	US 12,486,501 B2
	Multiplexed testing of lymphocytes for antigen specificity
David Gifford, Newton, MA (US); and Brandon Carter, Cambridge, MA (US)
Assigned to Think Therapeutics, Inc., Newton, MA (US)
Filed by Think Therapeutics, Inc., Newton, MA (US)
Filed on Jul. 28, 2021, as Appl. No. 17/386,702.
Application 17/386,702 is a continuation of application No. 17/142,745, filed on Jan. 6, 2021, granted, now 11,111,489.
Prior Publication US 2022/0213466 A1, Jul. 7, 2022
Int. Cl. C12N 15/10 (2006.01); C40B 30/04 (2006.01)

CPC C12N 15/1037 (2013.01) [C12N 15/1089 (2013.01); C40B 30/04 (2013.01)]

15 Claims

1. A method for determining a T cell receptor chain sequence, or a portion thereof, specific for two or more antigens, the method comprising:

sorting a plurality of first antigens into a plurality of reaction mixtures, wherein the sorting comprises adding two or more unique antigens of the plurality of first antigens to two or more unique subsets of the plurality of reaction mixtures, and wherein the two or more unique antigens are not added to any two identical subsets of the plurality of reaction mixtures;

contacting each reaction mixture with a biological sample comprising a plurality of T cells;

providing a condition for a first activated T cell in at least one reaction mixture of the plurality of reaction mixtures to expand in number such that a plurality of T cell clones is formed;

adding a query antigen to at least one unique subset of the two or more unique subsets of the plurality of reaction mixtures;

separating a second activated T cell and a non-activated T cell from a subset of the plurality of reaction mixtures, wherein the second activated T cell recognizes the query antigen;

sequencing nucleic acids of the second activated T cell to obtain the T cell receptor chain sequence; and

detecting the two or more unique antigens of the plurality of first antigens, wherein the two or more unique antigens are specific for the T cell receptor chain sequence.