US 11,746,143 B2
Methods to identify immunogens by targeting improbable mutations
Barton F. Haynes, Durham, NC (US); Kevin J. Wiehe, Durham, NC (US); and Mattia Bonsignori, Durham, NC (US)
Assigned to DUKE UNIVERSITY, Durham, NC (US)
Filed by Duke University, Durham, NC (US)
Filed on Oct. 13, 2021, as Appl. No. 17/500,750.
Application 17/500,750 is a continuation of application No. 16/337,264, granted, now 11,161,895, previously published as PCT/US2017/054956, filed on Oct. 3, 2017.
Application 16/337,264 is a continuation in part of application No. PCT/US2017/020823, filed on Mar. 3, 2017.
Claims priority of provisional application 62/489,250, filed on Apr. 24, 2017.
Claims priority of provisional application 62/476,985, filed on Mar. 27, 2017.
Claims priority of provisional application 62/403,649, filed on Oct. 3, 2016.
Claims priority of provisional application 62/403,635, filed on Oct. 3, 2016.
Prior Publication US 2022/0185871 A1, Jun. 16, 2022
Int. Cl. C12N 7/00 (2006.01); A61K 39/00 (2006.01); A61K 39/12 (2006.01); A61K 51/10 (2006.01); C07K 14/005 (2006.01); C07K 16/10 (2006.01); A61K 39/21 (2006.01); G01N 33/569 (2006.01)
CPC C07K 16/1063 (2013.01) [A61K 39/21 (2013.01); C12N 7/00 (2013.01); G01N 33/56983 (2013.01); C07K 2317/55 (2013.01); C07K 2317/56 (2013.01); C07K 2317/92 (2013.01); C12N 2740/16034 (2013.01)] 36 Claims
 
1. A method for identifying improbable mutations in the heavy or light chains of broadly neutralizing anti-viral-antigen antibodies comprising:
(a) identifying at least one improbable somatic mutation in the heavy or light chain variable domain of a broadly neutralizing anti-viral-antigen antibody compared to the sequence of the corresponding unmutated common ancestor (UCA) antibody, wherein the somatic mutation is an improbable somatic mutation if in the absence of antigenic selection the somatic mutation occurs in the broad-neutralizing anti-viral-antigen antibody with a probability of less than 2%;
(b) reverting the at least one improbable somatic mutation identified in step (a) to its UCA encoded amino acid(s) to thereby provide a recombinant antibody;
(c) expressing the recombinant antibody of step (b) and testing the expressed recombinant antibody for neutralizing activity against a virus that comprises the viral antigen or for binding ability against the viral antigen, and
(d) determining whether the improbable mutation identified in step (a) is functionally significant by testing whether the expressed recombinant antibody of step (c) exhibits a reduction of neutralizing activity or reduction of binding ability as compared to an antibody with the same amino acid sequence but for the reverted amino acid sequence.