or a variant thereof modified by adding at least one exogenous peptide loop; and related nucleic acids, compositions and uses. The present invention also relates to the use of the modified factor H binding protein (fHbp) to treat or prevent pathogenic infection or colonization of a subject.

Description

Technical Field

[0001] The present invention relates to a modified factor H binding protein (fHbp) and its use in eliciting an immune response against pathogenic infection or colonization, such as against Neisseria meningitidis or Neisseria gonorrhoeae. Background Art

[0002] Neisseria meningitidis (Nm) remains the leading cause of sepsis and bacterial meningitis in children and young adults. The onset of illness can be very rapid, and the mortality rate of septicemic illness is approximately 10%. 1 , and those who survive may suffer severe disabilities, including limb loss and neurological defects 1 Therefore, preventive immunization is the best way to protect individuals from meningococcal infection. Vaccines are available in capsule form. 2 , but only partially effective vaccines are available for endemic serogroup B infections, which currently cause more than 80% of cases in the UK 3 The polysaccharide of Neisseria meningitidis (Nm) of serogroup B capsule is poorly immunogenic because it has structural identity with human glycoproteins found in neural tissue and could induce autoimmunity if used as a vaccine. 4 There is therefore an urgent need to generate new vaccines, and tremendous efforts have been made by academia and industry to achieve this important goal.

[0003] PorA is the major target of the immune response to meningococcal outer membrane vesicles (OMVs) 16,17,18 , an integral outer membrane protein (OMP) of meningococci 19 However, the sequence of this protein is diverse, and the prevalence of specific variants varies by geographic region, and OMV vaccines are primarily specific for PorA. Variants of PorA are identified by sequences in the variable region (VR) of the protein, which is located in surface-exposed loops of the protein and is the target of the immune response. 17 PorA has seven extracellular loops; the fourth loop is variable region 2 (VR2) and is the target of most serum bactericidal activity (SBA) produced by PorA following natural infection and immunization with OMVs 16,18 SBAs are known relevant protectors against meningococcal disease. Despite sequence diversity, approximately 70% of UK isolates will be covered by a vaccine containing 6 PorA proteins (http: / / pubmlst.org / neisseria / PorA / ).

[0004] A major obstacle to bacterial vaccine development is the difficulty in producing large amounts of intact OMPs such as PorA in their native conformation. This is because OMPs contain hydrophobic domains that span the membrane and fail to fold properly when expressed as soluble recombinant proteins. Proper folding is critical for PorA because SBAs are triggered by the conformation of the protein rather than by linear epitopes. 20 Previous attempts to use PorA peptides as vaccines have not been successful because they were not sufficiently immunogenic and did not present the immunogenic portion of PorA in its correct conformation. Therefore, the only PorA-based vaccine in development is OMV, which has limited efficacy in infants. 2 , is reactogenic 21 , and are poorly defined, providing regulatory issues. OMVs as immunogens are disadvantageous because consistency and toxicity can be problematic during manufacturing. For example, OMVs can contain toxic lipopolysaccharides (LPS).

[0005] Meningococcal factor H binding protein (fHbp) is a surface-exposed lipoprotein composed of two β-barrels. 5 ( Figure 1A The N-terminal β barrel of fHbp has a relatively open structure, while the C-terminal β barrel is stabilized by extensive hydrogen bonds between the seven β strands that form the barrel. 5 .

[0006] Importantly, fHbp is a key antigen in vaccines against serogroup B Neisseria meningitidis (Nm) being developed by pharmaceutical companies (e.g., Pfizer, GlaxoSmithKline (GSK), etc.) and is included in the next generation of OMV vaccines. 6,7,8 The Pfizer vaccine consists of two fHbps, while the GSK vaccine has a single fHbp in a mixture of other antigens including OMVs. The fHbp binds human (but not murine) factor H (fH) 9,10 , an abundant plasma protein that downregulates the complement system 11 , a key aspect of immunity against Neisseria meningitidis (Nm) 12 Immunization of adolescents and adults with fHbp induces SBA 13 .

[0007] fHbp has been classified into different schemes based on its predicted amino acid sequence. In this application, three variant groups (v1, v2 and v37) and peptide numbers are identified (www.mlst.org). Importantly, sera raised against v1 fHbp do not mediate SBA against N. meningitidis (Nm) expressing v2 and v3 proteins, and vice versa. The GSK vaccine contains a single v1 protein (v1.1), while the Pfizer formulation includes both v1 and v3 fHbp 13,14Therefore, no vaccine currently includes v2fHbp, even though a large proportion of disease in the UK is currently caused by strains expressing fHbp from this variant group. 3,15 .

[0008] WO2011024072 is a patent application describing the use of fHbp, which is selected or engineered to have a sequence that can induce broad-spectrum bactericidal anti-meningococcal antibodies after administration to a host animal. The document teaches that additional meningococcal antigens can be provided by engineering fHbp in the form of N-terminal or C-terminal fusion proteins. However, such proposals are unlikely to produce proteins that can present immunogenic portions of many meningococcal antigens (e.g., PorA) in the correct conformation, and they are not sufficiently immunogenic. Summary of the invention

[0009] It is an object of the present invention to provide an alternative and improved immunogenic molecule for vaccination against pathogenic organisms, in particular for preventing or reducing meningococcal or gonococcal infection or colonization.

[0010] According to a first aspect of the present invention, the present invention provides a modified factor H binding protein (fHbp), which comprises fHbp or a variant thereof modified by adding at least one exogenous peptide loop from a different antigen to serve as a molecular scaffold.

[0011] It has been shown herein that immunogenic peptides (e.g., those from PorA) can be introduced into factor H binding protein (fHbp), which can serve as a molecular scaffold. The peptides introduced into fHbp are presented to the immune system and can elicit a protective response (e.g., SBA). Advantageously, the fHbp molecule provides an ideal molecular scaffold for stabilizing an epitope including a peptide loop to display an epitope, particularly for an epitope that is difficult to stabilize and display in its native conformation, such as a loop from a complete OMP (e.g., PorA). This is in contrast to the teachings in, for example, WO2011024072, in which a simple N-terminal or C-terminal fusion of fHbp and another antigen cannot solve the inherent stability and solubility difficulties of some antigens. In particular, due to the insolubility of its transmembrane domain, many OMPs (e.g., PorA) are difficult to express. PorA has a 16-β-chain barrel structure with surface-exposed loops between chains 1 and 2 (loop 1), chains 7 and 8 (loop 4), chains 9 and 10 (loop 5), and chains 11 and 12 (loop 7), which has been shown to be the most effective antigen. fHbp contains two β-barrels, so the peptide loop sequence from OMP can be inserted into the loop end between the β chains of fHbp, presenting the extracellular loop fragment from the complete OMP in its native conformation for immunization. Therefore, the modified fHbp scaffold molecules of the present invention can be used as preventive or therapeutic vaccines against Neisseria meningitidis (Nm) or gonococci in which a single protein presents key epitopes from two different antigens.

[0012] In one embodiment, the fHbp is a meningococcal fHbp. In another embodiment, the fHbp is a gonococcal fHbp. The fHbp may comprise any of variants v1, v2, and v3. In one embodiment, the fHbp may comprise fHbp v1. In another embodiment, the fHbp may comprise fHbp v2. In another embodiment, the fHbp may comprise fHbp v3.

[0013] In one embodiment, fHbp variant v1 may be variant v1.1, v1.13, v1.14, v1.15, v1.4 or v1.55. In one embodiment, fHbp variant v1 may not be v1.1. In one embodiment, fHbp variant v1 may not be v1.55. In one embodiment, fHbp variant v1 may not be v1.1 or v1.55. In one embodiment, fHbp variant v2 may be variant v2.16, v2.19, v2.22 or v2.25. In one embodiment, fHbp variant v3 may be variant v3.45. In one embodiment, fHbp comprises any one of fHbp variants v1.4, v2.25 or v3.45.

[0014] Variants of fHbp may include orthologs of fHbp. For example, variants of fHbp may include Ghfp, a gonococcal homolog of fHbp. Ghfp is non-functional and closely related to V3fHbp (>95% amino acid (aa) identity with factor H, dissociation constant KD>100 μM).

[0015] In one embodiment, the fHbp to be further modified with at least one exogenous peptide loop may comprise or consist of the following sequence:

[0016]

[0017] In another embodiment, the fHbp to be further modified with at least one exogenous peptide loop may comprise or consist of the following sequence:

[0018]

[0019] In another embodiment, the fHbp to be further modified with at least one exogenous peptide loop may comprise or consist of the following sequence:

[0020]

[0021]

[0022] In another embodiment, the fHbp to be further modified with at least one exogenous peptide loop may comprise or consist of the following sequence:

[0023]

[0024] In another embodiment, the fHbp to be further modified with at least one exogenous peptide loop may comprise or consist of the following sequence:

[0025]

[0026] In another embodiment, the fHbp to be further modified with at least one exogenous peptide loop may comprise or consist of the following sequence:

[0027]

[0028]

[0029] In another embodiment, the fHbp to be further modified with at least one exogenous peptide loop may comprise or consist of the following sequence:

[0030]

[0031] In another embodiment, the fHbp to be further modified with at least one exogenous peptide loop may comprise or consist of the following sequence:

[0032]

[0033] In another embodiment, the fHbp to be further modified with at least one exogenous peptide loop may comprise or consist of the following sequence:

[0034]

[0035] In another embodiment, the fHbp to be further modified with at least one exogenous peptide loop may comprise or consist of the following sequence:

[0036]

[0037] In another embodiment, the fHbp to be further modified with at least one exogenous peptide loop may comprise or consist of the following sequence:

[0038]

[0039] In another embodiment, the fHbp to be further modified with at least one exogenous peptide loop may comprise or consist of the following sequence:

[0040]

[0041] In another embodiment, the Ghfp to be further modified with at least one exogenous peptide loop may comprise or consist of the following sequence:

[0042]

[0043]

[0044] In addition to the exogenous peptide loop provided on the modified fHbp, the variant of the fHbp may comprise one or more amino acid residue mutations (including additions, deletions or substitutions) relative to the wild-type fHbp. For example, the fHbp may serve as a scaffold on which the exogenous peptide loop is provided, and the variant relative to the wild-type may comprise amino acid mutations in the scaffold framework in the region outside the exogenous peptide loop attachment point. Reference to the wild-type fHbp may refer to any variant of the fHbp discussed herein (e.g., any one of SEQ ID NOs: 1-13).

[0045] The variant of fHbp may include at least one amino acid change compared to the amino acid in the wild-type protein. The variant of fHbp may include no more than one amino acid change compared to the wild-type protein. The variant of fHbp may include no more than three amino acid changes compared to the wild-type protein. The variant of fHbp may include no more than four amino acid changes compared to the wild-type protein. The variant of fHbp may include no more than five amino acid changes compared to the wild-type protein. The variant of fHbp may include no more than six amino acid changes compared to the wild-type protein. In one embodiment, a variant of fHbp comprising six amino acid mutations compared to the wild-type protein is provided.

[0046] Amino acid substitutions can be conservative substitutions. For example, the mutated residue can contain properties that are substantially similar to the wild-type substituted residue. For example, the mutated residue can contain a charge or hydrophobicity that is substantially similar or equivalent to the wild-type substituted residue. For example, the substituted residue can contain a molecular weight or steric bulk that is substantially similar to the wild-type substituted residue.

[0047] In one embodiment, variant fHbp may have at least 75% identity with wild type. In another embodiment, variant fHbp may have at least 80% identity with wild type. In another embodiment, variant fHbp may have at least 85% identity with wild type. In another embodiment, variant fHbp may have at least 90% identity with wild type. In another embodiment, variant fHbp may have at least 95% identity with wild type. In another embodiment, variant fHbp may have at least 98% identity with wild type. In another embodiment, variant fHbp may have at least 99% identity with wild type. In another embodiment, variant fHbp may have at least 99.5% identity with wild type. The above percentage variation is not intended to include percentage identity variation (i.e., it is the percentage identity of a separate fHbp component relative to wild type) of adding an exogenous peptide loop, and does not include deletion of fHbp sequence at the site of insertion of a loop from other proteins.

[0048] The modified fHbp can be modified so that it cannot bind to factor H or at least has reduced factor H binding activity. The modified fHbp can be non-functional relative to the function of wild-type fHbp. In one embodiment, the modified fHbp has an impaired ability to bind CFH compared to the wild-type protein, with a KD>2 orders of magnitude. Non-functional fHbp can be provided by mutations in the fHbp sequence. In one embodiment, the non-functional fHbp can be provided by one or more exogenous peptide loops that block the binding site of factor H.

[0049] The amino acid residue mutation can prevent or reduce the complement factor H binding of the modified fHbp. In another embodiment, the amino acid residue mutation may not substantially affect the fHbp function. In one embodiment, the amino acid residue mutation in the fHbp or its variant can be selected from the group consisting of amino acids at positions 85, 133, 134, 135, 136, 204, 206, 211, 212, 213, 222, 225, 227, 231 and 252 on v1.1 fHbp or the corresponding positions of other fHbp.

[0050] In one embodiment, the amino acid residue mutation may comprise or consist of alanine substitution instead of wild-type residue substitution. In one embodiment, the amino acid residue change may comprise or consist of any other amino acid substitution instead of wild-type residue substitution.

[0051] Advantageously, providing a non-functional fHbp (ie, non-binding or low binding of Factor H) may eliminate or reduce any adverse effects of Factor H recruitment on vaccine success.

[0052] In one embodiment, amino acid residue mutation can enhance the stability of the fHbp of modification, particularly in the embodiment that fHbp V2 is provided, fHbp V2 can be stabilized by the mutation in the fHbp V2 sequence. The details of V2 stability mutation can be found in WO2014030003, which is incorporated herein by reference. For example, stabilized amino acid substitution can be one or more amino acids at positions 35, 36, 42, 43, 46, 107, 112, 114, 137 and 138 in fHbp V2. Stabilized substitution can be one or more of Ser35, Leu36, Asp42, Glu43, Arg46, Asp107, Val112, Leu114, Ser137 and Gly138.

[0053] In one embodiment, the exogenous peptide loop is immunogenic.The exogenous peptide loop may be derived from an outer membrane / surface exposed protein.

[0054] The exogenous peptide ring can be of prokaryotic origin. The exogenous peptide ring can be derived from proteins on bacteria, such as the outer membrane protein (OMP) of pathogens. The OMP can be a complete OMP or a lipoprotein. The exogenous peptide ring can be derived from meningococcal proteins, such as meningococcal outer membrane proteins. The exogenous peptide ring can be derived from the outer membrane protein of another pathogen (e.g., Neisseria gonorrhoeae).

[0055] The exogenous peptide loop may comprise a fragment of a transmembrane β-barrel protein. The exogenous peptide loop may comprise a fragment of a β-barrel porin. The exogenous peptide loop may comprise a fragment of PorA. In another embodiment, the exogenous peptide loop may comprise a fragment of FetA.

[0056] The length of the exogenous peptide loop (e.g., PorA fragment) can be 16 amino acids. In one embodiment, the length of the exogenous peptide loop (e.g., PorA fragment) can be 8-20 amino acids. In another embodiment, the length of the exogenous peptide loop (e.g., PorA fragment) can be 8-16 amino acids. In another embodiment, the length of the exogenous peptide loop (e.g., PorA fragment) can be 10-16 amino acids. In another embodiment, the length of the exogenous peptide loop (e.g., PorA fragment) can be 12-16 amino acids. In another embodiment, the length of the exogenous peptide loop (e.g., PorA fragment) can be 14-18 amino acids. In another embodiment, the exogenous peptide loop (e.g., PorA fragment) can be any length sufficient to provide an immunogenic epitope. In another embodiment, the exogenous peptide loop (e.g., PorA fragment) can be any length sufficient to provide an immunogenic epitope and maintain a native conformation relative to a wild-type fragment.

[0057] The exogenous peptide loop can be selected from any one of PorA loops 1-7 or a fragment thereof; and / or a combination thereof. The exogenous peptide loop can be selected from any one of the PorA loops of loop 1 (from between β strands 1 and 2), loop 4 (from between β strands 7 and 8), loop 5 (from between β strands 9 and 10) and loop 7 (from between β strands 11 and 12); or a fragment thereof; and / or a combination thereof.

[0058] The exogenous peptide loop may comprise any peptide selected from PorA loop 1 (between β strands 1 and 2); loop 4 (between β barrels 7 and 8); and loop 5 (between β strands 9 and 10); or fragments thereof; and / or combinations thereof.

[0059] The exogenous peptide loop may comprise PorA loop 1 (between β barrels 1 and 2) or a fragment thereof. The exogenous peptide loop may comprise PorA loop 4 (between β strands 7 and 8) or a fragment thereof. The exogenous peptide loop may comprise PorA loop 5 (between β strands 9 and 10) or a fragment thereof.

[0060] It will be appreciated by the skilled artisan that a variant sequence of the PorA loop may provide minor mutations relative to the wild type and still function as an epitope. Thus, an exogenous peptide loop may comprise a PorA loop variant. A variant may comprise one or more amino acid additions, deletions or substitutions relative to the wild type sequence. In another embodiment, a variant may comprise no more than one amino acid addition, deletion or substitution relative to the wild type sequence. In another embodiment, a variant may comprise no more than 2, 3, 4 or 5 amino acid additions, deletions or substitutions relative to the wild type sequence. Substitutions may be conservative substitutions. For example, alternative amino acid residues having substantially similar properties are provided, such as charge, hydrophobicity, steric size or molecular weight. A variant may comprise a sequence having at least 85% sequence identity to a wild type PorA loop sequence. In another embodiment, a variant may comprise a sequence having at least 90%, 95%, 98%, 99% or 99.5% sequence identity to a wild type PorA loop sequence.

[0061] In one embodiment, the modified fHbp may include two or more exogenous peptide loops. In one embodiment, the modified fHbp may include three or more exogenous peptide loops. In one embodiment, the modified fHbp may include 1-7 exogenous peptide loops. In one embodiment, the modified fHbp may include 1-5 exogenous peptide loops. In one embodiment, the modified fHbp may include 1-3 exogenous peptide loops. In one embodiment, the modified fHbp may include 2-7 exogenous peptide loops. In one embodiment, the modified fHbp may include 3-7 exogenous peptide loops. In one embodiment, the modified fHbp may include 2-5 exogenous peptide loops. In one embodiment, the modified fHbp may include 3-5 exogenous peptide loops. The modified fHbp or its variants may be modified with an exogenous peptide loop at at least one position. The modified fHbp or its variants may be modified with an exogenous peptide loop at at least two positions. The modified fHbp or its variants may be modified with an exogenous peptide loop at at least three positions. The modified fHbp or its variant can be modified with exogenous peptide loops at least four positions. The modified fHbp or its variant can be modified with exogenous peptide loops at least five positions. The modified fHbp or its variant can be modified with exogenous peptide loops at least six positions. The modified fHbp or its variant can be modified with exogenous peptide loops at least seven positions.

[0062] A peptide loop may be provided between the two beta sheets of the factor H binding protein (e.g. using fHbp as a scaffold). The peptide loop may be inserted into one or more amino acid positions in fHbp or a variant thereof, selected from positions between amino acids 49-54, 83-88, 114-124, 199-206, 227-233 and 240-246 of v1.1 fHbp (range including all residues in the loop) into which an exogenous peptide loop (e.g. PorA loop) may be inserted (range including all residues in the loop) or corresponding positions of other fHbp.

[0063] At least one of the exogenous peptide loops may not be provided as an N-terminal or C-terminal fusion.

[0064] The fHbp sequence where the exogenous peptide loop may be inserted may be any of those positions underlined with respect to the fHbp V1.1 primary amino acid sequence below. In embodiments using substitution variant fHBPs, the insertion site may be at an equivalent position.

[0065] Position 1 in fHbp (P1), residues 83-88

[0066] P2, residues 199-206

[0067] P3, residues 227-233

[0068] P4, residues 49-54

[0069] P5, residues 114-124

[0070] P7, residues 240-246

[0071]

[0072] In one embodiment, the insertion site of the exogenous peptide loop may be located at position 1 of fHbp (P1), residues 83-88 (sequence EVDGQL (SEQ ID NO: 15)). In another embodiment, the insertion site of the exogenous peptide loop may be located at position P2, residues 199-206 (sequence KPDGKRHA (SEQ ID NO: 16)). In another embodiment, the insertion site of the exogenous peptide loop may be located at position P3, residues 227-233 (sequence FGGKAQE (SEQ ID NO: 17)). In another embodiment, the insertion site of the exogenous peptide loop may be located at position P4, residues 49-54 (sequence AAQGAE (SEQ ID NO: 18)). In another embodiment, the insertion site of the exogenous peptide loop may be located at position P5, residues 114-124 (sequence IQDSEHSGKM (SEQ ID NO: 19)). In another embodiment, the insertion site of the foreign peptide loop may be located at position P7, residues 240-246 (sequence KTVNGI (SEQ ID NO: 20)).

