Neutralising antibody against dengue for use in a method of prevention and/or treatment of Zika infection
A flavivirus Envelope Dimer Epitope (EDE) and isolated neutralizing antibody or antigen binding fragment thereof directed against the EDE for use in vaccinating an individual against one or more flaviviruses wherein the EDE is a stabilized recombinant flavivirus are provided. The dimer is: covalently stabilized with at least one disulphide inter-chain bond or one sulfhydryl-reactive crosslinker between the two sE monomers, and/or by being formed as a single polypeptide chain, and/or by linking the two sE monomers through modified sugar, and/or non-covalently stabilized by substituting at least one amino acid residue in the amino acid sequence of at least one sE monomer with at least one bulky side chain amino acid, at the dimer interface or in domain 1 (D1)/domain 3 (D3) linker of each monomer. The dimer is a homodimer or heterodimer of native and/or mutant envelope polypeptides, from DENV-1, DENV-2, DENV-3, DENV-4, Zika and/or other flavivirus.
1. A composition comprising an isolated neutralizing antibody or antigen binding fragment thereof directed against a flavivirus envelope dimer epitope (EDE) and an adjuvant,
wherein the antibody or fragment comprises six CDRs wherein each CDR comprises an amino acid sequence selected from the group consisting of:
SEQ ID Nos: 15 to 26 and sequences with no more than 30% modification from any one of said SEQ ID No. 15 to 26,
wherein:
(a) said antibody or fragment thereof binds the five polypeptide segments of the dengue virus glycoprotein E ectodomain (sE) consisting of the residues 67-74, residues 97-106, residues 307-314, residues 148-159 and residues 243-251, or corresponding residues of the flavivirus or Zika virus glycoprotein E ectodomain, or consisting of Zika PF13 residues 67-77, residues 97-106, residues 313-315, residues 243-253, residue K373 or corresponding residues of the flavivirus glycoprotein E ectodomain, or
(b) binding is unaffected by presence or absence of dengue N153 (Zika N154) glycan or corresponding residue.
2. The composition of claim 1 , wherein the antibody or fragment thereof recognizes exclusively virion-dependent (including sub-viral particle or virus-like particle) epitope(s) of a flavivirus, optionally Zika or dengue virus.
3. The composition of claim 1 , wherein the fragment is a Fab fragment.
4. The composition of claim 1 , wherein the antibody or fragment thereof comprises an amino acid sequence selected from the group consisting of SEQ ID Nos: 15 to 26 and sequences with no more than 20, 15, or 10% modification from any one of said SEQ ID No. 15 to 26.
5. The composition of claim 1 wherein the EDE comprises a region centred in a valley lined by the h strand on the domain II side, and the “150 loop” on the domain I side (across from the dimer interface), wherein the 150 loop spans residues 148-159, connecting b-strands E0 and F0 of domain 1, and carries the N153 glycan, which covers the fusion loop of the partner subunit in the dimer, optionally wherein the region comprises the h strand (residues 67-74 which bear the N67 glycan), the fusion loop and residues immediately upstream (residues 97-106) and the ij loop (residues 246-249) of the reference subunit, wherein the reference subunit is the subunit which contributes the fusion loop,
optionally wherein the EDE further comprises the 150 loop and the N153 glycan chain of the second subunit,
optionally wherein one or both regions is in a similar spatial configuration as the native region;
or wherein the EDE comprises the Zika PF13 beta strand b of domain IL bed beta-sheet edge, fusion loop main chain, fusion loop R99 side chain, Q77 side chain, disulphide bond between C74 and C105; beta strand E, K373, charged residues in domain I, kl loop of domain II, or regions corresponding thereto.
6. The composition of claim 1 , wherein the antibody or fragment thereof neutralises one or more serotypes of Dengue virus and/or Zika virus to 80%.
7. The composition of claim 1 , wherein the antibody or fragment thereof neutralises all serotypes of Dengue virus and Zika virus, to 80%.
8. The composition of claim 1 wherein the antibody or fragment thereof neutralises one or more serotypes of Dengue virus and/or Zika virus to 80, 90, 98 or 100% at a concentration of 0.5-0.01 μg/ml.
9. The compositional of claim 1 wherein the antibody or fragment thereof neutralises all serotypes of Dengue virus and Zika s to 80, 90, 98 or 100% at a concentration of 0.5-0.01 μg/ml.
10. The composition of claim 1 wherein the fragment is a Fv fragment; a Fab-like fragment; or a domain antibody; or wherein the antibody is a monoclonal antibody or a recombinant antibody.
11. The composition of claim 1 wherein the antibody is a polyclonal antibody or antigen binding portion thereof.
12. The composition of claim 1 wherein the antibody or fragment thereof is part of a composition comprising a mixture of antibodies, optionally:
a) a mixture of monoclonal antibodies or antigen binding portion thereof, or
b) a mixture of polyclonal antibodies or antigen binding portion thereof, or
c) a mixture or monoclonal and polyclonal antibodies or antigen binding portion thereof.
13. The composition of claim 6 , wherein the antibody or fragment thereof neutralises one or more serotypes of Dengue virus and/or Zika virus to 90% or 98% or 100%.
14. The composition of claim 13 , wherein the antibody or fragment thereof neutralises one or more serotypes of Dengue virus and/or Zika virus to 98% or 100%.
15. The composition of claim 7 , wherein the antibody or fragment thereof neutralises all serotypes of Dengue virus and Zika virus to 90%.
16. The composition of claim 7 , wherein the antibody or fragment thereof neutralises all serotypes of Dengue virus and Zika virus, 98% or 100%.
17. The composition of claim 16 , wherein the antibody or fragment thereof neutralises all serotypes of Dengue virus and Zika virus to 100% at the same concentration of antibody or fragment.
18. The composition of claim 10 wherein the Fab-like fragment is a Fab′ fragment or a F(ab)2 fragment.
19. A composition comprising an adjuvant and an isolated neutralizing antibody or antigen binding fragment thereof directed against a flavivirus envelope dimer epitope (EDE), wherein the antibody or fragment comprises a heavy chain comprising SEQ ID NO: 120 and/or a light chain comprising SEQ ID NO: 121, or a sequence with at least 90% homology to the said sequences.