CPC C07K 14/005 (2013.01) [A61K 39/12 (2013.01); A61P 31/14 (2018.01); A61P 37/04 (2018.01); A61K 39/00 (2013.01); A61K 2039/5252 (2013.01); A61K 2039/5254 (2013.01); A61K 2039/53 (2013.01); A61K 2039/543 (2013.01); A61K 2039/55505 (2013.01); A61K 2039/55561 (2013.01); A61K 2039/55566 (2013.01); C07K 2319/21 (2013.01); C07K 2319/22 (2013.01); C07K 2319/50 (2013.01); C07K 2319/70 (2013.01); C07K 2319/735 (2013.01); C12N 2710/10343 (2013.01); C12N 2760/18522 (2013.01); C12N 2760/18523 (2013.01); C12N 2760/18534 (2013.01)] | 27 Claims |
1. A recombinant respiratory syncytial virus (RSV) F ectodomain trimer, comprising:
three recombinant F2-F1 ectodomain protomers each comprising a deletion of RSV F positions 104-144 and a glycine-serine peptide linker between RSV F positions 103 and 145, wherein the recombinant F2-F1 ectodomain protomers comprise the following amino acid substitutions to stabilize the recombinant RSV F ectodomain trimer in a prefusion conformation:
a non-native disulfide bond between cysteines introduced by 155C and 290C substitutions;
190F and 207L substitutions; and
(A) a non-native inter-protomer disulfide bond between cysteines introduced by 98C and 361C substitutions; or
(B) a non-native inter-protomer disulfide bond between cysteines introduced by 183GC and 428C substitutions, and an amino acid sequence set forth as residues 26-475 of SEQ ID NO: 2, 6, 11, 30-35, 38-39, 42-43, 46-47, 50-51, or 54-55, or residues 31-480 of SEQ ID NO: 16 or 21; and
wherein the amino acid substitutions are according to a reference RSV F sequence set forth as SEQ ID NO: 57.
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