[0073] The insertion site of the foreign peptide loop at any given position 1-7 can be between any residues identified at positions 1-7 of the fHbp. Alternatively, the insertion site of the foreign peptide loop at any given position 1-7 can be before the first residue or after the last residue identified at positions 1-7 of the fHbp. For example, if the foreign peptide loop is inserted at position 4, the insertion can be *AAQGAE (SEQ ID NO: 21), A*AQGAE (SEQ ID NO: 22), AA*QGAE (SEQ ID NO: 23), AAQ*GAE (SEQ ID NO: 24), AAQG*AE (SEQ ID NO: 25), AAQGA*E (SEQ ID NO: 26) or AAQGAE* (SEQ ID NO: 27), where * indicates the insertion site.

[0074] In another embodiment, it will be appreciated by the skilled person that the exogenous peptide loop insertion site may be variable, so that 1-5 residues in the insertion site region may be removed from fHbp (e.g., replaced by a loop) without significantly affecting the fHbp structure. In one embodiment, one or more amino acid residues at the identification position are replaced / substituted by an exogenous peptide loop. In an alternative embodiment, two, three, four, five or more amino acid residues at the identification position are replaced / substituted by an exogenous peptide loop. In another embodiment, the exogenous peptide loop insertion site may be variable, so that 1-3 residues in the insertion site region may be removed from fHbp (e.g., replaced by a loop) without significantly affecting the fHbp structure. In another embodiment, the exogenous peptide loop insertion site may be variable, so that 1 or 2 residues in the insertion site region may be removed from fHbp (e.g., replaced by a loop) without significantly affecting the fHbp structure. In another embodiment, the exogenous peptide loop insertion site may be variable, so that 1 residue in the insertion site region may be removed from fHbp (e.g., replaced by a loop) without significantly affecting the fHbp structure. In one embodiment, the amino acid residues at the insertion site are replaced by the exogenous peptide loop. In another embodiment, the amino acid residues at the insertion site are replaced by the exogenous peptide loop. In another embodiment, the amino acid residues at the insertion site are replaced by the exogenous peptide loop. In another embodiment, the amino acid residues at the insertion site are replaced by the exogenous peptide loop. In another embodiment, the amino acid residues at the insertion site are replaced by the exogenous peptide loop. In another embodiment, the amino acid residues at the insertion site are replaced by the exogenous peptide loop. In another embodiment, the amino acid residues at the insertion site are replaced by the exogenous peptide loop. In another embodiment, the amino acid residues at the insertion site are replaced by the exogenous peptide loop. In another embodiment, the amino acid residues at the insertion site are replaced by the exogenous peptide loop. In another embodiment, the amino acid residues at the insertion site are replaced by the exogenous peptide loop.

[0075] For example, if the foreign peptide loop is inserted at position 4 and replaces one or more residues, the insertion can be A*QGAE (SEQ ID NO: 28), AA*GAE (SEQ ID NO: 29), AA*AE (SEQ ID NO: 30), AA*E (SEQ ID NO: 31), AA*, A*GAE (SEQ ID NO: 32), A*AE (SEQ ID NO: 33), AA*E (SEQ ID NO: 34), A*E, A*, *AQGAE (SEQ ID NO: 35), *QGAE (SEQ ID NO: 36), *GAE (SEQ ID NO: 37), *AE, *E, AAQ*AE (SEQ ID NO: 38), AAQ*E (SEQ ID NO: 39), AAQ* (SEQ ID NO: 40), AAQG*E (SEQ ID NO: 41), AAQG* (SEQ ID NO: 42), AAQGA* (SEQ ID NO: 43), where * indicates the insertion site and one or more residues are removed from the original sequence.

[0076] The skilled artisan will appreciate that equivalent combinations of insertion sites and / or substitutions of the foreign peptide loops can be made with other alternative sequences identified for insertion positions 1-7.

[0077] In one embodiment, the region of the insertion site can be shifted + / -5 residues upstream or downstream of any position 1-7. Alternatively, the region of the insertion site can be shifted + / -4 residues upstream or downstream of any position 1-7. Alternatively, the region of the insertion site can be shifted + / -3 residues upstream or downstream of any position 1-7. Alternatively, the region of the insertion site can be shifted + / -2 residues upstream or downstream of any position 1-7. Alternatively, the region of the insertion site can be shifted + / -1 residue upstream or downstream of any position 1-7.

[0078] Additionally or alternatively, peptide loops may be provided in positions that sterically prevent fHbp:CFH interactions. These peptide loops include exogenous peptide loops inserted into any one or more of the sites between residues 114-124, 199-206 or 240-246 (e.g., positions 2, 5, and 7), underlined in the V1 primary sequence below:

[0079] 1 CSSGGGGVAA DIGAGLADAL TAPLDHKDKG LQSLTLDQSV RKNEKLKLAA

[0080] 51 QGAEKTYGNG DSLNTGKLKN DKVSRFDFIR QIEVDGQLIT LESGEFQVYK

[0081] 101 QSHSALTAFQ TEQ IQDSEHS GMV AKRQFR IGDIAGEHTS FDKLPEGGRA

[0082] 151 TYRGTAFGSD DAGGKLTYTI DFAAKQGNGK IEHLKSPELN VDLAAADI KP

[0083] 201 DGKR VISGSVLYNQAEKGSYSLGIFGGK AQEVAGSAE V KTVNGI RHIG

[0084] 251 LAAKQ (SEQ ID NO: 44)

[0085] In one embodiment of the invention, the modified fHbp is immunogenic. The modified fHbp can be a recombinant protein. The modified fHbp can be a fusion protein, such as a recombinant fusion protein. The modified fHbp can be an isolated modified fHbp molecule. The modified fHbp molecule of the present invention can be described as a single protein, multivalent vaccine. The modified fHbp can be included in an OMV vaccine.

[0086] In embodiments where more than one exogenous peptide loop is inserted into fHbp or a variant thereof, the exogenous peptide loops may be identical (e.g., identical sequences) or substantially similar. For example, some epitopes (e.g., PorA epitopes) may not elicit sufficient functional responses when displayed alone on fHbp. In this case, the present invention can be used to provide the same epitope at multiple sites on the same modified fHbp molecule, thereby enhancing immunogenic recognition of the epitope.

[0087] Alternatively, the exogenous peptide loops can be different relative to each other. For example, where the exogenous peptide loops are derived from a single protein (e.g., PorA), the different exogenous peptide loops can be derived from different regions of the protein (e.g., PorA). In one embodiment, the different exogenous peptide loops can be derived from overlapping and different regions of the protein (e.g., PorA).

[0088] In embodiments where more than one foreign peptide loop is inserted into fHbp or a variant thereof, the foreign peptide loops may be derived from different species or strains. For example, when a multivalent vaccine is required for multiple different antigens (including different organisms).

[0089] The PorA peptide loop sequence can be selected from any of the sequences provided in Table 1 (e.g., any of SEQ ID NOs: 45-79). A combination of different PorA sequences of Table 1 can be provided in any insertion site P1 to P7 described herein (one loop per site). Additionally or alternatively, two or more identical PorA sequences of Table 1 can be provided in any insertion site P1 to P7 described herein (one loop per site).

[0090] Table 1: Sequences of the PorA VR2 loop, any of which can be inserted into any variant of fHbp to make a chimeric fHbp-PorA protein.

[0091]

[0092]

[0093]

[0094] The skilled artisan will appreciate that variants involving one or two or more amino acid substitutions, additions or deletions may be provided for the PorA sequences of Table 1 without substantially removing the immunogenic function. The substitutions may be similar amino acid residues, for example having similar MW, charge, hydrophobicity or moiety, or synthetic analogs. The skilled artisan will further appreciate that the variants may be truncations of the PorA sequences of Table 1, wherein the truncated variants provide sufficient amino acid residues to form a recognizable epitope. In one embodiment, the PorA sequence has at least 80% identity to any of the sequences in Table 1. In another embodiment, the PorA sequence has at least 85% identity to any of the sequences in Table 1. In another embodiment, the PorA sequence has at least 90% identity to any of the sequences in Table 1. In another embodiment, the PorA sequence has at least 95% identity to any of the sequences in Table 1. In another embodiment, the PorA sequence has at least 98% identity to any of the sequences in Table 1.

[0095] In one embodiment, the fHbp to be further modified with an exogenous peptide loop may comprise or consist of the following sequence:

[0096] Among them, any of the PorA VR2 loop sequences in Table 1 may be substituted for any amino acid or group of amino acids highlighted by a box.

[0097] In one embodiment, the fHbp to be further modified with an exogenous peptide loop may comprise or consist of the following sequence:

[0098] Among them, any of the PorA VR2 loop sequences in Table 1 may be substituted for any amino acid or group of amino acids highlighted by a box.

[0099] In one embodiment, the fHbp to be further modified with an exogenous peptide loop may comprise or consist of the following sequence:

[0100] Among them, any of the PorA VR2 loop sequences in Table 1 may be substituted for any amino acid or group of amino acids highlighted by a box.

[0101] In one embodiment, the fHbp to be further modified with an exogenous peptide loop may comprise or consist of the following sequence:

[0102] Among them, any of the PorA VR2 loop sequences in Table 1 may be substituted for any amino acid or group of amino acids highlighted by a box.

[0103] In one embodiment, the fHbp to be further modified with an exogenous peptide loop may comprise or consist of the following sequence:

[0104] Among them, any of the PorA VR2 loop sequences in Table 1 may be substituted for any amino acid or group of amino acids highlighted by a box.

[0105] In one embodiment, the fHbp to be further modified with an exogenous peptide loop may comprise or consist of the following sequence:

[0106] Among them, any of the PorA VR2 loop sequences in Table 1 may be substituted for any amino acid or group of amino acids highlighted by a box.

[0107] In one embodiment, the fHbp to be further modified with an exogenous peptide loop may comprise or consist of the following sequence:

[0108] Among them, any of the PorA VR2 loop sequences in Table 1 may be substituted for any amino acid or group of amino acids highlighted by a box.

[0109] In one embodiment, the fHbp to be further modified with an exogenous peptide loop may comprise or consist of the following sequence:

[0110] Among them, any of the PorA VR2 loop sequences in Table 1 may be substituted for any amino acid or group of amino acids highlighted by a box.

[0111] In one embodiment, the fHbp to be further modified with an exogenous peptide loop may comprise or consist of the following sequence:

[0112] Among them, any of the PorA VR2 loop sequences in Table 1 may be substituted for any amino acid or group of amino acids highlighted by a box.

[0113] In one embodiment, the fHbp to be further modified with an exogenous peptide loop may comprise or consist of the following sequence:

[0114] Among them, any of the PorA VR2 loop sequences in Table 1 may be substituted for any amino acid or group of amino acids highlighted by a box.

[0115] In one embodiment, the fHbp to be further modified with an exogenous peptide loop may comprise or consist of the following sequence:

[0116] Among them, any of the PorA VR2 loop sequences in Table 1 may be substituted for any amino acid or group of amino acids highlighted by a box.

[0117] In one embodiment, the fHbp to be further modified with an exogenous peptide loop may comprise or consist of the following sequence:

[0118] Among them, any of the PorA VR2 loop sequences in Table 1 may be substituted for any amino acid or group of amino acids highlighted by a box.

[0119] The modified fHbp may comprise the sequence of any one of SEQ ID NOs: 92-109.

[0120] The modified fHbp may comprise a sequence of SEQ ID NOs: 92-109, wherein the sequence YYTKDTNNNLTLVP (SEQ ID NO: 68) is replaced by any PorAVR2 loop sequence (eg, any VR2 loop sequence provided in Table 1).

[0121] In one embodiment, the modified fHbp may comprise or consist of the following sequence:

[0122] or the same sequence with the sequence YYTKDTNNNLTLVP (SEQ ID NO: 68) replaced by any PorA VR2 loop sequence (eg, any of the PorA loop sequences provided in Table 1).

[0123] In one embodiment, the modified fHbp may comprise or consist of the following sequence:

[0124] or the same sequence with the sequence YYTKDTNNNLTLVP (SEQ ID NO: 68) replaced by any PorA VR2 loop sequence (eg, any of the PorA loop sequences provided in Table 1).

[0125] In one embodiment, the modified fHbp may comprise or consist of the following sequence:

[0126] or the same sequence with the sequence YYTKDTNNNLTLVP (SEQ ID NO: 68) replaced by any PorAVR2 loop sequence (eg, any of the PorA loop sequences provided in Table 1).

[0127] In one embodiment, the modified fHbp may comprise or consist of the following sequence:

[0128] or the same sequence with the sequence YYTKDTNNNLTLVP (SEQ ID NO: 68) replaced by any PorAVR2 loop sequence (eg, any of the PorA loop sequences provided in Table 1).

[0129] In one embodiment, the modified fHbp may comprise or consist of the following sequence:

[0130] or the same sequence with the sequence YYTKDTNNNLTLVP (SEQ ID NO: 68) replaced by any PorAVR2 loop sequence (eg, any of the PorA loop sequences provided in Table 1).

[0131] In one embodiment, the modified fHbp may comprise or consist of the following sequence:

[0132] or the same sequence with the sequence YYTKDTNNNLTLVP (SEQ ID NO: 68) replaced by any PorA VR2 loop sequence (eg, any of the PorA loop sequences provided in Table 1).

[0133] In one embodiment, the modified fHbp may comprise or consist of the following sequence:

[0134] or the same sequence with the sequence YYTKDTNNNLTLVP (SEQ ID NO: 68) replaced by any PorA VR2 loop sequence (eg, any of the PorA loop sequences provided in Table 1).

[0135] In one embodiment, the modified fHbp may comprise or consist of the following sequence:

[0136] or the same sequence with the sequence YYTKDTNNNLTLVP (SEQ ID NO: 68) replaced by any PorAVR2 loop sequence (eg, any of the PorA loop sequences provided in Table 1).

[0137] In one embodiment, the modified fHbp may comprise or consist of the following sequence:

[0138] or the same sequence with the sequence YYTKDTNNNLTLVP (SEQ ID NO: 68) replaced by any PorAVR2 loop sequence (eg, any of the PorA loop sequences provided in Table 1).

[0139] In one embodiment, the modified fHbp may comprise or consist of the following sequence:

[0140] or the same sequence with the sequence YYTKDTNNNLTLVP (SEQ ID NO: 68) replaced by any PorAVR2 loop sequence (eg, any of the PorA loop sequences provided in Table 1).

[0141] In one embodiment, the modified fHbp may comprise or consist of the following sequence:

[0142] or the same sequence with the sequence YYTKDTNNNLTLVP (SEQ ID NO: 68) replaced by any PorA VR2 loop sequence (eg, any of the PorA loop sequences provided in Table 1).

[0143] In one embodiment, the modified fHbp may comprise or consist of the following sequence:

[0144]

[0145] or the same sequence with the sequence YYTKDTNNNLTLVP (SEQ ID NO: 68) replaced by any PorAVR2 loop sequence (eg, any of the PorA loop sequences provided in Table 1).

[0146] In one embodiment, the modified fHbp may comprise or consist of the following sequence:

[0147] or the same sequence with the sequence YYTKDTNNNLTLVP (SEQ ID NO: 68) replaced by any PorAVR2 loop sequence (eg, any of the PorA loop sequences provided in Table 1).

[0148] In one embodiment, the modified fHbp may comprise or consist of the following sequence:

[0149] or the same sequence with the sequence YYTKDTNNNLTLVP (SEQ ID NO: 68) replaced by any PorA VR2 loop sequence (eg, any of the PorA loop sequences provided in Table 1).

[0150] In one embodiment, the modified fHbp may comprise or consist of the following sequence:

[0151] or the same sequence with the sequence YYTKDTNNNLTLVP (SEQ ID NO: 68) replaced by any PorAVR2 loop sequence (eg, any of the PorA loop sequences provided in Table 1).

[0152] In one embodiment, the modified fHbp may comprise or consist of the following sequence:

[0153] or the same sequence with the sequence YYTKDTNNNLTLVP (SEQ ID NO: 68) replaced with any PorA VR2 loop sequence (eg, any of the PorA loop sequences provided in Table 1).

[0154] In one embodiment, the modified fHbp may comprise or consist of the following sequence:

[0155] or the same sequence with the sequence YYTKDTNNNLTLVP (SEQ ID NO: 68) replaced with any PorA VR2 loop sequence (eg, any of the PorA loop sequences provided in Table 1).

[0156] In one embodiment, the modified fHbp may comprise or consist of the following sequence:

[0157] or the same sequence with the sequence YYTKDTNNNLTLVP (SEQ ID NO: 68) replaced by any PorAVR2 loop sequence (eg, any of the PorA loop sequences provided in Table 1).

[0158] It will be appreciated by the skilled artisan that one, two, three or four or more amino acid substitutions, deletions or additions may be made to the modified fHbp of the present invention without substantially removing its immunogenic function or affecting its stability. The substitution may be a similar amino acid residue, for example, with a similar MW, charge, hydrophobicity or moiety, or a synthetic analog. These modifications are contemplated as part of the present invention. In one embodiment, the modified fHbp may have at least 75% identity with any modified fHbp described herein. In one embodiment, the modified fHbp may have at least 80% identity with any modified fHbp described herein. In one embodiment, the modified fHbp may have at least 85% identity with any modified fHbp described herein. In one embodiment, the modified fHbp may have at least 90% identity with any modified fHbp described herein. In one embodiment, the modified fHbp may have at least 95% identity with any modified fHbp described herein. In one embodiment, the modified fHbp may be at least 98% identical to any of the modified fHbp described herein. In one embodiment, the modified fHbp may be at least 99% identical to any of the modified fHbp described herein. In one embodiment, the modified fHbp may be at least 99.5% identical to any of the modified fHbp described herein.

[0159] According to another aspect of the present invention, there is provided a nucleic acid which substantially or at least encodes a modified fHbp according to the present invention.

[0160] The nucleic acid can be in a vector (eg, a viral vector).

[0161] According to another aspect of the present invention, there is provided a composition comprising the modified fHbp according to the present invention.

[0162] The composition may include two or more differently modified fHbp molecules according to the present invention (e.g., different forms / species thereof). For example, the composition may include two or more different variants of fHbp modified according to the present invention. For example, the composition may include fHbp variants v1 and v2. The composition may include fHbp variants v2 and v3. In another embodiment, the composition includes at least fHbp variant v2.

[0163] According to another aspect of the present invention, there is provided a composition comprising a nucleic acid according to the present invention.

[0164] The composition may comprise a pharmaceutically acceptable carrier. The composition may further comprise an adjuvant.

[0165] According to another aspect of the present invention, there is provided a modified fHbp, nucleic acid or composition according to the present invention for use as a medicament.

[0166] According to another aspect of the present invention, a modified fHbp, nucleic acid or composition according to the present invention is provided for use in treating or preventing pathogen infection or colonization of a subject.

[0167] According to another aspect of the present invention, there is provided a method for treating or preventing pathogen infection or colonization of a subject, comprising administering to the subject a modified fHbp, nucleic acid or composition according to the present invention.

[0168] According to another aspect of the present invention, there is provided a method of vaccination, comprising administering to the subject a modified fHbp, nucleic acid or composition according to the present invention.

[0169] Administration can be provided in a therapeutically effective amount. A skilled person will be able to determine appropriate dosages and repeated administrations.

[0170] The subject can be a mammal, such as a human.

[0171] The infection may be a bacterial infection. For example, the infection may be meningitis, such as with Neisseria meningitidis or Neisseria gonorrhoeae.

[0172] According to another aspect of the present invention, there is provided a single protein, multivalent vaccine comprising a modified factor H binding protein (fHbp) according to the present invention.

[0173] The vaccine can be used as a preventive or therapeutic vaccine against Neisseria meningitidis (Nm).

[0174] A pharmaceutically acceptable carrier may be used. Additionally or alternatively, an adjuvant may be used. Suitable pharmaceutically acceptable carriers and adjuvants are well known to those skilled in the art.

[0175] According to another aspect of the present invention, there is provided a combination of a modified fHbp according to the present invention and at least one other prophylactically or therapeutically active molecule.

[0176] The at least one other preventive or therapeutically active molecule may comprise a vaccine or antigen different from the modified fHbp according to the invention. The combination may be used in a combination vaccine or therapy. For example, the combination may be used in a combination vaccine or therapy, wherein another meningococcal antigen is provided.

[0177] In one embodiment, the at least one other prophylactically or therapeutically active molecule comprises a monovalent protein:capsule polysaccharide vaccine. The monovalent protein:capsule polysaccharide vaccine may comprise any serogroup C or A capsule with bacterial toxoids, a bivalent vaccine (with serogroup C and A capsular polysaccharides conjugated to bacterial toxoids), a tetravalent (serogroup A, C, Y, W) conjugate vaccine, or a pentavalent (A, C, Y, W, X) conjugate vaccine.

[0178] Alternatively, the at least one other prophylactic or therapeutically active molecule may comprise a conjugate vaccine, wherein an antigen comprising an fHbp scaffold carrying an exogenous peptide loop (e.g., a PorA loop) may be incorporated into the conjugate vaccine as a protein carrier molecule. The conjugate vaccine may comprise any serogroup capsular polysaccharide from A, C, Y, W or X strains, alone or in combination.

[0179] According to another aspect of the present invention, there is provided use of factor H binding protein (fHbp) as an epitope display scaffold.

