Patent ID: 11913024
Assignee: JUNO THERAPEUTICS, INC.
Field: Pharmaceuticals (Chemistry)
Classification: CPC C  A | IPC C

Claim 0:
1. A method for stimulating target T cells, the method comprising:
(a) adding a composition comprising target T cells to a chromatography matrix present in a chromatography column, wherein a selection agent is immobilized on the chromatography matrix, the selection agent comprising an antibody or antibody fragment that specifically binds to a selection marker expressed by the target T cells, wherein specific binding by the antibody or antibody fragment of the selection agent to the selection marker effects the reversible immobilization of at least a plurality of the target T cells on the chromatography matrix;
(b) adding a stimulatory reagent to the chromatography matrix, wherein the stimulatory reagent is in soluble form and comprises a first stimulatory agent, a second stimulatory agent, and an oligomer or polymer of at least 50 crosslinked tetramers of a streptavidin mutein, wherein:
the first stimulatory agent comprises an anti-CD3 antibody or antibody fragment and a first streptavidin-binding peptide that is reversibly bound to the oligomer or polymer; and
the second stimulatory agent comprises an anti-CD28 antibody or antibody fragment and a second streptavidin-binding peptide that is reversibly bound to the oligomer or polymer; and
(c) incubating the at least a plurality of immobilized target T cells in the presence of the stimulatory reagent in the chromatography column, wherein
the incubation effects the stimulation of T cells of the at least a plurality of immobilized target T cells and is carried out under conditions whereby (i) the anti-CD3 antibody or antibody fragment specifically binds to a CD3 molecule expressed on the surface of the T cells, thereby inducing a primary activation signal in the T cells, and (ii) the anti-CD28 antibody or antibody fragment specifically binds to a CD28 molecule expressed on the surface of the T cells, thereby inducing a costimulatory signal in the T cells.