US 11,878,998 B2
Prefusion RSV F proteins and their use
Peter Kwong, Washington, DC (US); Barney Graham, Rockville, MD (US); John Mascola, Rockville, MD (US); Li Ou, Potomac, MD (US); Aliaksandr Druz, Germantown, MD (US); Man Chen, Bethesda, MD (US); Wing-Pui Kong, Germantown, MD (US); Ivelin Stefanov Georgiev, Nashville, TN (US); Emily Rundlet, New York, NY (US); Michael Gordon Joyce, Washington, DC (US); Yaroslav Tsybovsky, Frederick, MD (US); Paul Thomas, Washington, DC (US); Marie Pancera, Seattle, WA (US); Mallika Sastry, Rockville, MD (US); Cinque Soto, Nashville, TN (US); Joseph Van Galen, North Wales, PA (US); Guillaume Stewart-Jones, Cambridge, MA (US); Yongping Yang, Potomac, MD (US); Baoshan Zhang, Bethesda, MD (US); and Ulrich Baxa, Frederick, MD (US)
Assigned to The United States of America, as represented by the Secretary Department of Health and Human Services, Bethesda, MD (US)
Filed by The United States of America, as represented by the Secretary, Department of Health and Human Services, Bethesda, MD (US)
Filed on Nov. 11, 2021, as Appl. No. 17/524,380.
Application 17/524,380 is a continuation of application No. 16/089,993, granted, now 11,174,292, previously published as PCT/US2017/024714, filed on Mar. 29, 2017.
Claims priority of provisional application 62/314,946, filed on Mar. 29, 2016.
Prior Publication US 2022/0064222 A1, Mar. 3, 2022
Int. Cl. C07K 14/005 (2006.01); A61K 39/12 (2006.01); A61P 37/04 (2006.01); A61P 31/14 (2006.01); A61K 39/00 (2006.01)
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.