[0180] Use as an epitope display scaffold may comprise the use of a Factor H binding protein (fHbp) comprising any of the modifications described herein.

[0181] In addition to their potential use as vaccines, the compositions or modified fHbp according to the invention may be used as diagnostic reagents and as a measure of vaccine immunocompetence.

[0182] The term "immunogenic" means that the molecule is capable of eliciting an immune response in a human or animal. The immune response may be protective.

[0183] The immune response triggered by the modified fHbp of the present invention may affect the ability of Neisseria meningitidis (Nm) to infect a subject immunized with the modified fHbp of the present invention. Preferably, the ability of Neisseria meningitidis (Nm) to infect a subject immunized with the modified fHbp of the present invention is hindered or prevented. The triggered immune response can recognize and destroy Neisseria meningitidis (Nm). Alternatively or additionally, the triggered immune response can hinder or prevent the replication of Neisseria meningitidis (Nm). Alternatively or additionally, the triggered immune response can hinder or prevent Neisseria meningitidis (Nm) from causing disease in humans or non-human animals.

[0184] As used herein, the term "peptide loop" refers to a single-chain polypeptide sequence anchored at both ends (eg, anchored to a scaffold (eg, fHbp)). The term "loop" does not infer or require any particular secondary structure adopted by the polypeptide.

[0185] The term "exogenous" used in the context of "exogenous peptide loop" should be understood to mean that the peptide loop originates from a different source relative to the fHbp protein (i.e., it is not fHbp or a fragment thereof). However, it may be from the same organism as fHbp. For example, the modified fHbp may include a Neisseria meningitidis fHbp modified with a (exogenous) peptide loop derived from Neisseria meningitidis PorA.

[0186] The term "fusion protein" as used herein should be understood to refer to a polypeptide comprising a combination of sequences from different gene products or sources. The term "fusion protein" can be used interchangeably with the term "chimeric molecule".

[0187] Reference to sequence "identity" as used herein may refer to the percent identity between two aligned sequences using standard NCBI BLASTp parameters (http: / / blast.ncbi.nlm.nih.gov).

[0188] When applied to the modified fHbp of the present invention, the term "isolated" refers to a protein that is: (i) encoded by a nucleic acid using recombinant DNA methods or viral vectors; or (ii) synthesized, for example, by chemical synthesis methods; or (iii) separated from biological materials and then purified. The isolated polypeptides of the present invention include proteins expressed by a nucleotide sequence encoding the protein, or proteins expressed by a recombinant vector containing a nucleotide sequence encoding the protein.

[0189] The term "protective" means preventing disease, reducing the risk of disease infection, transmission and / or progression, reducing the severity of disease, curing a condition or disease, alleviating symptoms, or reducing the severity of disease or disease symptoms.

[0190] The term "prophylactic" refers to a preventive or protective treatment against a disease. Prophylactic may include reducing the risk of infection, transmission and / or progression, or reducing the severity of a disease.

[0191] The term "treat" or "treating" refers to curing a condition or disease, alleviating the symptoms, or reducing the severity of a disease or disease symptoms.

[0192] The skilled person will appreciate that optional features of one embodiment or aspect of the invention may be applicable to other embodiments or aspects of the invention where appropriate. BRIEF DESCRIPTION OF THE DRAWINGS

[0193] Embodiments of the invention will now be described in more detail, by way of example only, with reference to the accompanying drawings.

[0194] Figure 1: Design of the chimeric fHbp:PorA

[0195] (A) Schematic representation of the surface of N. meningitidis showing the dominant outer membrane features, lipooligosaccharides and type 4 fimbriae, and the important antigens, fHbp and PorA. The immunogenic PorA VR2 loop is highlighted, and the interaction of fHbp with domains 6 and 7 of human CFH is shown.

[0196] (B) Structure of V1 fHbp with CFH domains 6-7 showing the six positions (P1-5 and P7) used to generate the chimeric fHbp:PorA that has been inserted into the PorA loop. Note: Position 5 is in the fHbp:CFH interface.

[0197] Figure 2: Using fHbp as a molecular scaffold

[0198] A) Protein structure of fHbp V1.1 (represented by grey ribbons) with six positions (P1-5 and P7) used to generate chimeric fHbp:PorA. B) Secondary structure of fHbp V1.1 (grey arrows indicate beta sheets, rectangles indicate alpha helices). The location of the VR2 P1.16PorA insertion site is indicated by a solid black line, and the numbers indicate the residue ranges that can be inserted into the PorA VR2 loop (corresponding to the residue numbers in ID). C) Purified fHbp:PorA with the PorA P1.16 VR2 loop at positions 1-5 or 7 of wild-type (WT V1.1) fHbp were analyzed by SDS-PAGE and Western blotting. The blots were probed with α-V1 fHbp pAb and α-P1.16 monoclonal antibody (mAb). D) Primary sequence of V1.1 fHbp (SEQ ID NO: 1) indicating the location of positions 1-5 and 7 where loops from other proteins can be inserted (underlined).

[0199] Figure 3: Characteristics of the chimeric fHbp:PorA

[0200] (A) Structure of fHbp:PorA superimposed with the P1.16 loop (black, PDBID: 2mpa), Fab of α-P1.16 monoclonal antibody (mAb) and fHbp:PorA with the loops at positions 1, 3 and 7, demonstrating that the epitope is in a conformation recognized by bactericidal antibodies.

[0201] (B) Stability of the fHbp N-terminal (NT) and C-terminal (CT) β-barrels of fHbp by differential scanning calorimetry using a 20-120°C temperature gradient. TM ) and C-terminus (CT TM) barrel. Binding of fHbp:PorA to complement factor H (CFH) and monoclonal antibody (mAb) SPR analysis of fHbp and chimeric fHbp:PorA coupled to a BIAcore CM5 chip. CFH (fH67) was flowed through at a dilution range of 0.5-32 nM and the dissociation constant (K D ); fHbp: PorA dissociation constant (K D ) confirmed the lack of CFH binding to the loop at position 5 of fHbp, which affects the fHbp:CFH interface ( Figure 2A ). NB = no binding.

[0202] Figure 4: Selection of antigen library for chimeric fHbp:PorA

[0203] (A, B) Frequencies of fHbp variants and PorAVR2 isoforms in UK meningitidis disease isolates from 2010-2015 from the Meningitis Research Foundation Genome Library (http: / / www.meningitis.org / research / genome), shown as pie charts (top) and tables (bottom) with frequencies of specific fHbps and PorAs, respectively.

[0204] Note: V2 fHbp accounted for 38.9% of the isolates.

[0205] Table 2. Accurate sequence matches for all UK isolates of Neisseria meningitidis between 2009 and 2015:

[0206]

[0207] Exact sequence match:

[0208] -Pfizer vaccine, 3.02%

[0209] -Bexsero fHbp (V1.1) or PorA (P1.16), 15.86%

[0210] - Chimeric fHbp: PorA, fHbp or PorA, 72%

[0211] (23.5% of fHbp and PorA)

[0212] Figure 5: Recognition of N. meningitidis fHbp (A) and PorA (B) antigens by mouse immune sera. Whole cell lysates from N. meningitidis strains H44 / 76 (WT), H44 / 67ΔfHbp and H44 / 67ΔPorA were separated by SDS-PAGE, transferred to PVDF membranes and probed with mouse immune sera. Mouse immune sera were obtained by immunizing BalbC mice three times with 20 μg of purified fHbp-PorA chimeras with VR2 loops of P1, P2, P4, P5 or P7.

[0213] Figure 6: Stabilization of V2 fHbp: construct and immunogenicity of fHbp:PorA

[0214] (A) Stabilization of V2 fHbp V2.22 and V2.25 with six (M6) or two (M2) amino acid substitutions. DSC analysis was performed with 20 μM of protein using a temperature gradient of 20-120°C. The fHbp N-terminal barrel (NT TM ) and C-terminal barrel (CT TM ) melting temperature.

[0215] (B) Chimeric fHbp:PorA, the PorA loop is recognized by the corresponding monoclonal antibody (mAb). The PorA VR2 loop from P1.2, P1.4, P1.9, P1.14 and P1.15 was inserted into position 1 of V2.25 fHbp. SDS-PAGE and Western blot analysis of purified wild-type V2.25 fHbp and V3.45 fHbp-PorA chimeras. Western blots were probed with fHbp V2.25 pAb and loop-specific PorA monoclonal antibody (mAb) (NIBSC). CFH binding was detected by long-range Western blot analysis with normal human serum and CFH pAb.

[0216] Figure 7 : Chimeric fHbp:PorA elicits protective immunity. Mice were immunized three times with chimeric fHbp:PorA (i.e., P(1)(1)(13), P(2)(1)(13), and P(3)(1)(13)) and SBAs were measured against the indicated strains; SBAs > 8 were considered protective. The lack of a PorA-directed SBA (i.e., SBA 0, against fHbp mutants) with the VR2 loop at position 3 (i.e., P(3)(1)(13)) is likely because this loop does not protrude from the fHbp barrel as far as position 1.

[0217] Figure 8: Frequency of PorA VR2 (A) and fHbp variants (B) in N. meningitidis serogroup B strains (n=243) isolated in the UK in 2016. Data downloaded from the Meningococcal Research Foundation on 27 June 2017. Other: remaining alleles occurring in <4 isolates. (C) Analysis of recombinant chimeric antigens by SDS-PAGE and Western blotting. Immunoblots probed with α-PorA VR2 monoclonal antibodies (mAbs): P1.4, P1.9, P1.14 and P1.15. (D) Recombinant chimeric antigens were expressed by 1:1% fHbp from the chimeric antigen fHbp. V1.4 :PorA 151 / P1.1.10_1 、fHbp V1.4 :PorA 151 / P1.14 and fHbp V1.4 :PorA 151 / P1.15 、fHbp V3.45 :PorA 158 / P1.4 and fHbp V3.45 :PorA 158 / P1.9 Mouse polyclonal antiserum detected PorA in a panel of Neisseria meningitidis serogroup B isolates. DETAILED DESCRIPTION

[0218] Example 1

[0219] It has been shown that immunogenic peptides can be introduced into factor H binding protein (fHbp) and presented to the immune system and are able to elicit a protective response (Figures 5 and Figure 7 ). Peptides from the integral membrane protein PorA have been used for proof of principle of this method. PorA is difficult to express due to the insolubility of its transmembrane domain. The immunogenic part of the molecule is present in the extracellular loops that are exposed to the immune system. However, an effective immune response is only generated against the loops in their correct conformation; linear peptide sequences do not elicit a functional immune response. By understanding the structure of fHbp, the PorA loop can be introduced into fHbp and generate a relevant response against PorA. This leads to chimeric molecules, based on fHbp and PorA sequences in specific sites to generate chimeric molecules. This method can be used for any other immunogenic intact outer membrane protein.

[0220] Possible reasons for the exclusion of v2fHbp from vaccines have been suggested to be the inherent instability of its N-terminal β-barrel: i) The atomic structure of this part of v2fHbp could not be determined 10 , ii) v2fHbp is sensitive to protease digestion (mass spectrometry demonstrated that the cleavage site is present in the N-terminal β barrel, not shown), and iii) differential scanning calorimetry confirmed that the instability is located in this region of v2fHbp 10 .

[0221] Stable v2 fHbp has been successfully generated. Mutagenesis affecting the N-terminal barrel has been performed, replacing amino acids (aas) alone or in combination. Substitution of six amino acids in M6 fHbp stabilized v2 fHbp (i.e. six changes in the c 130 amino acid substitutions of the β barrel, <0.5%) (see WO2014030003 for details on the mutations). This was visible from differential scanning calorimetry (DSC) and protease sensitivity (see WO2014030003 for details). The side chains of the altered residues promote interactions between the β folds of the N-terminal barrel and are therefore towards the center of the molecule; these changes did not affect the immunogenicity of the protein as expected (no difference in SBA, or no α-fHbp IgG levels were shown).

[0222] A chimeric v1.1 fHbp has been generated that incorporates the 13 amino acids VR2 from P1.16PorA that elicits SBA in recipients of the OMV vaccine 16 While integral membrane proteins contain a hydrophobic (and therefore insoluble) β-barrel, fHbp contains two barrels that can be expressed and purified to high levels. VR2 sequences have been introduced into six different positions of fHbp ( Figure 2B ); these sites were chosen based on similar spacing of flanking β-sheets in PorA22 and fHbp to reduce the likelihood that the insertion would disrupt the overall structure of fHbp ( Figure 2B is the predicted effect of the insertion). The immunogenicity of three fHbps with VR2 inserted into different fHbps has been evaluated ( Figure 2B All proteins elicited antibody responses against fHbp and PorA (Figures 5 and Figure 7 ), and importantly, both proteins tested so far (VR2 at either site 1 or site 2) independently elicited SBAs against fHbp and PorA (SBA of 512 for N. meningitidis (Nm) H44 / 76 and SBA of 256 for both fHbps for N. meningitidis (Nm) H44 / 76αfHbp), providing proof of principle for this approach.

[0223] fHbp as a scaffold for multivalent vaccines

[0224] Nonfunctional fHbp as a vaccine - When clinical trials of vaccines containing fHbp began, the function of fHbp was unknown; fHbp exhibited a high affinity interaction with fH (dissociation constant <5 nM) that was independent of the variant group, and fH engaged a large region of fHbp 5This interaction may affect the use of fHbp as a vaccine by: i) blocking immunogenic epitopes and preventing the production of antibodies that may compete with fH, and ii) reducing complement activation (through fH recruitment) and thus B cell activation at the site of immune induction. Use of a non-functional fHbp could avoid these problems. Key amino acids of v1, v2, and v3 fHbp have been identified as being required for fH interaction 10 , and have shown that modification of v1, v2, and v3 fHbp with single amino acid substitutions that prevent fH binding can preserve or even enhance the immunogenicity of this important vaccine antigen 10,23 .

[0225] To generate and evaluate single protein, multivalent vaccine candidates:

[0226] Neisseria meningitidis (Nm) 15 Protective PorA epitopes for prevalent serosubtypes of the United States (defined by their PorA sequences) were introduced into v1, the stabilized v2 and v3 proteins; their stability and recognition by PorA and fHbp monoclonal antibodies (mAbs) have been determined. PorA sequences have been chosen to cover the diversity of UK isolates, but data from any collection of meningococcal strains can be used.

[0227] Research Methods

[0228] Generation and characterization of vaccine candidates - construction of recombinant fHbp and expression in E. coli using standard plasmid vectors; use of polyHis tag in protein, anion exchange and gel filtration of chimeric v1, V2 and V3 fHbp 10 The proteins were affinity purified because they are either existing vaccines (v1.1 and v3.45) or because of experience with the proteins (v2) or because of their prevalence in N. meningitidis (Nm) strains. The PorA loop has been introduced into fHbp by standard methods, and we have demonstrated that the fusion protein binds to a monoclonal antibody (mAb) against common serotypes of PorA. This strategy has been used to compare native and designed sequences and to fine-tune the antigenicity and fH binding of the candidates. DSC has been performed using VP capillary DSC (GE Healthcare) and SPR with a Biacore 3000 (GE Healthcare) or ProteOn XPR36 (BioRad) as described previously. 10 .

[0229] References

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[0241] 12. Schneider, M.C., Exley, R.M., Ram, S., Sim, R.B. and Tang, C.M. 2007. Interactions between Neisseria meningitidis and the complement system. Trends Microbiol 15: 233 - 240

[0242] 13. Richmond, P.C., Marshall, H.S., Nissen, M.D., Jiang, Q. et al. 2012. Safety, immunogenicity, and tolerability of meningococcal serogroup B bivalent recombinant lipoprotein 2086 vaccine in healthy adolescents: a randomised, single - blind, placebo - controlled, phase 2 trial. Lancet Infect Dis. 12: 597 - 607.

[0243] 14. Gorringe AR, Pajón R 2011. Bexsero: a multicomponent vaccine for prevention of meningococcal disease. Expert Opin Biol Ther. 11: 969 - 85.

[0244] 15. Lucidarme, J., Comanducci, M., Findlow, J., Gray, S. J., et al. 2010. Characterization of fHbp, nhba (gna2132), nadA, porA, and sequence type in group B meningococcal case isolates collected in England and Wales during January 2008 and potential coverage of an investigational group B meningococcal vaccine. Clin Vaccine Immunol. 2010 17:919 - 29

[0245] 16. Rosenqvist, E., E. A., Wedege, E., Caugant, D. A., et al. 1993. A new variant of serosubtype P1.16 in Neisseria meningitidis from Norway, associated with increased resistance to bactericidal antibodies induced by a serogroup B outer membrane protein vaccine. Microb Pathog. 15:197

[0246] 17. Martin, S. L., Borrow, R., van der Ley, P., Dawson, M., Fox, A. J., and Cartwright, K. A. 2000. Effect of sequence variation in meningococcal PorA outer membrane protein on the effectiveness of a hexavalent PorA outer membrane vesicle vaccine. Vaccine 18:2476 - 81.

[0247] 18. Martin, D. R., Ruijne, N., McCallum, L., O'Hallahan, J., and Oster, P. 2006. The VR2 epitope on the PorA P1.7-2,4 protein is the major target for the immune response elicited by the strain-specific group B meningococcal vaccine MeNZB. Clinical and Vaccine Immunology 13:486-491.

[0248] 19. McGuinness, B., Barlow, A. K., Clarke, I. N., Farley, J. E., et al. 1990 Deduced amino acid sequences of class 1 protein (PorA) from three strains of Neisseria meningitidis. Synthetic peptides define the epitopes responsible for serosubtype specificity. J Exp Med. Jun 171:1871-82.

[0249] 20. Christodoulides, M., McGuinness, B. T., Heckels, J. E. 1993. Immunization with synthetic peptides containing epitopes of the class 1 outer-membrane protein of Neisseria meningitidis: production of bactericidal antibodies on immunization with a cyclic peptide. J Gen Micro 139:1729

[0250] 21. Gossger, N., Snape, M.D., Yu, L.M., Finn, A. et al. 2012. Immunogenicity and tolerability of recombinant serogroup B meningococcal vaccine administered with or without routine infant vaccinations according to different immunization schedules: a randomized controlled trial. JAMA. 307:573 - 82.

[0251] 22. van den Elsen, J.M.H., Herron, J.N., Hoogerhout, P., Poolman, J.T. et al. 1997. Bactericidal antibody recognition of a PorA epitope of Neisseria meningitidis: Crystal structure of a Fab fragment in complex with a fluorescein - conjugated peptide. Proteins: Structure, Function, and Bioinformatics 29:113 - 125.

[0252] 23. Beernink, P.T., Shaughnessy, J., Braga, E.M., Liu, Q. et al. 2011. A meningococcal factor H binding protein mutant that eliminates factor H binding enhances protective antibody responses to vaccination. J. Immunol. 186:3606 - 3614.

[0253] 24. Ufret-Vincenty, RL, Aredo, B., Liu,

[0254] All references are incorporated herein by reference.

[0255] Example 2 - Chimeric antigens containing an expanded range of the PorA VR2 loop generate immune responses

[0256] To test the adaptability of the fHbp:PorA chimeric antigens, several chimeric antigens consisting of different combinations of fHbp and PorA VR2 were generated. The comprehensive meningococcal genomic data available for strains isolated in the UK (the Meningitis Research Foundation Meningococcal Genomic Library, developed in collaboration between Public Health England, the Wellcome Trust Sanger Institute and the University of Oxford) enabled the construction of chimeric antigens with exact sequence matches to the most common antigens in a given region. In 2016, the most prevalent PorA VR2s in serogroup B N. meningitidis isolates were P1.4 (15.2%), P1.14 (15.2%), P1.9 (12.8%), P1.16 (11.1%) and P1.15 (5.8%). Figure 8B ). VR2 P1.10_1 was present in 1.6% of serogroup B isolates. The most prevalent variant 1, variant 2, and variant 3 fHbp were V1.4, V2.19, and V3.45, present in 21.8%, 5.3%, and 4.9% of serogroup B N. meningitidis isolates, respectively ( Figure 8C Five different chimeric antigens were constructed, in which PorA VR2 was inserted at position 151 (V1.4) or position 158 (V3.45, Fig. 8A After chimeric antigen expression and purification, Western blot analysis confirmed that these chimeric antigens retained the epitopes recognized by their cognate α-VR2 monoclonal antibody (mAb) and α-fHbppAb ( Fig.8D The thermal stability of wild-type fHbp V1.1, V1.4 and V3.45 and the chimeric antigen was determined by differential scanning calorimetry (DSC, Table 3).

[0257] To test the ability of these fHbp:PorA chimeric antigens to elicit an immune response, groups of CD1 mice were immunized with each chimeric antigen / alum; antisera obtained following immunization were pooled. To assess the resulting PorA immune response, Western blotting was performed using the pooled antisera and a panel of serogroup B Neisseria meningitidis disease isolates. Fig. 8E All chimeric antigens were demonstrated to elicit α-PorA antibodies that recognized their cognate PorA VR2. To assess α-PorA SBA responses, serum bactericidal assays were performed with pooled chimeric antigen / alum antisera and serogroup B N. meningitidis strains with mismatched fHbp variants to negate fHbp cross-protection. Titers ranged from ≥20 to ≥1280 and were above the ≥8 threshold for acceptable correlation with protective immunity against N. meningitidis (Andrews, N. et al. Clin Diagn Lab Immunol 10, 780-786 (2003)) (Table 4).

