US 12,304,928 B2
Epitope restriction for antibody selection
Payton Weidenbacher, Stanford, CA (US); and Peter S. Kim, Stanford, CA (US)
Assigned to CZ Biohub SF, LLC, San Francisco, CA (US); and The Board of Trustees of the Leland Stanford Junior University, Stanford, CA (US)
Appl. No. 17/053,230
Filed by CZ Biohub SF LLC, San Francisco, CA (US); and The Board of Trustees of the Leland Stanford Junior University, Stanford, CA (US)
PCT Filed May 17, 2019, PCT No. PCT/US2019/032945
§ 371(c)(1), (2) Date Nov. 5, 2020,
PCT Pub. No. WO2019/222674, PCT Pub. Date Nov. 21, 2019.
Claims priority of provisional application 62/780,595, filed on Dec. 17, 2018.
Claims priority of provisional application 62/688,939, filed on Jun. 22, 2018.
Claims priority of provisional application 62/673,617, filed on May 18, 2018.
Prior Publication US 2021/0139542 A1, May 13, 2021
Int. Cl. A61K 39/00 (2006.01); C07K 1/107 (2006.01); C07K 14/005 (2006.01); C07K 16/10 (2006.01)
CPC C07K 14/005 (2013.01) [A61K 39/00 (2013.01); C07K 1/1072 (2013.01); C07K 16/1018 (2013.01); C07K 16/1063 (2013.01); C07K 2317/35 (2013.01); C12N 2740/16122 (2013.01); C12N 2760/16122 (2013.01)] 13 Claims
 
1. A method of making a modified hemagglutinin influenza virus protein, or a fragment or a variant thereof, the method comprising the steps of:
(a) providing a hemagglutinin influenza virus protein, or the fragment or the variant thereof, comprising a target region comprising an epitope of interest and a non-target region;
(b) providing a binding partner, wherein the binding partner binds specifically to the target region;
(c) contacting the hemagglutinin influenza virus protein, or the fragment or the variant thereof, with the binding partner under conditions in which the binding partner binds specifically to the target region, thereby forming a protein complex comprising the hemagglutinin influenza virus protein, or the fragment or the variant thereof, and the binding partner;
(d) contacting amino acid residues of the hemagglutinin influenza virus protein, or the fragment or the variant thereof, in the protein complex with a modifying reagent under conditions sufficient to form the modified hemagglutinin influenza virus protein, or the fragment or the variant thereof, thereby forming the modified hemagglutinin influenza virus protein, or the fragment or the variant thereof, in the protein complex, wherein each of a plurality of amino acid residues in the non-target region of the hemagglutinin influenza virus protein, or the fragment or the variant thereof, have a modifying component covalently attached thereto, and wherein amino acid residues of the target region of the hemagglutinin influenza virus protein, or the fragment or the variant thereof, do not have the modifying component attached thereto; and
(e) separating the modified hemagglutinin influenza virus protein, or the fragment or the variant thereof, from the binding partner in the protein complex, thereby providing the modified hemagglutinin influenza virus protein, or the fragment or the variant thereof, with reduced or ablated immunogenicity of the non-target region.