US 12,215,313 B2
Method of detecting new immunogenic T cell epitopes and isolating new antigen-specific T cell receptors by means of an MHC cell library
Felix Lorenz, Berlin (DE); Wolfgang Uckert, Berlin (DE); Christian Ellinger, Munich (DE); and Dolores Schendel, Munich (DE)
Assigned to MAX-DELBRÜCK-CENTRUM FÜR MOLEKULARE MEDIZIN IN DER HELMHOLTZ-GEMEINSCHAFT, Berlin (DE); and ZENTRUM MÜNCHEN-DEUTSCHES FORSCHUNGSZENTRUM FÜR GESUNDHEIT UND UMWELT (GMBH), Neuherberg (DE)
Filed by MAX-DELBRÜCK-CENTRUM FÜR MOLEKULARE MEDIZIN IN DER HELMHOLTZ-GEMEINSCHAFT, Berlin (DE); and HELMHOLTZ ZENTRUM MÜNCHEN-DEUTSCHES FORSCHUNGSZENTRUM FÜR GESUNDHEIT UND UMWELT (GMBH), Neuherberg (DE)
Filed on May 6, 2021, as Appl. No. 17/313,761.
Application 17/313,761 is a division of application No. 15/558,765, granted, now 11,001,830, previously published as PCT/EP2016/055518, filed on Mar. 15, 2016.
Claims priority of application No. 15159212 (EP), filed on Mar. 16, 2015.
Prior Publication US 2021/0261947 A1, Aug. 26, 2021
Int. Cl. C12N 15/10 (2006.01); C07K 14/005 (2006.01); C07K 14/725 (2006.01)
CPC C12N 15/1037 (2013.01) [C07K 14/005 (2013.01); C07K 14/7051 (2013.01); C12N 2710/16122 (2013.01); C12N 2710/16134 (2013.01); C12N 2710/20022 (2013.01); C12N 2710/20034 (2013.01)] 8 Claims
 
1. A method for preparing a nucleic acid encoding the TRA and TRB of a TCR construct specific for an immunodominant epitope from a defined antigen presented on a MHC, comprising
(a) stimulating T cells isolated from a donor with professional antigen presenting cells presenting epitopes of said defined antigen, to enrich antigen-specific T cells; and
(b) contacting said T cells with a library of cells, wherein each cell expresses a single MHC allele, wherein the library comprises cells expressing all MHC II alleles present in the donor, and wherein the cells of said library present epitopes of said defined antigen; wherein T cells expressing TCR specific for said epitopes become activated T cells; and
(c) selecting T cells activated by said contact in step b, preferably, based on an activation marker expressed by said activated T cells; and
(d) isolating the nucleic acids encoding the TCR alpha and TCR beta chains of the TCR of said activated T cells selected in step c.