[0258] Table 3: Stability of wild-type fHbp and chimeric antigens

[0259]

[0260]

[0261] Melting temperature, T m

[0262] Table 4: Serum bactericidal titers

[0263]

[0264] α-PorA SBA titers were generated using pooled chimeric antigen / alum antisera and serogroup B N. meningitidis isolates with mismatched fHbp variants. *fHbp is truncated at residue 242.

[0265] fHbp not tested V1.4 :PorA 151 / P1.15 , as it is required to generate the ΔfHbp strain, as the fHbp in the strain with PorA VR2P1.15 is not mismatched. Sequence Listing <110> Oxford Innovations Ltd <120> Modified factor H binding protein <130> JA69120P.WOP <150> GB1614687.0 <151> 2016-08-31 <160> 109 <170> PatentIn version 3.5 <210> 1 <211> 255 <212> PRT <213> Neisseria meningitidis <400> 1 Cys Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu 1 5 10 15 Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp Lys Gly Leu Gln 20 25 30 Ser Leu Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu 35 40 45 Ala Ala Gln Gly Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn 50 55 60 Thr Gly Lys Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile Arg 65 70 75 80 Gln Ile Glu Val Asp Gly Gln Leu Ile Thr Leu Glu Ser Gly Glu Phe 85 90 95 Gln Val Tyr Lys Gln Ser His Ser Ala Leu Thr Ala Phe Gln Thr Glu 100 105 110 Gln Ile Gln Asp Ser Glu His Ser Gly Lys Met Val Ala Lys Arg Gln 115 120 125 Phe Arg Ile Gly Asp Ile Ala Gly Glu His Thr Ser Phe Asp Lys Leu 130 135 140 Pro Glu Gly Gly Arg Ala Thr Tyr Arg Gly Thr Ala Phe Gly Ser Asp 145 150 155 160 Asp Ala Gly Gly Lys Leu Thr Tyr Thr Ile Asp Phe Ala Ala Lys Gln 165 170 175 Gly Asn Gly Lys Ile Glu His Leu Lys Ser Pro Glu Leu Asn Val Asp 180 185 190 Leu Ala Ala Ala Asp Ile Lys Pro Asp Gly Lys Arg His Ala Val Ile 195 200 205 Ser Gly Ser Val Leu Tyr Asn Gln Ala Glu Lys Gly Ser Tyr Ser Leu 210 215 220 Gly Ile Phe Gly Gly Lys Ala Gln Glu Val Ala Gly Ser Ala Glu Val 225 230 235 240 Lys Thr Val Asn Gly Ile Arg His Ile Gly Leu Ala Ala Lys Gln 245 250 255 <210> 2 <211> 255 <212> PRT <213> Neisseria meningitidis <400> 2 Cys Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu 1 5 10 15 Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp Lys Ser Leu Gln 20 25 30 Ser Leu Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu 35 40 45 Ala Ala Gln Gly Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn 50 55 60 Thr Gly Lys Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile Arg 65 70 75 80 Gln Ile Glu Val Asp Gly Gln Leu Ile Thr Leu Glu Ser Gly Glu Phe 85 90 95 Gln Val Tyr Lys Gln Ser His Ser Ala Leu Thr Ala Leu Gln Thr Glu 100 105 110 Gln Val Gln Asp Ser Glu His Ser Gly Lys Met Val Ala Lys Arg Gln 115 120 125 Phe Arg Ile Gly Asp Ile Ala Gly Glu His Thr Ser Phe Asp Lys Leu 130 135 140 Pro Glu Gly Gly Arg Ala Thr Tyr Arg Gly Thr Ala Phe Gly Ser Asp 145 150 155 160 Asp Ala Ser Gly Lys Leu Thr Tyr Thr Ile Asp Phe Ala Ala Lys Gln 165 170 175 Gly His Gly Lys Ile Glu His Leu Lys Ser Pro Glu Leu Asn Val Asp 180 185 190 Leu Ala Ala Ser Asp Ile Lys Pro Asp Lys Lys Arg His Ala Val Ile 195 200 205 Ser Gly Ser Val Leu Tyr Asn Gln Ala Glu Lys Gly Ser Tyr Ser Leu 210 215 220 Gly Ile Phe Gly Gly Gln Ala Gln Glu Val Ala Gly Ser Ala Glu Val 225 230 235 240 Glu Thr Ala Asn Gly Ile Arg His Ile Gly Leu Ala Ala Lys Gln 245 250 255 <210> 3 <211> 255 <212> PRT <213> Neisseria meningitidis <400> 3 Cys Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu 1 5 10 15 Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp Lys Gly Leu Gln 20 25 30 Ser Leu Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu 35 40 45 Ala Ala Gln Gly Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn 50 55 60 Thr Gly Lys Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile Arg 65 70 75 80 Gln Ile Glu Val Asp Gly Lys Leu Ile Thr Leu Glu Ser Gly Glu Phe 85 90 95 Gln Val Tyr Lys Gln Ser His Ser Ala Leu Thr Ala Leu Gln Thr Glu 100 105 110 Gln Val Gln Asp Ser Glu Asp Ser Gly Lys Met Val Ala Lys Arg Gln 115 120 125 Phe Arg Ile Gly Asp Ile Ala Gly Glu His Thr Ser Phe Asp Lys Leu 130 135 140 Pro Lys Gly Gly Ser Ala Thr Tyr Arg Gly Thr Ala Phe Gly Ser Asp 145 150 155 160 Asp Ala Gly Gly Lys Leu Thr Tyr Thr Ile Asp Phe Ala Ala Lys Gln 165 170 175 Gly His Gly Lys Ile Glu His Leu Lys Ser Pro Glu Leu Asn Val Glu 180 185 190 Leu Ala Thr Ala Tyr Ile Lys Pro Asp Glu Lys Arg His Ala Val Ile 195 200 205 Ser Gly Ser Val Leu Tyr Asn Gln Asp Glu Lys Gly Ser Tyr Ser Leu 210 215 220 Gly Ile Phe Gly Gly Gln Ala Gln Glu Val Ala Gly Ser Ala Glu Val 225 230 235 240 Glu Thr Ala Asn Gly Ile His His Ile Gly Leu Ala Ala Lys Gln 245 250 255 <210> 4 <211> 255 <212> PRT <213> Neisseria meningitidis <400> 4 Cys Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu 1 5 10 15 Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp Lys Ser Leu Gln 20 25 30 Ser Leu Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu 35 40 45 Ala Ala Gln Gly Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn 50 55 60 Thr Gly Lys Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile Arg 65 70 75 80 Gln Ile Glu Val Asp Gly Gln Leu Ile Thr Leu Glu Ser Gly Glu Phe 85 90 95 Gln Val Tyr Lys Gln Ser His Ser Ala Leu Thr Ala Leu Gln Thr Glu 100 105 110 Gln Glu Gln Asp Pro Glu His Ser Gly Lys Met Val Ala Lys Arg Arg 115 120 125 Phe Lys Ile Gly Asp Ile Ala Gly Glu His Thr Ser Phe Asp Lys Leu 130 135 140 Pro Lys Asp Val Met Ala Thr Tyr Arg Gly Thr Ala Phe Gly Ser Asp 145 150 155 160 Asp Ala Gly Gly Lys Leu Thr Tyr Thr Ile Asp Phe Ala Ala Lys Gln 165 170 175 Gly His Gly Lys Ile Glu His Leu Lys Ser Pro Glu Leu Asn Val Glu 180 185 190 Leu Ala Thr Ala Tyr Ile Lys Pro Asp Glu Lys His His Ala Val Ile 195 200 205 Ser Gly Ser Val Leu Tyr Asn Gln Asp Glu Lys Gly Ser Tyr Ser Leu 210 215 220 Gly Ile Phe Gly Gly Gln Ala Gln Glu Val Ala Gly Ser Ala Glu Val 225 230 235 240 Glu Thr Ala Asn Gly Ile His His Ile Gly Leu Ala Ala Lys Gln 245 250 255 <210> 5 <211> 263 <212> PRT <213> Neisseria meningitidis <400> 5 Cys Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Val Ala Ala Asp 1 5 10 15 Ile Gly Ala Gly Leu Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys 20 25 30 Asp Lys Gly Leu Lys Ser Leu Thr Leu Glu Asp Ser Ile Ser Gln Asn 35 40 45 Gly Thr Leu Thr Leu Ser Ala Gln Gly Ala Glu Arg Thr Phe Lys Ala 50 55 60 Gly Asp Lys Asp Asn Ser Leu Asn Thr Gly Lys Leu Lys Asn Asp Lys 65 70 75 80 Ile Ser Arg Phe Asp Phe Ile Arg Gln Ile Glu Val Asp Gly Gln Leu 85 90 95 Ile Thr Leu Glu Ser Gly Glu Phe Gln Val Tyr Lys Gln Ser His Ser 100 105 110 Ala Leu Thr Ala Leu Gln Thr Glu Gln Val Gln Asp Ser Glu His Ser 115 120 125 Gly Lys Met Val Ala Lys Arg Gln Phe Arg Ile Gly Asp Ile Val Gly 130 135 140 Glu His Thr Ser Phe Gly Lys Leu Pro Lys Asp Val Met Ala Thr Tyr 145 150 155 160 Arg Gly Thr Ala Phe Gly Ser Asp Asp Ala Gly Gly Lys Leu Thr Tyr 165 170 175 Thr Ile Asp Phe Ala Ala Lys Gln Gly His Gly Lys Ile Glu His Leu 180 185 190 Lys Ser Pro Glu Leu Asn Val Asp Leu Ala Ala Ala Asp Ile Lys Pro 195 200 205 Asp Glu Lys His His Ala Val Ile Ser Gly Ser Val Leu Tyr Asn Gln 210 215 220 Ala Glu Lys Gly Ser Tyr Ser Leu Gly Ile Phe Gly Gly Gln Ala Gln 225 230 235 240 Glu Val Ala Gly Ser Ala Glu Val Glu Thr Ala Asn Gly Ile Arg His 245 250 255 Ile Gly Leu Ala Ala Lys Gln 260 <210> 6 <211> 260 <212> PRT <213> Neisseria meningitidis <400> 6 Cys Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Val Thr Ala Asp 1 5 10 15 Ile Gly Thr Gly Leu Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys 20 25 30 Asp Lys Gly Leu Lys Ser Leu Thr Leu Glu Asp Ser Ile Ser Gln Asn 35 40 45 Gly Thr Leu Thr Leu Ser Ala Gln Gly Ala Glu Lys Thr Tyr Gly Asn 50 55 60 Gly Asp Ser Leu Asn Thr Gly Lys Leu Lys Asn Asp Lys Val Ser Arg 65 70 75 80 Phe Asp Phe Ile Arg Gln Ile Glu Val Asp Gly Gln Leu Ile Thr Leu 85 90 95 Glu Ser Gly Glu Phe Gln Val Tyr Lys Gln Ser His Ser Ala Leu Thr 100 105 110 Ala Leu Gln Thr Glu Gln Glu Gln Asp Pro Glu His Ser Glu Lys Met 115 120 125 Val Ala Lys Arg Arg Phe Arg Ile Gly Asp Ile Ala Gly Glu His Thr 130 135 140 Ser Phe Asp Lys Leu Pro Lys Asp Val Met Ala Thr Tyr Arg Gly Thr 145 150 155 160 Ala Phe Gly Ser Asp Asp Ala Gly Gly Lys Leu Thr Tyr Thr Ile Asp 165 170 175 Phe Ala Ala Lys Gln Gly His Gly Lys Ile Glu His Leu Lys Ser Pro 180 185 190 Glu Leu Asn Val Asp Leu Ala Val Ala Tyr Ile Lys Pro Asp Glu Lys 195 200 205 His His Ala Val Ile Ser Gly Ser Val Leu Tyr Asn Gln Asp Glu Lys 210 215 220 Gly Ser Tyr Ser Leu Gly Ile Phe Gly Glu Lys Ala Gln Glu Val Ala 225 230 235 240 Gly Ser Ala Glu Val Glu Thr Ala Asn Gly Ile His His Ile Gly Leu 245 250 255 Ala Ala Lys Gln 260 <210> 7 <211> 254 <212> PRT <213> Neisseria meningitidis <400> 7 Cys Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu 1 5 10 15 Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp Lys Ser Leu Gln 20 25 30 Ser Leu Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu 35 40 45 Ala Ala Gln Gly Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn 50 55 60 Thr Gly Lys Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile Arg 65 70 75 80 Gln Ile Glu Val Asp Gly Gln Leu Ile Thr Leu Glu Ser Gly Glu Phe 85 90 95 Gln Ile Tyr Lys Gln Asp His Ser Ala Val Val Ala Leu Gln Ile Glu 100 105 110 Lys Ile Asn Asn Pro Asp Lys Ile Asp Ser Leu Ile Asn Gln Arg Ser 115 120 125 Phe Leu Val Ser Gly Leu Gly Gly Glu His Thr Ala Phe Asn Gln Leu 130 135 140 Pro Asp Gly Lys Ala Glu Tyr His Gly Lys Ala Phe Ser Ser Asp Asp 145 150 155 160 Ala Gly Gly Lys Leu Thr Tyr Thr Ile Asp Phe Ala Ala Lys Gln Gly 165 170 175 His Gly Lys Ile Glu His Leu Lys Thr Pro Glu Gln Asn Val Glu Leu 180 185 190 Ala Ala Ala Glu Leu Lys Ala Asp Glu Lys Ser His Ala Val Ile Leu 195 200 205 Gly Asp Thr Arg Tyr Gly Ser Glu Glu Lys Gly Thr Tyr His Leu Ala 210 215 220 Leu Phe Gly Asp Arg Ala Gln Glu Ile Ala Gly Ser Ala Thr Val Lys 225 230 235 240 Ile Gly Glu Lys Val His Glu Ile Gly Ile Ala Gly Lys Gln 245 250 <210> 8 <211> 254 <212> PRT <213> Neisseria meningitidis <400> 8 Cys Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu 1 5 10 15 Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp Lys Ser Leu Gln 20 25 30 Ser Leu Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu 35 40 45 Ala Ala Gln Gly Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn 50 55 60 Thr Gly Lys Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile Arg 65 70 75 80 Gln Ile Glu Val Asp Gly Gln Leu Ile Thr Leu Glu Ser Gly Glu Phe 85 90 95 Gln Ile Tyr Lys Gln Asp His Ser Ala Val Val Ala Leu Gln Ile Glu 100 105 110 Lys Ile Asn Asn Pro Asp Lys Ile Asp Ser Leu Ile Asn Gln Arg Ser 115 120 125 Phe Leu Val Ser Gly Leu Gly Gly Glu His Thr Ala Phe Asn Gln Leu 130 135 140 Pro Ser Gly Lys Ala Glu Tyr His Gly Lys Ala Phe Ser Ser Asp Asp 145 150 155 160 Ala Gly Gly Lys Leu Thr Tyr Thr Ile Asp Phe Ala Ala Lys Gln Gly 165 170 175 His Gly Lys Ile Glu His Leu Lys Thr Pro Glu Gln Asn Val Glu Leu 180 185 190 Ala Ser Ala Glu Leu Lys Ala Asp Glu Lys Ser His Ala Val Ile Leu 195 200 205 Gly Asp Thr Arg Tyr Gly Gly Glu Glu Lys Gly Thr Tyr His Leu Ala 210 215 220 Leu Phe Gly Asp Arg Ala Gln Glu Ile Ala Gly Ser Ala Thr Val Lys 225 230 235 240 Ile Arg Glu Lys Val His Glu Ile Gly Ile Ala Gly Lys Gln 245 250 <210> 9 <211> 254 <212> PRT <213> Neisseria meningitidis <400> 9 Cys Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu 1 5 10 15 Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp Lys Ser Leu Gln 20 25 30 Ser Leu Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu 35 40 45 Ala Ala Gln Gly Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn 50 55 60 Thr Gly Lys Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile Arg 65 70 75 80 Gln Ile Glu Val Asp Gly Gln Leu Ile Thr Leu Glu Ser Gly Glu Phe 85 90 95 Gln Ile Tyr Lys Gln Asp His Ser Ala Val Val Ala Leu Gln Ile Glu 100 105 110 Lys Ile Asn Asn Pro Asp Lys Ile Asp Ser Leu Ile Asn Gln Arg Ser 115 120 125 Phe Leu Val Ser Gly Leu Gly Gly Glu His Thr Ala Phe Asn Gln Leu 130 135 140 Pro Ser Gly Lys Ala Glu Tyr His Gly Lys Ala Phe Ser Ser Asp Asp 145 150 155 160 Pro Asn Gly Arg Leu His Tyr Ser Ile Asp Phe Thr Lys Lys Gln Gly 165 170 175 Tyr Gly Arg Ile Glu His Leu Lys Thr Pro Glu Gln Asn Val Glu Leu 180 185 190 Ala Ser Ala Glu Leu Lys Ala Asp Glu Lys Ser His Ala Val Ile Leu 195 200 205 Gly Asp Thr Arg Tyr Gly Gly Glu Glu Lys Gly Thr Tyr His Leu Ala 210 215 220 Leu Phe Gly Asp Arg Ala Gln Glu Ile Ala Gly Ser Ala Thr Val Lys 225 230 235 240 Ile Arg Glu Lys Val His Glu Ile Gly Ile Ala Gly Lys Gln 245 250 <210> 10 <211> 254 <212> PRT <213> Neisseria meningitidis <400> 10 Cys Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu 1 5 10 15 Ala Asp Ala Leu Thr Thr Pro Leu Asp His Lys Asp Lys Ser Leu Gln 20 25 30 Ser Leu Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu 35 40 45 Ala Ala Gln Gly Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn 50 55 60 Thr Gly Lys Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile Arg 65 70 75 80 Gln Ile Glu Val Asp Gly Gln Thr Ile Thr Leu Ala Ser Gly Glu Phe 85 90 95 Gln Ile Tyr Lys Gln Asn His Ser Ala Val Val Ala Leu Gln Ile Glu 100 105 110 Lys Ile Asn Asn Pro Asp Lys Ile Asp Ser Leu Ile Asn Gln Arg Ser 115 120 125 Phe Leu Val Ser Gly Leu Gly Gly Glu His Thr Ala Phe Asn Gln Leu 130 135 140 Pro Asp Gly Lys Ala Glu Tyr His Gly Lys Ala Phe Ser Ser Asp Asp 145 150 155 160 Pro Asn Gly Arg Leu His Tyr Ser Ile Asp Phe Thr Lys Lys Gln Gly 165 170 175 Tyr Gly Arg Ile Glu His Leu Lys Thr Pro Glu Gln Asn Val Glu Leu 180 185 190 Ala Ser Ala Glu Leu Lys Ala Asp Glu Lys Ser His Ala Val Ile Leu 195 200 205 Gly Asp Thr Arg Tyr Gly Gly Glu Glu Lys Gly Thr Tyr His Leu Ala 210 215 220 Leu Phe Gly Asp Arg Ala Gln Glu Ile Ala Gly Ser Ala Thr Val Lys 225 230 235 240 Ile Arg Glu Lys Val His Glu Ile Gly Ile Ala Gly Lys Gln 245 250 <210> 11 <211> 261 <212> PRT <213> Neisseria meningitidis <400> 11 Cys Ser Ser Gly Ser Gly Ser Gly Gly Gly Gly Val Ala Ala Asp Ile 1 5 10 15 Gly Thr Gly Leu Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp 20 25 30 Lys Gly Leu Lys Ser Leu Thr Leu Glu Asp Ser Ile Ser Gln Asn Gly 35 40 45 Thr Leu Thr Leu Ser Ala Gln Gly Ala Glu Lys Thr Phe Lys Val Gly 50 55 60 Asp Lys Asp Asn Ser Leu Asn Thr Gly Lys Leu Lys Asn Asp Lys Ile 65 70 75 80 Ser Arg Phe Asp Phe Val Gln Lys Ile Glu Val Asp Gly Gln Thr Ile 85 90 95 Thr Leu Ala Ser Gly Glu Phe Gln Ile Tyr Lys Gln Asp His Ser Ala 100 105 110 Val Val Ala Leu Gln Ile Glu Lys Ile Asn Asn Pro Asp Lys Ile Asp 115 120 125 Ser Leu Ile Asn Gln Arg Ser Phe Leu Val Ser Gly Leu Gly Gly Glu 130 135 140 His Thr Ala Phe Asn Gln Leu Pro Ser Gly Lys Ala Glu Tyr His Gly 145 150 155 160 Lys Ala Phe Ser Ser Asp Asp Ala Gly Gly Lys Leu Thr Tyr Thr Ile 165 170 175 Asp Phe Ala Ala Lys Gln Gly His Gly Lys Ile Glu His Leu Lys Thr 180 185 190 Pro Glu Gln Asn Val Glu Leu Ala Ser Ala Glu Leu Lys Ala Asp Glu 195 200 205 Lys Ser His Ala Val Ile Leu Gly Asp Thr Arg Tyr Gly Ser Glu Glu 210 215 220 Lys Gly Thr Tyr His Leu Ala Leu Phe Gly Asp Arg Ala Gln Glu Ile 225 230 235 240 Ala Gly Ser Ala Thr Val Lys Ile Arg Glu Lys Val His Glu Ile Gly 245 250 255 Ile Ala Gly Lys Gln 260 <210> 12 <211> 257 <212> PRT <213> Neisseria meningitidis <400> 12 Cys Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu 1 5 10 15 Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp Lys Gly Leu Lys 20 25 30 Ser Leu Thr Leu Glu Asp Ser Ile Ser Gln Asn Gly Thr Leu Thr Leu 35 40 45 Ser Ala Gln Gly Ala Glu Lys Thr Phe Lys Val Gly Asp Lys Asp Asn 50 55 60 Ser Leu Asn Thr Gly Lys Leu Lys Asn Asp Lys Ile Ser Arg Phe Asp 65 70 75 80 Phe Val Gln Lys Ile Glu Val Asp Gly Gln Thr Ile Thr Leu Ala Ser 85 90 95 Gly Glu Phe Gln Ile Tyr Lys Gln Asn His Ser Ala Val Val Ala Leu 100 105 110 Gln Ile Glu Lys Ile Asn Asn Pro Asp Lys Ile Asp Ser Leu Ile Asn 115 120 125 Gln Arg Ser Phe Leu Val Ser Gly Leu Gly Gly Glu His Thr Ala Phe 130 135 140 Asn Gln Leu Pro Gly Gly Lys Ala Glu Tyr His Gly Lys Ala Phe Ser 145 150 155 160 Ser Asp Asp Ala Gly Gly Lys Leu Thr Tyr Thr Ile Asp Phe Ala Ala 165 170 175 Lys Gln Gly His Gly Lys Ile Glu His Leu Lys Thr Pro Glu Gln Asn 180 185 190 Val Glu Leu Ala Ala Ala Glu Leu Lys Ala Asp Glu Lys Ser His Ala 195 200 205 Val Ile Leu Gly Asp Thr Arg Tyr Gly Ser Glu Glu Lys Gly Thr Tyr 210 215 220 His Leu Ala Leu Phe Gly Asp Arg Ala Gln Glu Ile Ala Gly Ser Ala 225 230 235 240 Thr Val Lys Ile Gly Glu Lys Val His Glu Ile Ser Ile Ala Gly Lys 245 250 255 Gln <210> 13 <211> 285 <212> PRT <213> Neisseria meningitidis <400> 13 Met Thr Arg Ser Lys Pro Val Asn Arg Thr Thr Phe Cys Cys Leu Ser 1 5 10 15 Leu Thr Ala Gly Pro Asp Ser Asp Arg Leu Gln Gln Arg Arg Gly Gly 20 25 30 Gly Gly Gly Val Ala Ala Asp Ile Gly Thr Gly Leu Ala Asp Ala Leu 35 40 45 Thr Ala Pro Leu Asp His Lys Asp Lys Gly Leu Lys Ser Leu Thr Leu 50 55 60 Glu Ala Ser Ile Pro Gln Asn Gly Thr Leu Thr Leu Ser Ala Gln Gly 65 70 75 80 Ala Glu Lys Thr Phe Lys Ala Gly Gly Lys Asp Asn Ser Leu Asn Thr 85 90 95 Gly Lys Leu Lys Asn Asp Lys Ile Ser Arg Phe Asp Phe Val Gln Lys 100 105 110 Ile Glu Val Asp Gly Gln Thr Ile Thr Leu Ala Ser Gly Glu Phe Gln 115 120 125 Ile Tyr Lys Gln Asp His Ser Ala Val Val Ala Leu Arg Ile Glu Lys 130 135 140 Ile Asn Asn Pro Asp Lys Ile Asp Ser Leu Ile Asn Gln Arg Ser Phe 145 150 155 160 Leu Val Ser Asp Leu Gly Gly Glu His Thr Ala Phe Asn Gln Leu Pro 165 170 175 Asp Gly Lys Ala Glu Tyr His Gly Lys Ala Phe Ser Ser Asp Asp Ala 180 185 190 Asp Gly Lys Leu Thr Tyr Thr Ile Asp Phe Ala Ala Lys Gln Gly His 195 200 205 Gly Lys Ile Glu His Leu Lys Thr Pro Glu Gln Asn Val Glu Leu Ala 210 215 220 Ser Ala Glu Leu Lys Ala Asp Glu Lys Ser His Ala Val Ile Leu Gly 225 230 235 240 Asp Thr Arg Tyr Gly Gly Glu Glu Lys Gly Thr Tyr Arg Leu Ala Leu 245 250 255 Phe Gly Asp Arg Ala Gln Glu Ile Ala Gly Ser Ala Thr Val Lys Ile 260 265 270 Gly Glu Lys Val His Glu Ile Gly Ile Ala Asp Lys Gln 275 280 285 <210> 14 <211> 256 <212> PRT <213> Neisseria meningitidis <400> 14 Cys Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu 1 5 10 15 Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp Lys Gly Leu Gln 20 25 30 Ser Leu Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu 35 40 45 Ala Ala Pro Gln Gly Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu 50 55 60 Asn Thr Gly Lys Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile 65 70 75 80 Arg Gln Ile Glu Val Asp Gly Gln Leu Ile Thr Leu Glu Ser Gly Glu 85 90 95 Phe Gln Val Tyr Lys Gln Ser His Ser Ala Leu Thr Ala Phe Gln Thr 100 105 110 Glu Gln Ile Gln Asp Ser Glu His Ser Gly Lys Met Val Ala Lys Arg 115 120 125 Gln Phe Arg Ile Gly Asp Ile Ala Gly Glu His Thr Ser Phe Asp Lys 130 135 140 Leu Pro Glu Gly Gly Arg Ala Thr Tyr Arg Gly Thr Ala Phe Gly Ser 145 150 155 160 Asp Asp Ala Gly Gly Lys Leu Thr Tyr Thr Ile Asp Phe Ala Ala Lys 165 170 175 Gln Gly Asn Gly Lys Ile Glu His Leu Lys Ser Pro Glu Leu Asn Val 180 185 190 Asp Leu Ala Ala Ala Asp Ile Lys Pro Asp Gly Lys Arg His Ala Val 195 200 205 Ile Ser Gly Ser Val Leu Tyr Asn Gln Ala Glu Lys Gly Ser Tyr Ser 210 215 220 Leu Gly Ile Phe Gly Gly Lys Ala Gln Glu Val Ala Gly Ser Ala Glu 225 230 235 240 Val Lys Thr Val Asn Gly Ile Arg His Ile Gly Leu Ala Ala Lys Gln 245 250 255 <210> 15 <211> 6 <212> PRT <213> Neisseria meningitidis <400> 15 Glu Val Asp Gly Gln Leu 1 5 <210> 16 <211> 8 <212> PRT <213> Neisseria meningitidis <400> 16 Lys Pro Asp Gly Lys Arg His Ala 1 5 <210> 17 <211> 7 <212> PRT <213> Neisseria meningitidis <400> 17 Phe Gly Gly Lys Ala Gln Glu 1 5 <210> 18 <211> 6 <212> PRT <213> Neisseria meningitidis <400> 18 Ala Ala Gln Gly Ala Glu 1 5 <210> 19 <211> 10 <212> PRT <213> Neisseria meningitidis <400> 19 Ile Gln Asp Ser Glu His Ser Gly Lys Met 1 5 10 <210> 20 <211> 6 <212> PRT <213> Neisseria meningitidis <400> 20 Lys Thr Val Asn Gly Ile 1 5 <210> twenty one <211> 7 <212> PRT <213> Neisseria meningitidis <220> <221> XA <222> (1) <223> Exogenous ring insertion <400> twenty one Xaa Ala Ala Gln Gly Ala Glu 1 5 <210> twenty two <211> 7 <212> PRT <213> Artificial sequence <220> <223> Modified peptides <220> <221> XA <222> (2)..