	US 12,129,465 B2
	Stable coronavirus proteins and vaccine compositions thereof
Daniel Ellis, Seattle, WA (US); Neil King, Seattle, WA (US); Jesse Bloom, Seattle, WA (US); Tyler Starr, Seattle, WA (US); and Allison Greaney, Seattle, WA (US)
Assigned to UNIVERSITY OF WASHINGTON, Seattle, WA (US); and FRED HUTCHINSON RESEARCH CENTER, Seattle, WA (US)
Filed by UNIVERSITY OF WASHINGTON, Seattle, WA (US); and FRED HUTCHINSON CANCER RESEARCH CENTER, Seattle, WA (US)
Filed on Dec. 28, 2021, as Appl. No. 17/563,271.
Claims priority of provisional application 63/210,654, filed on Jun. 15, 2021.
Claims priority of provisional application 63/188,651, filed on May 14, 2021.
Claims priority of provisional application 63/132,863, filed on Dec. 31, 2020.
Prior Publication US 2022/0325279 A1, Oct. 13, 2022
Int. Cl. C07K 14/165 (2006.01); A61K 39/215 (2006.01); C12N 15/11 (2006.01); C12N 15/64 (2006.01); C12Q 1/6811 (2018.01)

CPC C12N 15/11 (2013.01) [A61K 39/215 (2013.01); C07K 14/165 (2013.01); C12N 15/64 (2013.01); C12Q 1/6811 (2013.01)]

31 Claims

1. A non-naturally occurring polypeptide comprising:

a coronavirus receptor binding domain (RBD) comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of residues 328-531 of SEQ ID NO:1 and further comprising at least two mutations relative to SEQ ID NO: 1,

wherein the polypeptide exhibits a melting temperature measured by nano differential scanning fluorimetry at least 2° C. higher than a polypeptide comprising residues 328-531 of SEQ ID NO: 1 and lacking the at least two mutations, and

wherein the at least two mutations are selected from the group consisting of:

F338L and Y365W;

Y365W and L513M;

Y365W and F392W;

F338M and A363L and Y365F and F377V;

Y365F and F392W;

Y365F and V395I;

Y365F and F392W and V395I;

Y365W and L513I and F515L;

F338L and A363L and Y365M;

F338L and I358F and Y365W;

I358F and Y365W and L513M;

I358F and Y365W and F392W;

I358F and Y365F and V395I; and

I358F and Y365F and F392W and V395I.