Modified cytomegalovirus proteins and stabilized complexes
AssigneeGlaxosmithkline Biologicals SA
InventorsEnrico MALITO, Matthew James BOTTOMLEY, Andrea CARFI, Sumana CHANDRAMOULI, Kate LUISI
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Abstract
Described are mutant human cytomegalovirus (HCMV) pentamer complex polypeptides, methods of making them, and their use in HCMV protein complexes and compositions. In particular, the use of the modified HCMV polypeptides to stabilize HCMV complexes or unmask a pentamer epitope is described.
Claims12
1-18. (canceled)
A Human Cytomegalovirus (HCMV) glycoprotein H (gH) polypeptide and/or complex-forming fragment thereof comprising a stabilizing mutation, wherein:
- (a) the stabilizing mutation comprises a cavity-filling mutation, a hydrophobic mutation, a hydrophilic mutation, a disulphide bridge mutation, or a deglycosylation mutation, and
- (b) the HCMV gH polypeptide and/or complex-forming fragment thereof comprises A102, A372, A352, or L257, each of which are as in SEQ ID NO:
- 1 or at a corresponding position, substituted with a tryptophan (W), phenylalanine (F), tyrosine (Y), or leucine (L).
An HCMV gH polypeptide comprising:
- (a) an amino acid sequence that is at least 90% identical to SEQ ID NO:
- 1 and
- (b) at least one of (i)-(v):
- (i) at least one of A102, A372, A352, and L257 independently substituted with tryptophan (W), phenylalanine (F), tyrosine (Y), or leucine (L);
- (ii) at least one of H252, K404, R255, E355, H480, S601, and R405 independently substituted with tryptophan (W), phenylalanine (F), methionine (M), cysteine (C), alanine (A), leucine (L), isoleucine (I), valine (V), or proline (P);
- (iii) at least one of G358 and H327 independently substituted with serine (S), threonine (T), cysteine (C), tyrosine (Y), asparagine (N), glutamine (Q), arginine (R), glutamic acid (E), lysine (K), histidine (H), or aspartic acid (D);
- (iv) at least one of V109 and LIII independently substituted with a cysteine (C);
- and
- (v) at least one of N55, N62, N67, N192, N641, and N700 independently substituted with glutamine (Q), serine (S), threonine (T), alanine (A), glutamate (E), or aspartate (D).
A nucleic acid molecule encoding an HCMV gH polypeptide and/or complex-forming fragment thereof comprising a stabilizing mutation, wherein:
- (a) the stabilizing mutation comprises a cavity-filling mutation, a hydrophobic mutation, a hydrophilic mutation, a disulphide bridge mutation, or a deglycosylation mutation, and
- (b) the HCMV gH polypeptide or complex-forming fragment thereof comprises A102, A372, A352, or L257, each of which are as in SEQ ID NO:
- 1 or at a corresponding position, substituted with a tryptophan (W), phenylalanine (F), tyrosine (Y), or leucine (L).
Dependencies are inferred from the claim text.