(2) <223> Exogenous ring insertion <400> twenty two Ala Xaa Ala Gln Gly Ala Glu 1 5 <210> twenty three <211> 7 <212> PRT <213> Artificial sequence <220> <223> Modified peptides <220> <221> XA <222> (3) <223> Exogenous ring insertion <400> twenty three Ala Ala Xaa Gln Gly Ala Glu 1 5 <210> twenty four <211> 7 <212> PRT <213> Artificial sequence <220> <223> Modified peptides <220> <221> XA <222> (4) <223> Exogenous ring insertion <400> twenty four Ala Ala Gln Xaa Gly Ala Glu 1 5 <210> 25 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Modified peptides <220> <221> XA <222> (5) <223> Exogenous ring insertion <400> 25 Ala Ala Gln Gly Xaa Ala Glu 1 5 <210> 26 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Modified peptides <220> <221> XA <222> (6) <223> Exogenous ring insertion <400> 26 Ala Ala Gln Gly Ala Xaa Glu 1 5 <210> 27 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Modified peptides <220> <221> XA <222> (7) <223> Exogenous ring insertion <400> 27 Ala Ala Gln Gly Ala Glu Xaa 1 5 <210> 28 <211> 6 <212> PRT <213> Artificial sequence <220> <223> Modified peptides <220> <221> XA <222> (2) <223> Exogenous ring insertion <400> 28 Ala Xaa Gln Gly Ala Glu 1 5 <210> 29 <211> 6 <212> PRT <213> Artificial sequence <220> <223> Modified peptides <220> <221> XA <222> (3) <223> Exogenous ring insertion <400> 29 Ala Ala Xaa Gly Ala Glu 1 5 <210> 30 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Modified peptides <220> <221> XA <222> (3) <223> Exogenous ring insertion <400> 30 Ala Ala Xaa Ala Glu 1 5 <210> 31 <211> 4 <212> PRT <213> Artificial sequence <220> <223> Modified peptides <220> <221> misc_features <222> (3) <223> Xaa can be any naturally occurring amino acid <400> 31 Ala Ala Xaa Glu 1 <210> 32 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Modified peptides <220> <221> XA <222> (2) <223> Exogenous ring insertion <400> 32 Ala Xaa Gly Ala Glu 1 5 <210> 33 <211> 4 <212> PRT <213> Artificial sequence <220> <223> Modified peptides <220> <221> XA <222> (2)..(2) <223> Exogenous ring insertion <400> 33 Ala Xaa Ala Glu 1 <210> 34 <211> 4 <212> PRT <213> Artificial sequence <220> <223> Modified peptides <220> <221> XA <222> (3) <223> Exogenous ring insertion <400> 34 Ala Ala Xaa Glu 1 <210> 35 <211> 6 <212> PRT <213> Artificial sequence <220> <223> Modified peptides <220> <221> XA <222> (1) <223> Exogenous ring insertion <400> 35 Xaa Ala Gln Gly Ala Glu 1 5 <210> 36 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Modified peptides <220> <221> XA <222> (1) <223> Exogenous ring insertion <400> 36 Xaa Gln Gly Ala Glu 1 5 <210> 37 <211> 4 <212> PRT <213> Artificial sequence <220> <223> Modified peptides <220> <221> XA <222> (1) <223> Exogenous ring insertion <400> 37 Xaa Gly Ala Glu 1 <210> 38 <211> 6 <212> PRT <213> Artificial sequence <220> <223> Modified peptides <220> <221> XA <222> (4) <223> Exogenous ring insertion <400> 38 Ala Ala Gln Xaa Ala Glu 1 5 <210> 39 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Modified peptides <220> <221> XA <222> (4) <223> Exogenous ring insertion <400> 39 Ala Ala Gln Xaa Glu 1 5 <210> 40 <211> 4 <212> PRT <213> Artificial sequence <220> <223> Modified peptides <220> <221> XA <222> (4) <223> Exogenous ring insertion <400> 40 Ala Ala Gln Xaa 1 <210> 41 <211> 6 <212> PRT <213> Artificial sequence <220> <223> Modified peptides <220> <221> XA <222> (5) <223> Exogenous ring insertion <400> 41 Ala Ala Gln Gly Xaa Glu 1 5 <210> 42 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Modified peptides <220> <221> XA <222> (5) <223> Exogenous ring insertion <400> 42 Ala Ala Gln Gly Xaa 1 5 <210> 43 <211> 6 <212> PRT <213> Artificial sequence <220> <223> Modified peptides <220> <221> XA <222> (6)..(6) <223> Exogenous loop insertion <400> 43 Ala Ala Gln Gly Ala Xaa 1 5 <210> 44 <211> 255 <212> PRT <213> Neisseria meningitidis <400> 44 Cys Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu 1 5 10 15 Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp Lys Gly Leu Gln 20 25 30 Ser Leu Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu 35 40 45 Ala Ala Gln Gly Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn 50 55 60 Thr Gly Lys Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile Arg 65 70 75 80 Gln Ile Glu Val Asp Gly Gln Leu Ile Thr Leu Glu Ser Gly Glu Phe 85 90 95 Gln Val Tyr Lys Gln Ser His Ser Ala Leu Thr Ala Phe Gln Thr Glu 100 105 110 Gln Ile Gln Asp Ser Glu His Ser Gly Lys Met Val Ala Lys Arg Gln 115 120 125 Phe Arg Ile Gly Asp Ile Ala Gly Glu His Thr Ser Phe Asp Lys Leu 130 135 140 Pro Glu Gly Gly Arg Ala Thr Tyr Arg Gly Thr Ala Phe Gly Ser Asp 145 150 155 160 Asp Ala Gly Gly Lys Leu Thr Tyr Thr Ile Asp Phe Ala Ala Lys Gln 165 170 175 Gly Asn Gly Lys Ile Glu His Leu Lys Ser Pro Glu Leu Asn Val Asp 180 185 190 Leu Ala Ala Ala Asp Ile Lys Pro Asp Gly Lys Arg His Ala Val Ile 195 200 205 Ser Gly Ser Val Leu Tyr Asn Gln Ala Glu Lys Gly Ser Tyr Ser Leu 210 215 220 Gly Ile Phe Gly Gly Lys Ala Gln Glu Val Ala Gly Ser Ala Glu Val 225 230 235 240 Lys Thr Val Asn Gly Ile Arg His Ile Gly Leu Ala Ala Lys Gln 245 250 255 <210> 45 <211> 13 <212> PRT <213> Neisseria meningitidis <400> 45 Tyr Val Ala Val Glu Asn Gly Val Ala Lys Lys Val Ala 1 5 10 <210> 46 <211> 15 <212> PRT <213> Neisseria meningitidis <400> 46 His Phe Val Gln Gln Thr Pro Lys Ser Gln Pro Thr Leu Val Pro 1 5 10 15 <210> 47 <211> 15 <212> PRT <213> Neisseria meningitidis <400> 47 His Phe Val Gln Gln Thr Pro Gln Ser Gln Pro Thr Leu Val Pro 1 5 10 15 <210> 48 <211> 14 <212> PRT <213> Neisseria meningitidis <400> 48 Thr Leu Ala Asn Gly Ala Asn Asn Thr Ile Ile Arg Val Pro 1 5 10 <210> 49 <211> 14 <212> PRT <213> Neisseria meningitidis <400> 49 Thr Leu Ala Lys Gly Ala Asn Asn Thr Ile Ile Arg Val Pro 1 5 10 <210> 50 <211> 13 <212> PRT <213> Neisseria meningitidis <400> 50 His Val Val Val Asn Asn Lys Val Ala Thr His Val Pro 1 5 10 <210> 51 <211> 10 <212> PRT <213> Neisseria meningitidis <400> 51 Tyr Val Asp Glu Gln Ser Lys Tyr His Ala 1 5 10 <210> 52 <211> 15 <212> PRT <213> Neisseria meningitidis <400> 52 His Phe Val Gln Asn Lys Gln Asn Gln Arg Pro Thr Leu Val Pro 1 5 10 15 <210> 53 <211> 15 <212> PRT <213> Neisseria meningitidis <400> 53 His Phe Val Gln Asn Lys Gln Asn Gln Pro Pro Thr Leu Val Pro 1 5 10 15 <210> 54 <211> 15 <212> PRT <213> Neisseria meningitidis <400> 54 His Phe Val Gln Asp Lys Lys Gly Gln Pro Pro Thr Leu Val Pro 1 5 10 15 <210> 55 <211> 18 <212> PRT <213> Neisseria meningitidis <400> 55 His Phe Val Gln Asn Lys Gln Asn Gln Gln Asn Gln Pro Pro Thr Leu 1 5 10 15 Val Pro <210> 56 <211> 18 <212> PRT <213> Neisseria meningitidis <400> 56 His Phe Val Gln Asn Lys Gln Asn Lys Gln Asn Gln Pro Pro Thr Leu 1 5 10 15 Val Pro <210> 57 <211> 15 <212> PRT <213> Neisseria meningitidis <400> 57 His Phe Val Gln Asn Lys Gln Asn Lys Pro Pro Thr Leu Val Pro 1 5 10 15 <210> 58 <211> 18 <212> PRT <213> Neisseria meningitidis <400> 58 Tyr Trp Thr Thr Val Asn Thr Gly Ser Ala Thr Thr Thr Thr Thr Phe 1 5 10 15 Val Pro <210> 59 <211> 17 <212> PRT <213> Neisseria meningitidis <400> 59 Tyr Trp Thr Thr Val Asn Thr Gly Ser Ala Thr Thr Thr Thr Phe Val 1 5 10 15 Pro <210> 60 <211> 16 <212> PRT <213> Neisseria meningitidis <400> 60 Tyr Trp Thr Thr Val Asn Thr Gly Ser Ala Thr Thr Thr Phe Val Pro 1 5 10 15 <210> 61 <211> 17 <212> PRT <213> Neisseria meningitidis <400> 61 Tyr Tyr Thr Thr Val Thr Gln Gly Ser Ala Thr Thr Thr Thr Phe Val 1 5 10 15 Pro <210> 62 <211> 10 <212> PRT <213> Neisseria meningitidis <400> 62 Tyr Val Asp Glu Lys Lys Met Val His Ala 1 5 10 <210> 63 <211> 10 <212> PRT <213> Neisseria meningitidis <400> 63 Tyr Val Asp Glu Lys Gln Val Ser His Ala 1 5 10 <210> 64 <211> 10 <212> PRT <213> Neisseria meningitidis <400> 64 Tyr Val Asp Glu Lys Lys Val Val His Ala 1 5 10 <210> 65 <211> 13 <212> PRT <213> Neisseria meningitidis <400> 65 His Tyr Thr Arg Gln Asn Asn Ala Asp Val Phe Val Pro 1 5 10 <210> 66 <211> 13 <212> PRT <213> Neisseria meningitidis <400> 66 His Tyr Thr Arg Gln Asn Asn Thr Asp Val Phe Val Pro 1 5 10 <210> 67 <211> 13 <212> PRT <213> Neisseria meningitidis <400> 67 His Tyr Thr Arg Gln Asn Asn Ile Asp Val Phe Val Pro 1 5 10 <210> 68 <211> 14 <212> PRT <213> Neisseria meningitidis <400> 68 Tyr Tyr Thr Lys Asp Thr Asn Asn Asn Leu Thr Leu Val Pro 1 5 10 <210> 69 <211> 14 <212> PRT <213> Neisseria meningitidis <400> 69 Tyr Tyr Thr Lys Asp Lys Asn Asp Asn Leu Thr Leu Val Pro 1 5 10 <210> 70 <211> 14 <212> PRT <213> Neisseria meningitidis <400> 70 Tyr Tyr Thr Lys Asp Lys Asn Asp Lys Leu Thr Leu Val Pro 1 5 10 <210> 71 <211> 13 <212> PRT <213> Neisseria meningitidis <400> 71 Tyr Tyr Thr Asn Thr Asn Asn Asn Leu Thr Leu Val Pro 1 5 10 <210> 72 <211> 17 <212> PRT <213> Neisseria meningitidis <400> 72 His Trp Asn Thr Val Tyr Asn Thr Asn Gly Thr Thr Thr Thr Phe Val 1 5 10 15 Pro <210> 73 <211> 18 <212> PRT <213> Neisseria meningitidis <400> 73 His Trp Asn Thr Val Tyr Asn Thr Asn Gly Thr Thr Thr Thr Thr Phe 1 5 10 15 Val Pro <210> 74 <211> 14 <212> PRT <213> Neisseria meningitidis <400> 74 Thr Tyr Thr Val Asp Ser Ser Gly Val Val Thr Pro Val Pro 1 5 10 <210> 75 <211> 14 <212> PRT <213> Neisseria meningitidis <400> 75 His Phe Val Ala Asp Ser Gln Gly Lys Ile Thr Arg Val Pro 1 5 10 <210> 76 <211> 17 <212> PRT <213> Neisseria meningitidis <400> 76 Tyr Tyr Tyr Thr Thr Ala Thr Asn Ser Ser Thr Ser Thr Thr Phe Val 1 5 10 15 Pro <210> 77 <211> 17 <212> PRT <213> Neisseria meningitidis <400> 77 His Tyr Thr Thr Val Tyr Asn Ala Thr Thr Thr Thr Thr Thr Phe Val 1 5 10 15 Pro <210> 78 <211> 18 <212> PRT <213> Neisseria meningitidis <400> 78 His Tyr Thr Thr Val Tyr Asn Ala Thr Thr Thr Thr Thr Thr Thr Thr Phe 1 5 10 15 Val Pro <210> 79 <211> 11 <212> PRT <213> Neisseria meningitidis <400> 79 Tyr Val Asp Asp Gln Gly Lys Val Lys Gly Pro 1 5 10 <210> 80 <211> 255 <212> PRT <213> Neisseria meningitidis <400> 80 Cys Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu 1 5 10 15 Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp Lys Gly Leu Gln 20 25 30 Ser Leu Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu 35 40 45 Ala Ala Gln Gly Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn 50 55 60 Thr Gly Lys Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile Arg 65 70 75 80 Gln Ile Glu Val Asp Gly Gln Leu Ile Thr Leu Glu Ser Gly Glu Phe 85 90 95 Gln Val Tyr Lys Gln Ser His Ser Ala Leu Thr Ala Phe Gln Thr Glu 100 105 110 Gln Ile Gln Asp Ser Glu His Ser Gly Lys Met Val Ala Lys Arg Gln 115 120 125 Phe Arg Ile Gly Asp Ile Ala Gly Glu His Thr Ser Phe Asp Lys Leu 130 135 140 Pro Glu Gly Gly Arg Ala Thr Tyr Arg Gly Thr Ala Phe Gly Ser Asp 145 150 155 160 Asp Ala Gly Gly Lys Leu Thr Tyr Thr Ile Asp Phe Ala Ala Lys Gln 165 170 175 Gly Asn Gly Lys Ile Glu His Leu Lys Ser Pro Glu Leu Asn Val Asp 180 185 190 Leu Ala Ala Ala Asp Ile Lys Pro Asp Gly Lys Arg His Ala Val Ile 195 200 205 Ser Gly Ser Val Leu Tyr Asn Gln Ala Glu Lys Gly Ser Tyr Ser Leu 210 215 220 Gly Ile Phe Gly Gly Lys Ala Gln Glu Val Ala Gly Ser Ala Glu Val 225 230 235 240 Lys Thr Val Asn Gly Ile Arg His Ile Gly Leu Ala Ala Lys Gln 245 250 255 <210> 81 <211> 255 <212> PRT <213> Neisseria meningitidis <400> 81 Cys Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu 1 5 10 15 Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp Lys Ser Leu Gln 20 25 30 Ser Leu Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu 35 40 45 Ala Ala Gln Gly Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn 50 55 60 Thr Gly Lys Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile Arg 65 70 75 80 Gln Ile Glu Val Asp Gly Gln Leu Ile Thr Leu Glu Ser Gly Glu Phe 85 90 95 Gln Val Tyr Lys Gln Ser His Ser Ala Leu Thr Ala Leu Gln Thr Glu 100 105 110 Gln Val Gln Asp Ser Glu His Ser Gly Lys Met Val Ala Lys Arg Gln 115 120 125 Phe Arg Ile Gly Asp Ile Ala Gly Glu His Thr Ser Phe Asp Lys Leu 130 135 140 Pro Glu Gly Gly Arg Ala Thr Tyr Arg Gly Thr Ala Phe Gly Ser Asp 145 150 155 160 Asp Ala Ser Gly Lys Leu Thr Tyr Thr Ile Asp Phe Ala Ala Lys Gln 165 170 175 Gly His Gly Lys Ile Glu His Leu Lys Ser Pro Glu Leu Asn Val Asp 180 185 190 Leu Ala Ala Ser Asp Ile Lys Pro Asp Lys Lys Arg His Ala Val Ile 195 200 205 Ser Gly Ser Val Leu Tyr Asn Gln Ala Glu Lys Gly Ser Tyr Ser Leu 210 215 220 Gly Ile Phe Gly Gly Gln Ala Gln Glu Val Ala Gly Ser Ala Glu Val 225 230 235 240 Glu Thr Ala Asn Gly Ile Arg His Ile Gly Leu Ala Ala Lys Gln 245 250 255 <210> 82 <211> 255 <212> PRT <213> Neisseria meningitidis <400> 82 Cys Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu 1 5 10 15 Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp Lys Gly Leu Gln 20 25 30 Ser Leu Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu 35 40 45 Ala Ala Gln Gly Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn 50 55 60 Thr Gly Lys Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile Arg 65 70 75 80 Gln Ile Glu Val Asp Gly Lys Leu Ile Thr Leu Glu Ser Gly Glu Phe 85 90 95 Gln Val Tyr Lys Gln Ser His Ser Ala Leu Thr Ala Leu Gln Thr Glu 100 105 110 Gln Val Gln Asp Ser Glu Asp Ser Gly Lys Met Val Ala Lys Arg Gln 115 120 125 Phe Arg Ile Gly Asp Ile Ala Gly Glu His Thr Ser Phe Asp Lys Leu 130 135 140 Pro Lys Gly Gly Ser Ala Thr Tyr Arg Gly Thr Ala Phe Gly Ser Asp 145 150 155 160 Asp Ala Gly Gly Lys Leu Thr Tyr Thr Ile Asp Phe Ala Ala Lys Gln 165 170 175 Gly His Gly Lys Ile Glu His Leu Lys Ser Pro Glu Leu Asn Val Glu 180 185 190 Leu Ala Thr Ala Tyr Ile Lys Pro Asp Glu Lys Arg His Ala Val Ile 195 200 205 Ser Gly Ser Val Leu Tyr Asn Gln Asp Glu Lys Gly Ser Tyr Ser Leu 210 215 220 Gly Ile Phe Gly Gly Gln Ala Gln Glu Val Ala Gly Ser Ala Glu Val 225 230 235 240 Glu Thr Ala Asn Gly Ile His His Ile Gly Leu Ala Ala Lys Gln 245 250 255 <210> 83 <211> 255 <212> PRT <213> Neisseria meningitidis <400> 83 Cys Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu 1 5 10 15 Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp Lys Ser Leu Gln 20 25 30 Ser Leu Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu 35 40 45 Ala Ala Gln Gly Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn 50 55 60 Thr Gly Lys Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile Arg 65 70 75 80 Gln Ile Glu Val Asp Gly Gln Leu Ile Thr Leu Glu Ser Gly Glu Phe 85 90 95 Gln Val Tyr Lys Gln Ser His Ser Ala Leu Thr Ala Leu Gln Thr Glu 100 105 110 Gln Glu Gln Asp Pro Glu His Ser Gly Lys Met Val Ala Lys Arg Arg 115 120 125 Phe Lys Ile Gly Asp Ile Ala Gly Glu His Thr Ser Phe Asp Lys Leu 130 135 140 Pro Lys Asp Val Met Ala Thr Tyr Arg Gly Thr Ala Phe Gly Ser Asp 145 150 155 160 Asp Ala Gly Gly Lys Leu Thr Tyr Thr Ile Asp Phe Ala Ala Lys Gln 165 170 175 Gly His Gly Lys Ile Glu His Leu Lys Ser Pro Glu Leu Asn Val Glu 180 185 190 Leu Ala Thr Ala Tyr Ile Lys Pro Asp Glu Lys His His Ala Val Ile 195 200 205 Ser Gly Ser Val Leu Tyr Asn Gln Asp Glu Lys Gly Ser Tyr Ser Leu 210 215 220 Gly Ile Phe Gly Gly Gln Ala Gln Glu Val Ala Gly Ser Ala Glu Val 225 230 235 240 Glu Thr Ala Asn Gly Ile His His Ile Gly Leu Ala Ala Lys Gln 245 250 255 <210> 84 <211> 263 <212> PRT <213> Neisseria meningitidis <400> 84 Cys Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Val Ala Ala Asp 1 5 10 15 Ile Gly Ala Gly Leu Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys 20 25 30 Asp Lys Gly Leu Lys Ser Leu Thr Leu Glu Asp Ser Ile Ser Gln Asn 35 40 45 