US 12,276,656 B2
Compositions and methods for identification of antigen specific T cells
Songming Peng, San Mateo, CA (US); Boi Bryant Quach, San Mateo, CA (US); Duo An, San Mateo, CA (US); Xiaoyan Robert Bao, Foster City, CA (US); Alex Franzusoff, El Granada, CA (US); Barbara Sennino, San Francisco, CA (US); Olivier Dalmas, San Carlos, CA (US); and Stefanie Mandl-Cashman, San Francisco, CA (US)
Assigned to ADOC SSF, LLC, South San Francisco, CA (US)
Filed by ADOC SSF, LLC, South San Francisco, CA (US)
Filed on Oct. 28, 2022, as Appl. No. 18/050,715.
Application 18/050,715 is a division of application No. 16/788,745, filed on Feb. 12, 2020, granted, now 11,513,113.
Claims priority of provisional application 62/876,380, filed on Jul. 19, 2019.
Claims priority of provisional application 62/867,165, filed on Jun. 26, 2019.
Claims priority of provisional application 62/826,823, filed on Mar. 29, 2019.
Claims priority of provisional application 62/804,649, filed on Feb. 12, 2019.
Prior Publication US 2023/0194504 A1, Jun. 22, 2023
This patent is subject to a terminal disclaimer.
Int. Cl. G01N 33/50 (2006.01); A61K 39/00 (2006.01); A61P 35/00 (2006.01); C12N 5/0783 (2010.01); C12N 15/10 (2006.01); C12N 15/85 (2006.01); C12N 15/90 (2006.01); C12Q 1/686 (2018.01)
CPC G01N 33/505 (2013.01) [A61K 39/4611 (2023.05); A61K 39/4632 (2023.05); A61K 39/464401 (2023.05); A61K 39/46449 (2023.05); A61P 35/00 (2018.01); C12N 5/0636 (2013.01); C12N 5/0637 (2013.01); C12N 5/0638 (2013.01); C12N 15/1093 (2013.01); C12N 15/85 (2013.01); C12N 15/907 (2013.01); C12Q 1/686 (2013.01); A61K 2239/38 (2023.05); A61K 2239/50 (2023.05); A61K 2239/57 (2023.05); C12N 2510/00 (2013.01)] 16 Claims
 
1. A method of treating cancer in a subject, comprising:
(a) contacting a sample with a plurality of particle sets,
(i) wherein each particle of each set consists of three polypeptides comprising an antigen peptide, a barcode, and at least one identifying label;
(ii) wherein the sample comprises T cells; and
(iii) wherein contacting comprises providing conditions suitable for the T cells to bind to antigen peptides;
(b) isolating a T cell from the sample;
(c) identifying the barcodes of the particles bound to the isolated T cell;
(d) determining a ratio of a most represented barcode and a second most represented barcode identified in (c);
(e) determining the antigen specificity of the T cell based on the ratio of the most represented barcode and the second most represented barcode;
(f) identifying a TCR gene sequence of the T cell;
(g) preparing a polynucleotide comprising homology arms and at least one identified TCR gene sequence, wherein the identified TCR gene sequence is position between the homology arms;
(h) recombining the polynucleotide into an endogenous locus of the T cell;
(i) culturing the modified T cell of (h) to produce a population of T cells; and
(j) administering a therapeutically effective amount of the modified T cells to the subject, to thereby treat cancer.