The invention relates to a modified V2 fHbp having increased stability over a wild type V2 fHbp; a modified V2 fHbp having an amino acid sequence with at least 85% identity to the sequence of Seq ID No: 1, wherein both Ser 137 and Gly 138 are mutated or both Val112 and Leu114 are mutated; immunogenic, pharmaceutical and vaccine compositions; nucleic acid and a host cell; and methods of use of such compositions.
1. A composition comprising:
a modified Variant 2 factor H binding protein (V2 fHbp) having increased stability over a wild type V2 fHbp, wherein the modified variant comprises one or more amino acid substitutions in the N terminal beta barrel of SEQ ID NO: 1 or a V2 fHbp protein with at least 90% identity to SEQ ID NO: 1, wherein the modified variant comprises one or more amino acid substitutions at position 30, 31, 37, 38, 41, 102, 107, 109, 132, and 133 of SEQ ID NO: 1.
2. The composition of claim 1 wherein the maximum value of the left hand peak in a differential scanning calorimetry (DSC) analysis profile of a V2 fHbp is increased by at least 5° C. compared with a wild-type fHbp.
3. The composition of claim 1 wherein the modified variant comprises a mutation at Ser 132 and Gly 133 .
4. The composition of claim 1 wherein the modified variant comprises Ser 132 Gly and Gly 133 Asp.
5. The composition of claim 1 wherein the modified variant comprises a mutation at Ser30, Asp102, Ser132 and Gly133; and wherein the mutations comprise Ser 30 Gly, Asp 102 Ser, Ser 132 Gly and Gly 133 Asp.
6. The composition of claim 1 wherein the modified variant comprises a mutation at Ser30, Leu109, Ser132 and Gly133; and wherein the mutations comprise Ser 30 Gly, Leu 109 Phe, Ser 132 Gly and Gly 133 Asp.
7. The composition of claim 1 , wherein the modified variant comprises a mutation at Ser30, Asp102, Leu109, Ser132 and Gly133; and wherein the mutations comprise Ser 35 Gly, Asp 107 Ser, Leu 114 Phe, Ser 137 Gly and Gly 138 Asp.
8. The composition of claim 1 , wherein the modified variant comprises a mutation at Ser30, Asp102, Val107, Leu109, Ser132 and Gly133; and wherein the mutations comprise Ser 30 Gly, Asp 102 Ser, Val 107 Thr, Leu 109 Phe, Ser 132 Gly and Gly 133 Asp.
9. The composition of claim 1 , wherein the modified variant comprises a mutation at Val107 and Leu109; and wherein the mutations comprise Val 107 Thr, and Leu 109 Phe.
10. An immunogenic composition comprising:
a modified V2 fHbp according to claim 1 .
11. The immunogenic composition of claim 10 , wherein the composition is capable of eliciting a bactericidal antibody response to N. meningitidis.
12. The immunogenic composition of claim 10 further comprising one or more antigens in addition to the modified V2 fHbp.
13. A therapeutic vaccine directed to N. meningitidis comprising the modified V2 fHbp of claim 1 .
14. A pharmaceutical composition comprising:
the modified V2 fHbp of claim 1 and a pharmaceutically acceptable carrier or excipient.
15. A pharmaceutical composition according to claim 1 which is capable of producing a protective immune response to N. meningitidis.