Gly Thr Leu Thr Leu Ser Ala Gln Gly Ala Glu Arg Thr Phe Lys Ala 50 55 60 Gly Asp Lys Asp Asn Ser Leu Asn Thr Gly Lys Leu Lys Asn Asp Lys 65 70 75 80 Ile Ser Arg Phe Asp Phe Ile Arg Gln Ile Glu Val Asp Gly Gln Leu 85 90 95 Ile Thr Leu Glu Ser Gly Glu Phe Gln Val Tyr Lys Gln Ser His Ser 100 105 110 Ala Leu Thr Ala Leu Gln Thr Glu Gln Val Gln Asp Ser Glu His Ser 115 120 125 Gly Lys Met Val Ala Lys Arg Gln Phe Arg Ile Gly Asp Ile Val Gly 130 135 140 Glu His Thr Ser Phe Gly Lys Leu Pro Lys Asp Val Met Ala Thr Tyr 145 150 155 160 Arg Gly Thr Ala Phe Gly Ser Asp Asp Ala Gly Gly Lys Leu Thr Tyr 165 170 175 Thr Ile Asp Phe Ala Ala Lys Gln Gly His Gly Lys Ile Glu His Leu 180 185 190 Lys Ser Pro Glu Leu Asn Val Asp Leu Ala Ala Ala Asp Ile Lys Pro 195 200 205 Asp Glu Lys His His Ala Val Ile Ser Gly Ser Val Leu Tyr Asn Gln 210 215 220 Ala Glu Lys Gly Ser Tyr Ser Leu Gly Ile Phe Gly Gly Gln Ala Gln 225 230 235 240 Glu Val Ala Gly Ser Ala Glu Val Glu Thr Ala Asn Gly Ile Arg His 245 250 255 Ile Gly Leu Ala Ala Lys Gln 260 <210> 85 <211> 260 <212> PRT <213> Neisseria meningitidis <400> 85 Cys Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Val Thr Ala Asp 1 5 10 15 Ile Gly Thr Gly Leu Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys 20 25 30 Asp Lys Gly Leu Lys Ser Leu Thr Leu Glu Asp Ser Ile Ser Gln Asn 35 40 45 Gly Thr Leu Thr Leu Ser Ala Gln Gly Ala Glu Lys Thr Tyr Gly Asn 50 55 60 Gly Asp Ser Leu Asn Thr Gly Lys Leu Lys Asn Asp Lys Val Ser Arg 65 70 75 80 Phe Asp Phe Ile Arg Gln Ile Glu Val Asp Gly Gln Leu Ile Thr Leu 85 90 95 Glu Ser Gly Glu Phe Gln Val Tyr Lys Gln Ser His Ser Ala Leu Thr 100 105 110 Ala Leu Gln Thr Glu Gln Glu Gln Asp Pro Glu His Ser Glu Lys Met 115 120 125 Val Ala Lys Arg Arg Phe Arg Ile Gly Asp Ile Ala Gly Glu His Thr 130 135 140 Ser Phe Asp Lys Leu Pro Lys Asp Val Met Ala Thr Tyr Arg Gly Thr 145 150 155 160 Ala Phe Gly Ser Asp Asp Ala Gly Gly Lys Leu Thr Tyr Thr Ile Asp 165 170 175 Phe Ala Ala Lys Gln Gly His Gly Lys Ile Glu His Leu Lys Ser Pro 180 185 190 Glu Leu Asn Val Asp Leu Ala Val Ala Tyr Ile Lys Pro Asp Glu Lys 195 200 205 His His Ala Val Ile Ser Gly Ser Val Leu Tyr Asn Gln Asp Glu Lys 210 215 220 Gly Ser Tyr Ser Leu Gly Ile Phe Gly Glu Lys Ala Gln Glu Val Ala 225 230 235 240 Gly Ser Ala Glu Val Glu Thr Ala Asn Gly Ile His His Ile Gly Leu 245 250 255 Ala Ala Lys Gln 260 <210> 86 <211> 254 <212> PRT <213> Neisseria meningitidis <400> 86 Cys Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu 1 5 10 15 Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp Lys Ser Leu Gln 20 25 30 Ser Leu Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu 35 40 45 Ala Ala Gln Gly Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn 50 55 60 Thr Gly Lys Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile Arg 65 70 75 80 Gln Ile Glu Val Asp Gly Gln Leu Ile Thr Leu Glu Ser Gly Glu Phe 85 90 95 Gln Ile Tyr Lys Gln Asp His Ser Ala Val Val Ala Leu Gln Ile Glu 100 105 110 Lys Ile Asn Asn Pro Asp Lys Ile Asp Ser Leu Ile Asn Gln Arg Ser 115 120 125 Phe Leu Val Ser Gly Leu Gly Gly Glu His Thr Ala Phe Asn Gln Leu 130 135 140 Pro Asp Gly Lys Ala Glu Tyr His Gly Lys Ala Phe Ser Ser Asp Asp 145 150 155 160 Ala Gly Gly Lys Leu Thr Tyr Thr Ile Asp Phe Ala Ala Lys Gln Gly 165 170 175 His Gly Lys Ile Glu His Leu Lys Thr Pro Glu Gln Asn Val Glu Leu 180 185 190 Ala Ala Ala Glu Leu Lys Ala Asp Glu Lys Ser His Ala Val Ile Leu 195 200 205 Gly Asp Thr Arg Tyr Gly Ser Glu Glu Lys Gly Thr Tyr His Leu Ala 210 215 220 Leu Phe Gly Asp Arg Ala Gln Glu Ile Ala Gly Ser Ala Thr Val Lys 225 230 235 240 Ile Gly Glu Lys Val His Glu Ile Gly Ile Ala Gly Lys Gln 245 250 <210> 87 <211> 254 <212> PRT <213> Neisseria meningitidis <400> 87 Cys Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu 1 5 10 15 Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp Lys Ser Leu Gln 20 25 30 Ser Leu Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu 35 40 45 Ala Ala Gln Gly Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn 50 55 60 Thr Gly Lys Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile Arg 65 70 75 80 Gln Ile Glu Val Asp Gly Gln Leu Ile Thr Leu Glu Ser Gly Glu Phe 85 90 95 Gln Ile Tyr Lys Gln Asp His Ser Ala Val Val Ala Leu Gln Ile Glu 100 105 110 Lys Ile Asn Asn Pro Asp Lys Ile Asp Ser Leu Ile Asn Gln Arg Ser 115 120 125 Phe Leu Val Ser Gly Leu Gly Gly Glu His Thr Ala Phe Asn Gln Leu 130 135 140 Pro Ser Gly Lys Ala Glu Tyr His Gly Lys Ala Phe Ser Ser Asp Asp 145 150 155 160 Ala Gly Gly Lys Leu Thr Tyr Thr Ile Asp Phe Ala Ala Lys Gln Gly 165 170 175 His Gly Lys Ile Glu His Leu Lys Thr Pro Glu Gln Asn Val Glu Leu 180 185 190 Ala Ser Ala Glu Leu Lys Ala Asp Glu Lys Ser His Ala Val Ile Leu 195 200 205 Gly Asp Thr Arg Tyr Gly Gly Glu Glu Lys Gly Thr Tyr His Leu Ala 210 215 220 Leu Phe Gly Asp Arg Ala Gln Glu Ile Ala Gly Ser Ala Thr Val Lys 225 230 235 240 Ile Arg Glu Lys Val His Glu Ile Gly Ile Ala Gly Lys Gln 245 250 <210> 88 <211> 254 <212> PRT <213> Neisseria meningitidis <400> 88 Cys Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu 1 5 10 15 Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp Lys Ser Leu Gln 20 25 30 Ser Leu Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu 35 40 45 Ala Ala Gln Gly Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn 50 55 60 Thr Gly Lys Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile Arg 65 70 75 80 Gln Ile Glu Val Asp Gly Gln Leu Ile Thr Leu Glu Ser Gly Glu Phe 85 90 95 Gln Ile Tyr Lys Gln Asp His Ser Ala Val Val Ala Leu Gln Ile Glu 100 105 110 Lys Ile Asn Asn Pro Asp Lys Ile Asp Ser Leu Ile Asn Gln Arg Ser 115 120 125 Phe Leu Val Ser Gly Leu Gly Gly Glu His Thr Ala Phe Asn Gln Leu 130 135 140 Pro Ser Gly Lys Ala Glu Tyr His Gly Lys Ala Phe Ser Ser Asp Asp 145 150 155 160 Pro Asn Gly Arg Leu His Tyr Ser Ile Asp Phe Thr Lys Lys Gln Gly 165 170 175 Tyr Gly Arg Ile Glu His Leu Lys Thr Pro Glu Gln Asn Val Glu Leu 180 185 190 Ala Ser Ala Glu Leu Lys Ala Asp Glu Lys Ser His Ala Val Ile Leu 195 200 205 Gly Asp Thr Arg Tyr Gly Gly Glu Glu Lys Gly Thr Tyr His Leu Ala 210 215 220 Leu Phe Gly Asp Arg Ala Gln Glu Ile Ala Gly Ser Ala Thr Val Lys 225 230 235 240 Ile Arg Glu Lys Val His Glu Ile Gly Ile Ala Gly Lys Gln 245 250 <210> 89 <211> 254 <212> PRT <213> Neisseria meningitidis <400> 89 Cys Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu 1 5 10 15 Ala Asp Ala Leu Thr Thr Pro Leu Asp His Lys Asp Lys Ser Leu Gln 20 25 30 Ser Leu Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu 35 40 45 Ala Ala Gln Gly Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn 50 55 60 Thr Gly Lys Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile Arg 65 70 75 80 Gln Ile Glu Val Asp Gly Gln Thr Ile Thr Leu Ala Ser Gly Glu Phe 85 90 95 Gln Ile Tyr Lys Gln Asn His Ser Ala Val Val Ala Leu Gln Ile Glu 100 105 110 Lys Ile Asn Asn Pro Asp Lys Ile Asp Ser Leu Ile Asn Gln Arg Ser 115 120 125 Phe Leu Val Ser Gly Leu Gly Gly Glu His Thr Ala Phe Asn Gln Leu 130 135 140 Pro Asp Gly Lys Ala Glu Tyr His Gly Lys Ala Phe Ser Ser Asp Asp 145 150 155 160 Pro Asn Gly Arg Leu His Tyr Ser Ile Asp Phe Thr Lys Lys Gln Gly 165 170 175 Tyr Gly Arg Ile Glu His Leu Lys Thr Pro Glu Gln Asn Val Glu Leu 180 185 190 Ala Ser Ala Glu Leu Lys Ala Asp Glu Lys Ser His Ala Val Ile Leu 195 200 205 Gly Asp Thr Arg Tyr Gly Gly Glu Glu Lys Gly Thr Tyr His Leu Ala 210 215 220 Leu Phe Gly Asp Arg Ala Gln Glu Ile Ala Gly Ser Ala Thr Val Lys 225 230 235 240 Ile Arg Glu Lys Val His Glu Ile Gly Ile Ala Gly Lys Gln 245 250 <210> 90 <211> 261 <212> PRT <213> Neisseria meningitidis <400> 90 Cys Ser Ser Gly Ser Gly Ser Gly Gly Gly Gly Val Ala Ala Asp Ile 1 5 10 15 Gly Thr Gly Leu Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp 20 25 30 Lys Gly Leu Lys Ser Leu Thr Leu Glu Asp Ser Ile Ser Gln Asn Gly 35 40 45 Thr Leu Thr Leu Ser Ala Gln Gly Ala Glu Lys Thr Phe Lys Val Gly 50 55 60 Asp Lys Asp Asn Ser Leu Asn Thr Gly Lys Leu Lys Asn Asp Lys Ile 65 70 75 80 Ser Arg Phe Asp Phe Val Gln Lys Ile Glu Val Asp Gly Gln Thr Ile 85 90 95 Thr Leu Ala Ser Gly Glu Phe Gln Ile Tyr Lys Gln Asp His Ser Ala 100 105 110 Val Val Ala Leu Gln Ile Glu Lys Ile Asn Asn Pro Asp Lys Ile Asp 115 120 125 Ser Leu Ile Asn Gln Arg Ser Phe Leu Val Ser Gly Leu Gly Gly Glu 130 135 140 His Thr Ala Phe Asn Gln Leu Pro Ser Gly Lys Ala Glu Tyr His Gly 145 150 155 160 Lys Ala Phe Ser Ser Asp Asp Ala Gly Gly Lys Leu Thr Tyr Thr Ile 165 170 175 Asp Phe Ala Ala Lys Gln Gly His Gly Lys Ile Glu His Leu Lys Thr 180 185 190 Pro Glu Gln Asn Val Glu Leu Ala Ser Ala Glu Leu Lys Ala Asp Glu 195 200 205 Lys Ser His Ala Val Ile Leu Gly Asp Thr Arg Tyr Gly Ser Glu Glu 210 215 220 Lys Gly Thr Tyr His Leu Ala Leu Phe Gly Asp Arg Ala Gln Glu Ile 225 230 235 240 Ala Gly Ser Ala Thr Val Lys Ile Arg Glu Lys Val His Glu Ile Gly 245 250 255 Ile Ala Gly Lys Gln 260 <210> 91 <211> 257 <212> PRT <213> Neisseria meningitidis <400> 91 Cys Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu 1 5 10 15 Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp Lys Gly Leu Lys 20 25 30 Ser Leu Thr Leu Glu Asp Ser Ile Ser Gln Asn Gly Thr Leu Thr Leu 35 40 45 Ser Ala Gln Gly Ala Glu Lys Thr Phe Lys Val Gly Asp Lys Asp Asn 50 55 60 Ser Leu Asn Thr Gly Lys Leu Lys Asn Asp Lys Ile Ser Arg Phe Asp 65 70 75 80 Phe Val Gln Lys Ile Glu Val Asp Gly Gln Thr Ile Thr Leu Ala Ser 85 90 95 Gly Glu Phe Gln Ile Tyr Lys Gln Asn His Ser Ala Val Val Ala Leu 100 105 110 Gln Ile Glu Lys Ile Asn Asn Pro Asp Lys Ile Asp Ser Leu Ile Asn 115 120 125 Gln Arg Ser Phe Leu Val Ser Gly Leu Gly Gly Glu His Thr Ala Phe 130 135 140 Asn Gln Leu Pro Gly Gly Lys Ala Glu Tyr His Gly Lys Ala Phe Ser 145 150 155 160 Ser Asp Asp Ala Gly Gly Lys Leu Thr Tyr Thr Ile Asp Phe Ala Ala 165 170 175 Lys Gln Gly His Gly Lys Ile Glu His Leu Lys Thr Pro Glu Gln Asn 180 185 190 Val Glu Leu Ala Ala Ala Glu Leu Lys Ala Asp Glu Lys Ser His Ala 195 200 205 Val Ile Leu Gly Asp Thr Arg Tyr Gly Ser Glu Glu Lys Gly Thr Tyr 210 215 220 His Leu Ala Leu Phe Gly Asp Arg Ala Gln Glu Ile Ala Gly Ser Ala 225 230 235 240 Thr Val Lys Ile Gly Glu Lys Val His Glu Ile Ser Ile Ala Gly Lys 245 250 255 Gln <210> 92 <211> 268 <212> PRT <213> Artificial Sequence <220> <223> Modified Polypeptide <400> 92 Cys Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu 1 5 10 15 Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp Lys Gly Leu Gln 20 25 30 Ser Leu Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu 35 40 45 Ala Ala Gln Gly Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn 50 55 60 Thr Gly Lys Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile Arg 65 70 75 80 Gln Ile Glu Val Asp Tyr Tyr Thr Lys Asp Thr Asn Asn Asn Leu Thr 85 90 95 Leu Val Pro Gln Leu Ile Thr Leu Glu Ser Gly Glu Phe Gln Val Tyr 100 105 110 Lys Gln Ser His Ser Ala Leu Thr Ala Phe Gln Thr Glu Gln Ile Gln 115 120 125 Asp Ser Glu His Ser Gly Lys Met Val Ala Lys Arg Gln Phe Arg Ile 130 135 140 Gly Asp Ile Ala Gly Glu His Thr Ser Phe Asp Lys Leu Pro Glu Gly 145 150 155 160 Gly Arg Ala Thr Tyr Arg Gly Thr Ala Phe Gly Ser Asp Asp Ala Gly 165 170 175 Gly Lys Leu Thr Tyr Thr Ile Asp Phe Ala Ala Lys Gln Gly Asn Gly 180 185 190 Lys Ile Glu His Leu Lys Ser Pro Glu Leu Asn Val Asp Leu Ala Ala 195 200 205 Ala Asp Ile Lys Pro Asp Gly Lys Arg His Ala Val Ile Ser Gly Ser 210 215 220 Val Leu Tyr Asn Gln Ala Glu Lys Gly Ser Tyr Ser Leu Gly Ile Phe 225 230 235 240 Gly Gly Lys Ala Gln Glu Val Ala Gly Ser Ala Glu Val Lys Thr Val 245 250 255 Asn Gly Ile Arg His Ile Gly Leu Ala Ala Lys Gln 260 265 <210> 93 <211> 274 <212> PRT <213> Artificial Sequence <220> <223> Modified Polypeptide <400> 93 Cys Ser Ser Gly Ser Gly Ser Gly Gly Gly Gly Val Ala Ala Asp Ile 1 5 10 15 Gly Thr Gly Leu Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp 20 25 30 Lys Gly Leu Lys Ser Leu Thr Leu Glu Asp Ser Ile Ser Gln Asn Gly 35 40 45 Thr Leu Thr Leu Ser Ala Gln Gly Ala Glu Lys Thr Phe Lys Val Gly 50 55 60 Asp Lys Asp Asn Ser Leu Asn Thr Gly Lys Leu Lys Asn Asp Lys Ile 65 70 75 80 Ser Arg Phe Asp Phe Val Gln Lys Ile Glu Val Asp Tyr Tyr Thr Lys 85 90 95 Asp Thr Asn Asn Asn Leu Thr Leu Val Pro Gln Thr Ile Thr Leu Ala 100 105 110 Ser Gly Glu Phe Gln Ile Tyr Lys Gln Asp His Ser Ala Val Val Ala 115 120 125 Leu Gln Ile Glu Lys Ile Asn Asn Pro Asp Lys Ile Asp Ser Leu Ile 130 135 140 Asn Gln Arg Ser Phe Leu Val Ser Gly Leu Gly Gly Glu His Thr Ala 145 150 155 160 Phe Asn Gln Leu Pro Ser Gly Lys Ala Glu Tyr His Gly Lys Ala Phe 165 170 175 Ser Ser Asp Asp Ala Gly Gly Lys Leu Thr Tyr Thr Ile Asp Phe Ala 180 185 190 Ala Lys Gln Gly His Gly Lys Ile Glu His Leu Lys Thr Pro Glu Gln 195 200 205 Asn Val Glu Leu Ala Ser Ala Glu Leu Lys Ala Asp Glu Lys Ser His 210 215 220 Ala Val Ile Leu Gly Asp Thr Arg Tyr Gly Ser Glu Glu Lys Gly Thr 225 230 235 240 Tyr His Leu Ala Leu Phe Gly Asp Arg Ala Gln Glu Ile Ala Gly Ser 245 250 255 Ala Thr Val Lys Ile Arg Glu Lys Val His Glu Ile Gly Ile Ala Gly 260 265 270 Lys Gln <210> 94 <211> 267 <212> PRT <213> Artificial Sequence <220> <223> Modified Polypeptide <400> 94 Cys Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu 1 5 10 15 Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp Lys Ser Leu Gln 20 25 30 Ser Leu Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu 35 40 45 Ala Ala Gln Gly Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn 50 55 60 Thr Gly Lys Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile Arg 65 70 75 80 Gln Ile Glu Val Asp Tyr Tyr Thr Lys Asp Thr Asn Asn Asn Leu Thr 85 90 95 Leu Val Pro Gln Leu Ile Thr Leu Glu Ser Gly Glu Phe Gln Ile Tyr 100 105 110 Lys Gln Asp His Ser Ala Val Val Ala Leu Gln Ile Glu Lys Ile Asn 115 120 125 Asn Pro Asp Lys Ile Asp Ser Leu Ile Asn Gln Arg Ser Phe Leu Val 130 135 140 Ser Gly Leu Gly Gly Glu His Thr Ala Phe Asn Gln Leu Pro Ser Gly 145 150 155 160 Lys Ala Glu Tyr His Gly Lys Ala Phe Ser Ser Asp Asp Ala Gly Gly 165 170 175 Lys Leu Thr Tyr Thr Ile Asp Phe Ala Ala Lys Gln Gly His Gly Lys 180 185 190 Ile Glu His Leu Lys Thr Pro Glu Gln Asn Val Glu Leu Ala Ser Ala 195 200 205 Glu Leu Lys Ala Asp Glu Lys Ser His Ala Val Ile Leu Gly Asp Thr 210 215 220 Arg Tyr Gly Gly Glu Glu Lys Gly Thr Tyr His Leu Ala Leu Phe Gly 225 230 235 240 Asp Arg Ala Gln Glu Ile Ala Gly Ser Ala Thr Val Lys Ile Arg Glu 245 250 255 Lys Val His Glu Ile Gly Ile Ala Gly Lys Gln 260 265 <210> 95 <211> 268 <212> PRT <213> Artificial Sequence <220> <223> Modified Polypeptide <400> 95 Cys Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu 1 5 10 15 Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp Lys Gly Leu Gln 20 25 30 Ser Leu Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu 35 40 45 Ala Ala Gln Gly Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn 50 55 60 Thr Gly Lys Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile Arg 65 70 75 80 Gln Ile Glu Val Asp Gly Gln Leu Ile Thr Leu Glu Ser Gly Glu Phe 85 90 95 Gln Val Tyr Lys Gln Ser His Ser Ala Leu Thr Ala Phe Gln Thr Glu 100 105 110 Gln Ile Gln Asp Ser Glu His Ser Gly Lys Met Val Ala Lys Arg Gln 115 120 125 Phe Arg Ile Gly Asp Ile Ala Gly Glu His Thr Ser Phe Asp Lys Leu 130 135 140 Pro Glu Gly Gly Arg Ala Thr Tyr Arg Gly Thr Ala Phe Gly Ser Asp 145 150 155 160 Asp Ala Gly Gly Lys Leu Thr Tyr Thr Ile Asp Phe Ala Ala Lys Gln 165 170 175 Gly Asn Gly Lys Ile Glu His Leu Lys Ser Pro Glu Leu Asn Val Asp 180 185 190 Leu Ala Ala Ala Asp Ile Lys Pro Asp Tyr Tyr Thr Lys Asp Thr Asn 195 200 205 Asn Asn Leu Thr Leu Val Pro Lys Arg His Ala Val Ile Ser Gly Ser 210 215 220 Val Leu Tyr Asn Gln Ala Glu Lys Gly Ser Tyr Ser Leu Gly Ile Phe 225 230 235 240 Gly Gly Lys Ala Gln Glu Val Ala Gly Ser Ala Glu Val Lys Thr Val 245 250 255 Asn Gly Ile Arg His Ile Gly Leu Ala Ala Lys Gln 260 265 <210> 96 <211> 274 <212> PRT <213> Artificial Sequence <220> <223> Modified Polypeptide <400> 96 Cys Ser Ser Gly Ser Gly Ser Gly Gly Gly Gly Val Ala Ala Asp Ile 1 5 10 15 Gly Thr Gly Leu Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp 20 25 30 Lys Gly Leu Lys Ser Leu Thr Leu Glu Asp Ser Ile Ser Gln Asn Gly 35 40 45 Thr Leu Thr Leu Ser Ala Gln Gly Ala Glu Lys Thr Phe Lys Val Gly 50 55 60 Asp Lys Asp Asn Ser Leu Asn Thr Gly Lys Leu Lys Asn Asp Lys Ile 65 70 75 80 Ser Arg Phe Asp Phe Val Gln Lys Ile Glu Val Asp Gly Gln Thr Ile 85 90 95 Thr Leu Ala Ser Gly Glu Phe Gln Ile Tyr Lys Gln Asp His Ser Ala 100 105 110 Val Val Ala Leu Gln Ile Glu Lys Ile Asn Asn Pro Asp Lys Ile Asp 115 120 125 Ser Leu Ile Asn Gln Arg Ser Phe Leu Val Ser Gly Leu Gly Gly Glu 130 135 140 His Thr Ala Phe Asn Gln Leu Pro Ser Gly Lys Ala Glu Tyr His Gly 145 150 155 160 Lys Ala Phe Ser Ser Asp Asp Ala Gly Gly Lys Leu Thr Tyr Thr Ile 165 170 175 Asp Phe Ala Ala Lys Gln Gly His Gly Lys Ile Glu His Leu Lys Thr 180 185 190 Pro Glu Gln Asn Val Glu Leu Ala Ser Ala Glu Leu Lys Ala Asp Tyr 195 200 205 Tyr Thr Lys Asp Thr Asn Asn Asn Leu Thr Leu Val Pro Lys Ser His 210 215 220 Ala Val Ile Leu Gly Asp Thr Arg Tyr Gly Ser Glu Glu Lys Gly Thr 225 230 235 240 Tyr His Leu Ala Leu Phe Gly Asp Arg Ala Gln Glu Ile Ala Gly Ser 245 250 255 Ala Thr Val Lys Ile Arg Glu Lys Val His Glu Ile Gly Ile Ala Gly 260 265 270 Lys Gln <210> 97 <211> 267 <212> PRT <213> Artificial sequence <220> <223> Modified polypeptide <400> 97 Cys Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu 1 5 10 15 Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp Lys Ser Leu Gln 20 25 30 Ser Leu Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu 35 40 45 Ala Ala Gln Gly Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn 50 55 60 Thr Gly Lys Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile Arg 65 70 75 80 Gln Ile Glu Val Asp Gly Gln Leu Ile Thr Leu Glu Ser Gly Glu Phe 85 90 95 Gln Ile Tyr Lys Gln Asp His Ser Ala Val Val Ala Leu Gln Ile Glu 100 105 110 Lys Ile Asn Asn Pro Asp Lys Ile Asp Ser Leu Ile Asn Gln Arg Ser 115 120 125 Phe Leu Val Ser Gly Leu Gly Gly Glu His Thr Ala Phe Asn Gln Leu 130 135 140 Pro Ser Gly Lys Ala Glu Tyr His Gly Lys Ala Phe Ser Ser Asp Asp 145 150 155 160 Ala Gly Gly Lys Leu Thr Tyr Thr Ile Asp Phe Ala Ala Lys Gln Gly 165 170 175 His Gly Lys Ile Glu His Leu Lys Thr Pro Glu Gln Asn Val Glu Leu 180 185 190 Ala Ser Ala Glu Leu Lys Ala Asp Tyr Tyr Thr Lys Asp Thr Asn Asn 195 200 205 Asn Leu Thr Leu Val Pro Lys Ser His Ala Val Ile Leu Gly Asp Thr 210 215 220 Arg Tyr Gly Gly Glu Glu Lys Gly Thr Tyr His Leu Ala Leu Phe Gly 225 230 235 240 Asp Arg Ala Gln Glu Ile Ala Gly Ser Ala Thr Val Lys Ile Arg Glu 245 250 255 Lys Val His Glu Ile Gly Ile Ala Gly Lys Gln 260 265 <210> 98 <211> 268 <212> PRT <213> Artificial sequence <220> <223> Modified polypeptide <400> 98 Cys Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu 1 5 10 15 Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp Lys Gly Leu Gln 20 25 30 Ser Leu Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu 35 40 45 Ala Ala Gln Gly Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn 50 55 60 Thr Gly Lys Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile Arg 65 70 75 80 Gln Ile Glu Val Asp Gly Gln Leu Ile Thr Leu Glu Ser Gly Glu Phe 85 90 95 Gln Val Tyr Lys Gln Ser His Ser Ala Leu Thr Ala Phe Gln Thr Glu 100 105 110 Gln Ile Gln Asp Ser Glu His Ser Gly Lys Met Val Ala Lys Arg Gln 115 120 125 Phe Arg Ile Gly Asp Ile Ala Gly Glu His Thr Ser Phe Asp Lys Leu 130 135 140 Pro Glu Gly Gly Arg Ala Thr Tyr Arg Gly Thr Ala Phe Gly Ser Asp 145 150 155 160 Asp Ala Gly Gly Lys Leu Thr Tyr Thr Ile Asp Phe Ala Ala Lys Gln 165 170 175 Gly Asn Gly Lys Ile Glu His Leu Lys Ser Pro Glu Leu Asn Val Asp 180 185 190 Leu Ala Ala Ala Asp Ile Lys Pro Asp Gly Lys Arg His Ala Val Ile 195 200 205 Ser Gly Ser Val Leu Tyr Asn Gln Ala Glu Lys Gly Ser Tyr Ser Leu 210 215 220 Gly Ile Phe Gly Tyr Tyr Thr Lys Asp Thr Asn Asn Asn Leu Thr Leu 225 230 235 240 Val Pro Lys Ala Gln Glu Val Ala Gly Ser Ala Glu Val Lys Thr Val 245 250 255 Asn Gly Ile Arg His Ile Gly Leu Ala Ala Lys Gln 260 265 <210> 99 <211> 274 <212> PRT <213> Artificial Sequence <220> <223> Modified Polypeptide <400> 99 Cys Ser Ser Gly Ser Gly Ser Gly Gly Gly Gly Val Ala Ala Asp Ile 1 5 10 15 Gly Thr Gly Leu Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp 20 25 30 Lys Gly Leu Lys Ser Leu Thr Leu Glu Asp Ser Ile Ser Gln Asn Gly 35 40 45 Thr Leu Thr Leu Ser Ala Gln Gly Ala Glu Lys Thr Phe Lys Val Gly 50 55 60 Asp Lys Asp Asn Ser Leu Asn Thr Gly Lys Leu Lys Asn Asp Lys Ile 65 70 75 80 Ser Arg Phe Asp Phe Val Gln Lys Ile Glu Val Asp Gly Gln Thr Ile 85 90 95 Thr Leu Ala Ser Gly Glu Phe Gln Ile Tyr Lys Gln Asp His Ser Ala 100 105 110 Val Val Ala Leu Gln Ile Glu Lys Ile Asn Asn Pro Asp Lys Ile Asp 115 120 125 Ser Leu Ile Asn Gln Arg Ser Phe Leu Val Ser Gly Leu Gly Gly Glu 130 135 140 His Thr Ala Phe Asn Gln Leu Pro Ser Gly Lys Ala Glu Tyr His Gly 145 150 155 160 Lys Ala Phe Ser Ser Asp Asp Ala Gly Gly Lys Leu Thr Tyr Thr Ile 165 170 175 Asp Phe Ala Ala Lys Gln Gly His Gly Lys Ile Glu His Leu Lys Thr 180 185 190 Pro Glu Gln Asn Val Glu Leu Ala Ser Ala Glu Leu Lys Ala Asp Glu 195 200 205 Lys Ser His Ala Val Ile Leu Gly Asp Thr Arg Tyr Gly Ser Glu Glu 210 215 220 Lys Gly Thr Tyr His Leu Ala Leu Phe Gly Tyr Tyr Thr Lys Asp Thr 225 230 235 240 Asn Asn Asn Leu Thr Leu Val Pro Arg Ala Gln Glu Ile Ala Gly Ser 245 250 255 Ala Thr Val Lys Ile Arg Glu Lys Val His Glu Ile Gly Ile Ala Gly 260 265 270 Lys Gln <210> 100 <211> 267 <212> PRT <213> Artificial Sequence <220> <223> Modified Polypeptide <400> 100 Cys Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu 1 5 10 15 Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp Lys Ser Leu Gln 20 25 30 Ser Leu Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu 35 40 45 Ala Ala Gln Gly Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn 50 55 60 Thr Gly Lys Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile Arg 65 70 75 80 Gln Ile Glu Val Asp Gly Gln Leu Ile Thr Leu Glu Ser Gly Glu Phe 85 90 95 Gln Ile Tyr Lys Gln Asp His Ser Ala Val Val Ala Leu Gln Ile Glu 100 105 110 Lys Ile Asn Asn Pro Asp Lys Ile Asp Ser Leu Ile Asn Gln Arg Ser 115 120 125 Phe Leu Val Ser Gly Leu Gly Gly Glu His Thr Ala Phe Asn Gln Leu 130 135 140 Pro Ser Gly Lys Ala Glu Tyr His Gly Lys Ala Phe Ser Ser Asp Asp 145 150 155 160 Ala Gly Gly Lys Leu Thr Tyr Thr Ile Asp Phe Ala Ala Lys Gln Gly 165 170 175 His Gly Lys Ile Glu His Leu Lys Thr Pro Glu Gln Asn Val Glu Leu 180 185 190 Ala Ser Ala Glu Leu Lys Ala Asp Glu Lys Ser His Ala Val Ile Leu 195 200 205 Gly Asp Thr Arg Tyr Gly Gly Glu Glu Lys Gly Thr Tyr His Leu Ala 210 215 220 Leu Phe Gly Tyr Tyr Thr Lys Asp Thr Asn Asn Asn Leu Thr Leu Val 225 230 235 240 Pro Arg Ala Gln Glu Ile Ala Gly Ser Ala Thr Val Lys Ile Arg Glu 245 250 255 Lys Val His Glu Ile Gly Ile Ala Gly Lys Gln 260 265 <210> 101 <211> 268 <212> PRT <213> Synthetic Sequence <220> <223> Modified Polypeptide <400> 101 Cys Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu 1 5 10 15 Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp Lys Gly Leu Gln 20 25 30 Ser Leu Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu 35 40 45 Ala Ala Gln Tyr Tyr Thr Lys Asp Thr Asn Asn Asn Leu Thr Leu Val 50 55 60 Pro Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn Thr Gly Lys 65 70 75 80 Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile Arg Gln Ile Glu 85 90 95 Val Asp Gly Gln Leu Ile Thr Leu Glu Ser Gly Glu Phe Gln Val Tyr 100 105 110 Lys Gln Ser His Ser Ala Leu Thr Ala Phe Gln Thr Glu Gln Ile Gln 115 120 125 Asp Ser Glu His Ser Gly Lys Met Val Ala Lys Arg Gln Phe Arg Ile 130 135 140 Gly Asp Ile Ala Gly Glu His Thr Ser Phe Asp Lys Leu Pro Glu Gly 145 150 155 160 Gly Arg Ala Thr Tyr Arg Gly Thr Ala Phe Gly Ser Asp Asp Ala Gly 165 170 175 Gly Lys Leu Thr Tyr Thr Ile Asp Phe Ala Ala Lys Gln Gly Asn Gly 180 185 190 Lys Ile Glu His Leu Lys Ser Pro Glu Leu Asn Val Asp Leu Ala Ala 195 200 205 Ala Asp Ile Lys Pro Asp Gly Lys Arg His Ala Val Ile Ser Gly Ser 210 215 220 Val Leu Tyr Asn Gln Ala Glu Lys Gly Ser Tyr Ser Leu Gly Ile Phe 225 230 235 240 Gly Gly Lys Ala Gln Glu Val Ala Gly Ser Ala Glu Val Lys Thr Val 245 250 255 Asn Gly Ile Arg His Ile Gly Leu Ala Ala Lys Gln 260 265 <210> 102 <211> 274 <212> PRT <213> Artificial sequence <220> <223> Modified polypeptide <400> 102 Cys Ser Ser Gly Ser Gly Ser Gly Gly Gly Gly Val Ala Ala Asp Ile 1 5 10 15 Gly Thr Gly Leu Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp 20 25 30 Lys Gly Leu Lys Ser Leu Thr Leu Glu Asp Ser Ile Ser Gln Asn Gly 35 40 45 Thr Leu Thr Leu Ser Ala Gln Tyr Tyr Thr Lys Asp Thr Asn Asn Asn 50 55 60 Leu Thr Leu Val Pro Ala Glu Lys Thr Phe Lys Val Gly Asp Lys Asp 65 70 75 80 Asn Ser Leu Asn Thr Gly Lys Leu Lys Asn Asp Lys Ile Ser Arg Phe 85 90 95 Asp Phe Val Gln Lys Ile Glu Val Asp Gly Gln Thr Ile Thr Leu Ala 100 105 110 Ser Gly Glu Phe Gln Ile Tyr Lys Gln Asp His Ser Ala Val Val Ala 115 120 125 Leu Gln Ile Glu Lys Ile Asn Asn Pro Asp Lys Ile Asp Ser Leu Ile 130 135 140 Asn Gln Arg Ser Phe Leu Val Ser Gly Leu Gly Gly Glu His Thr Ala 145 150 155 160 Phe Asn Gln Leu Pro Ser Gly Lys Ala Glu Tyr His Gly Lys Ala Phe 165 170 175 Ser Ser Asp Asp Ala Gly Gly Lys Leu Thr Tyr Thr Ile Asp Phe Ala 180 185 190 Ala Lys Gln Gly His Gly Lys Ile Glu His Leu Lys Thr Pro Glu Gln 195 200 205 Asn Val Glu Leu Ala Ser Ala Glu Leu Lys Ala Asp Glu Lys Ser His 210 215 220 Ala Val Ile Leu Gly Asp Thr Arg Tyr Gly Ser Glu Glu Lys Gly Thr 225 230 235 240 Tyr His Leu Ala Leu Phe Gly Asp Arg Ala Gln Glu Ile Ala Gly Ser 245 250 255 Ala Thr Val Lys Ile Arg Glu Lys Val His Glu Ile Gly Ile Ala Gly 260 265 270 Lys Gln <210> 103 <211> 267 <212> PRT <213> Artificial Sequence <220> <223> Modified Polypeptide <400> 103 Cys Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu 1 5 10 15 Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp Lys Ser Leu Gln 20 25 30 Ser Leu Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu 35 40 45 Ala Ala Gln Tyr Tyr Thr Lys Asp Thr Asn Asn Asn Leu Thr Leu Val 50 55 60 Pro Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn Thr Gly Lys 65 70 75 80 Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile Arg Gln Ile Glu 85 90 95 Val Asp Gly Gln Leu Ile Thr Leu Glu Ser Gly Glu Phe Gln Ile Tyr 100 105 110 Lys Gln Asp His Ser Ala Val Val Ala Leu Gln Ile Glu Lys Ile Asn 115 120 125 Asn Pro Asp Lys Ile Asp Ser Leu Ile Asn Gln Arg Ser Phe Leu Val 130 135 140 Ser Gly Leu Gly Gly Glu His Thr Ala Phe Asn Gln Leu Pro Ser Gly 145 150 155 160 Lys Ala Glu Tyr His Gly Lys Ala Phe Ser Ser Asp Asp Ala Gly Gly 165 170 175 Lys Leu Thr Tyr Thr Ile Asp Phe Ala Ala Lys Gln Gly His Gly Lys 180 185 190 Ile Glu His Leu Lys Thr Pro Glu Gln Asn Val Glu Leu Ala Ser Ala 195 200 205 Glu Leu Lys Ala Asp Glu Lys Ser His Ala Val Ile Leu Gly Asp Thr 210 215 220 Arg Tyr Gly Gly Glu Glu Lys Gly Thr Tyr His Leu Ala Leu Phe Gly 225 230 235 240 Asp Arg Ala Gln Glu Ile Ala Gly Ser Ala Thr Val Lys Ile Arg Glu 245 250 255 Lys Val His Glu Ile Gly Ile Ala Gly Lys Gln 260 265 <210> 104 <211> 268 <212> PRT <213> Artificial Sequence <220> <223> Modified Polypeptide <400> 104 Cys Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu 1 5 10 15 Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp Lys Gly Leu Gln 20 25 30 Ser Leu Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu 35 40 45 Ala Ala Gln Gly Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn 50 55 60 Thr Gly Lys Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile Arg 65 70 75 80 Gln Ile Glu Val Asp Gly Gln Leu Ile Thr Leu Glu Ser Gly Glu Phe 85 90 95 Gln Val Tyr Lys Gln Ser His Ser Ala Leu Thr Ala Phe Gln Thr Glu 100 105 110 Gln Ile Gln Asp Ser Tyr Tyr Thr Lys Asp Thr Asn Asn Asn Leu Thr 115 120 125 Leu Val Pro His Ser Gly Lys Met Val Ala Lys Arg Gln Phe Arg Ile 130 135 140 Gly Asp Ile Ala Gly Glu His Thr Ser Phe Asp Lys Leu Pro Glu Gly 145 150 155 160 Gly Arg Ala Thr Tyr Arg Gly Thr Ala Phe Gly Ser Asp Asp Ala Gly 165 170 175 Gly Lys Leu Thr Tyr Thr Ile Asp Phe Ala Ala Lys Gln Gly Asn Gly 180 185 190 Lys Ile Glu His Leu Lys Ser Pro Glu Leu Asn Val Asp Leu Ala Ala 195 200 205 Ala Asp Ile Lys Pro Asp Gly Lys Arg His Ala Val Ile Ser Gly Ser 210 215 220 Val Leu Tyr Asn Gln Ala Glu Lys Gly Ser Tyr Ser Leu Gly Ile Phe 225 230 235 240 Gly Gly Lys Ala Gln Glu Val Ala Gly Ser Ala Glu Val Lys Thr Val 245 250 255 Asn Gly Ile Arg His Ile Gly Leu Ala Ala Lys Gln 260 265 <210> 105 <211> 274 <212> PRT <213> Artificial sequence <220> <223> Modified polypeptide <400> 105 Cys Ser Ser Gly Ser Gly Ser Gly Gly Gly Gly Val Ala Ala Asp Ile 1 5 10 15 Gly Thr Gly Leu Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp 20 25 30 Lys Gly Leu Lys Ser Leu Thr Leu Glu Asp Ser Ile Ser Gln Asn Gly 35 40 45 Thr Leu Thr Leu Ser Ala Gln Gly Ala Glu Lys Thr Phe Lys Val Gly 50 55 60 Asp Lys Asp Asn Ser Leu Asn Thr Gly Lys Leu Lys Asn Asp Lys Ile 65 70 75 80 Ser Arg Phe Asp Phe Val Gln Lys Ile Glu Val Asp Gly Gln Thr Ile 85 90 95 Thr Leu Ala Ser Gly Glu Phe Gln Ile Tyr Lys Gln Asp His Ser Ala 100 105 110 Val Val Ala Leu Gln Ile Glu Lys Ile Asn Asn Pro Tyr Tyr Thr Lys 115 120 125 Asp Thr Asn Asn Asn Leu Thr Leu Val Pro Lys Ile Asp Ser Leu Ile 130 135 140 Asn Gln Arg Ser Phe Leu Val Ser Gly Leu Gly Gly Glu His Thr Ala 145 150 155 160 Phe Asn Gln Leu Pro Ser Gly Lys Ala Glu Tyr His Gly Lys Ala Phe 165 170 175 Ser Ser Asp Asp Ala Gly Gly Lys Leu Thr Tyr Thr Ile Asp Phe Ala 180 185 190 Ala Lys Gln Gly His Gly Lys Ile Glu His Leu Lys Thr Pro Glu Gln 195 200 205 Asn Val Glu Leu Ala Ser Ala Glu Leu Lys Ala Asp Glu Lys Ser His 210 215 220 Ala Val Ile Leu Gly Asp Thr Arg Tyr Gly Ser Glu Glu Lys Gly Thr 225 230 235 240 Tyr His Leu Ala Leu Phe Gly Asp Arg Ala Gln Glu Ile Ala Gly Ser 245 250 255 Ala Thr Val Lys Ile Arg Glu Lys Val His Glu Ile Gly Ile Ala Gly 260 265 270 Lys Gln <210> 106 <211> 267 <212> PRT <213> Artificial Sequence <220> <223> Modified Polypeptide <400> 106 Cys Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu 1 5 10 15 Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp Lys Ser Leu Gln 20 25 30 Ser Leu Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu 35 40 45 Ala Ala Gln Gly Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn 50 55 60 Thr Gly Lys Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile Arg 65 70 75 80 Gln Ile Glu Val Asp Gly Gln Leu Ile Thr Leu Glu Ser Gly Glu Phe 85 90 95 Gln Ile Tyr Lys Gln Asp His Ser Ala Val Val Ala Leu Gln Ile Glu 100 105 110 Lys Ile Asn Asn Pro Tyr Tyr Thr Lys Asp Thr Asn Asn Asn Leu Thr 115 120 125 Leu Val Pro Lys Ile Asp Ser Leu Ile Asn Gln Arg Ser Phe Leu Val 130 135 140 Ser Gly Leu Gly Gly Glu His Thr Ala Phe Asn Gln Leu Pro Ser Gly 145 150 155 160 Lys Ala Glu Tyr His Gly Lys Ala Phe Ser Ser Asp Asp Ala Gly Gly 165 170 175 Lys Leu Thr Tyr Thr Ile Asp Phe Ala Ala Lys Gln Gly His Gly Lys 180 185 190 Ile Glu His Leu Lys Thr Pro Glu Gln Asn Val Glu Leu Ala Ser Ala 195 200 205 Glu Leu Lys Ala Asp Glu Lys Ser His Ala Val Ile Leu Gly Asp Thr 210 215 220 Arg Tyr Gly Gly Glu Glu Lys Gly Thr Tyr His Leu Ala Leu Phe Gly 225 230 235 240 Asp Arg Ala Gln Glu Ile Ala Gly Ser Ala Thr Val Lys Ile Arg Glu 245 250 255 Lys Val His Glu Ile Gly Ile Ala Gly Lys Gln 260 265 <210> 107 <211> 268 <212> PRT <213> Artificial Sequence <220> <223> Modified Polypeptide <400> 107 Cys Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu 1 5 10 15 Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp Lys Gly Leu Gln 20 25 30 Ser Leu Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu 35 40 45 Ala Ala Gln Gly Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn 50 55 60 Thr Gly Lys Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile Arg 65 70 75 80 Gln Ile Glu Val Asp Gly Gln Leu Ile Thr Leu Glu Ser Gly Glu Phe 85 90 95 Gln Val Tyr Lys Gln Ser His Ser Ala Leu Thr Ala Phe Gln Thr Glu 100 105 110 Gln Ile Gln Asp Ser Glu His Ser Gly Lys Met Val Ala Lys Arg Gln 115 120 125 Phe Arg Ile Gly Asp Ile Ala Gly Glu His Thr Ser Phe Asp Lys Leu 130 135 140 Pro Glu Gly Gly Arg Ala Thr Tyr Arg Gly Thr Ala Phe Gly Ser Asp 145 150 155 160 Asp Ala Gly Gly Lys Leu Thr Tyr Thr Ile Asp Phe Ala Ala Lys Gln 165 170 175 Gly Asn Gly Lys Ile Glu His Leu Lys Ser Pro Glu Leu Asn Val Asp 180 185 190 Leu Ala Ala Ala Asp Ile Lys Pro Asp Gly Lys Arg His Ala Val Ile 195 200 205 Ser Gly Ser Val Leu Tyr Asn Gln Ala Glu Lys Gly Ser Tyr Ser Leu 210 215 220 Gly Ile Phe Gly Gly Lys Ala Gln Glu Val Ala Gly Ser Ala Glu Val 225 230 235 240 Lys Thr Val Tyr Tyr Thr Lys Asp Thr Asn Asn Asn Leu Thr Leu Val 245 250 255 Pro Gly Ile Arg His Ile Gly Leu Ala Ala Lys Gln 260 265 <210> 108 <211> 274 <212> PRT <213> Artificial Sequence <220> <223> Modified Polypeptide <400> 108 Cys Ser Ser Gly Ser Gly Ser Gly Gly Gly Gly Val Ala Ala Asp Ile 1 5 10 15 Gly Thr Gly Leu Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp 20 25 30 Lys Gly Leu Lys Ser Leu Thr Leu Glu Asp Ser Ile Ser Gln Asn Gly 35 40 45 Thr Leu Thr Leu Ser Ala Gln Gly Ala Glu Lys Thr Phe Lys Val Gly 50 55 60 Asp Lys Asp Asn Ser Leu Asn Thr Gly Lys Leu Lys Asn Asp Lys Ile 65 70 75 80 Ser Arg Phe Asp Phe Val Gln Lys Ile Glu Val Asp Gly Gln Thr Ile 85 90 95 Thr Leu Ala Ser Gly Glu Phe Gln Ile Tyr Lys Gln Asp His Ser Ala 100 105 110 Val Val Ala Leu Gln Ile Glu Lys Ile Asn Asn Pro Asp Lys Ile Asp 115 120 125 Ser Leu Ile Asn Gln Arg Ser Phe Leu Val Ser Gly Leu Gly Gly Glu 130 135 140 His Thr Ala Phe Asn Gln Leu Pro Ser Gly Lys Ala Glu Tyr His Gly 145 150 155 160 Lys Ala Phe Ser Ser Asp Asp Ala Gly Gly Lys Leu Thr Tyr Thr Ile 165 170 175 Asp Phe Ala Ala Lys Gln Gly His Gly Lys Ile Glu His Leu Lys Thr 180 185 190 Pro Glu Gln Asn Val Glu Leu Ala Ser Ala Glu Leu Lys Ala Asp Glu 195 200 205 Lys Ser His Ala Val Ile Leu Gly Asp Thr Arg Tyr Gly Ser Glu Glu 210 215 220 Lys Gly Thr Tyr His Leu Ala Leu Phe Gly Asp Arg Ala Gln Glu Ile 225 230 235 240 Ala Gly Ser Ala Thr Val Lys Ile Arg Tyr Tyr Thr Lys Asp Thr Asn 245 250 255 Asn Asn Leu Thr Leu Val Pro Lys Val His Glu Ile Gly Ile Ala Gly 260 265 270 Lys Gln <210> 109 <211> 267 <212> PRT <213> Artificial Sequence <220> <223> Modified Polypeptide <400> 109 Cys Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu 1 5 10 15 Ala Asp Ala Leu Thr Ala Pro Leu Asp His Lys Asp Lys Ser Leu Gln 20 25 30 Ser Leu Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu 35 40 45 Ala Ala Gln Gly Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn 50 55 60 Thr Gly Lys Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile Arg 65 70 75 80 Gln Ile Glu Val Asp Gly Gln Leu Ile Thr Leu Glu Ser Gly Glu Phe 85 90 95 Gln Ile Tyr Lys Gln Asp His Ser Ala Val Val Ala Leu Gln Ile Glu 100 105 110 Lys Ile Asn Asn Pro Asp Lys Ile Asp Ser Leu Ile Asn Gln Arg Ser 115 120 125 Phe Leu Val Ser Gly Leu Gly Gly Glu His Thr Ala Phe Asn Gln Leu 130 135 140 Pro Ser Gly Lys Ala Glu Tyr His Gly Lys Ala Phe Ser Ser Asp Asp 145 150 155 160 Ala Gly Gly Lys Leu Thr Tyr Thr Ile Asp Phe Ala Ala Lys Gln Gly 165 170 175 His Gly Lys Ile Glu His Leu Lys Thr Pro Glu Gln Asn Val Glu Leu 180 185 190 Ala Ser Ala Glu Leu Lys Ala Asp Glu Lys Ser His Ala Val Ile Leu 195 200 205 Gly Asp Thr Arg Tyr Gly Gly Glu Glu Lys Gly Thr Tyr His Leu Ala 210 215 220 Leu Phe Gly Asp Arg Ala Gln Glu Ile Ala Gly Ser Ala Thr Val Lys 225 230 235 240 Ile Arg Tyr Tyr Thr Lys Asp Thr Asn Asn Asn Leu Thr Leu Val Pro 245 250 255 Lys Val His Glu Ile Gly Ile Ala Gly Lys Gln 260 265

Claims

1. A modified factor H binding protein (fHbp), comprising a wild-type fHbp having an amino acid sequence of any one of SEQ ID NOs: 1-2 and 10-11, further modified by adding any one of the PorA loops, wherein the amino acid sequence of the PorA loop is as shown in any one of SEQ ID NOs: 50, 51, 53, 62, 65 and 68, wherein the PorA loop is inserted at an amino acid position corresponding to any of amino acid residue numbers 49-54, 83-88, 114-124, 199-206, 227-233 and 240-246 of SEQ ID NO: 1, or the equivalent residue numbers of any of SEQ ID NOs: 2 and 10-11.

2. The modified factor H binding protein according to claim 1, wherein The modified fHbp is modified such that it is unable to bind factor H or at least has reduced factor H binding activity.

3. A nucleic acid encoding the modified fHbp according to claim 1 or 2.

4. The nucleic acid according to claim 3, wherein The nucleic acid is a vector.

5. The nucleic acid according to claim 4, wherein The vector is a viral vector.

6. A composition comprising the modified fHbp according to claim 1 or 2 or the nucleic acid according to any one of claims 3-5.

7. The composition according to claim 6, wherein The composition comprises two or more different modified fHbp molecules.

8. The composition according to claim 6, wherein The composition comprises a pharmaceutically acceptable carrier.

9. The composition according to claim 6, wherein The composition further comprises an adjuvant.

10. The composition according to claim 6, wherein The composition may also comprise at least one other prophylactically or therapeutically active molecule.

11. The composition according to claim 10, wherein The at least one other prophylactically or therapeutically active molecule comprises: Protein: encapsulated polysaccharide vaccine; or A conjugate vaccine, wherein an antigen comprising an fHbp scaffold carrying an exogenous peptide loop is incorporated into the conjugate vaccine as a protein carrier molecule.

12. The modified fHbp according to claim 1 or 2, the nucleic acid according to any one of claims 3-5, or the composition according to any one of claims 6-11 for use as a medicament.

13. The modified fHbp according to claim 1 or 2, the nucleic acid according to any one of claims 3-5, or the composition according to any one of claims 6-11, for use in treating or preventing pathogenic infection or colonization of a subject.

14. The modified fHbp of claim 1 or 2, the nucleic acid of any one of claims 3-5, or the composition of any one of claims 6-11 for use in vaccinating a subject.

15. A combination of the modified fHbp according to claim 1 or 2, the nucleic acid according to any one of claims 3-5, or the composition according to any one of claims 6-11 and at least one other prophylactically or therapeutically active molecule.

16. The combination according to claim 15, wherein: The at least one other prophylactically or therapeutically active molecule comprises a protein:capsule polysaccharide vaccine.

17. The combination according to claim 16 or the composition according to claim 11, wherein The protein:capsule polysaccharide vaccine comprises any serogroup C or A capsule with bacterial toxoids, a bivalent vaccine of serogroup C and A capsular polysaccharides conjugated to bacterial toxoids, a tetravalent conjugate vaccine of serogroups A, C, Y, W polysaccharides conjugated to bacterial toxoids, or a pentavalent conjugate vaccine of serogroups A, C, Y, W, X polysaccharides conjugated to bacterial toxoids.

18. The combination according to claim 15 or the composition according to claim 10, wherein The at least one other prophylactically or therapeutically active molecule comprises a conjugate vaccine, wherein an antigen comprising an fHbp scaffold carrying an exogenous peptide loop is incorporated into the conjugate vaccine as a protein carrier molecule.

19. The combination or composition according to claim 18, wherein: The conjugate vaccine comprises any serogroup capsular polysaccharide selected from A, C, Y, W or X strains or a combination thereof.

20. Use of the factor H binding protein (fHbp) according to claim 1 or 2 in preparing an epitope display scaffold.

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