| CPC C07K 14/70539 (2013.01) [C07K 14/61 (2013.01); C12N 15/1068 (2013.01); G01N 33/56972 (2013.01); C07K 2319/02 (2013.01); C07K 2319/20 (2013.01); C07K 2319/21 (2013.01); C07K 2319/50 (2013.01)] | 17 Claims |

|
1. A method for isolating an antigen-specific T cell, the method comprising:
a. preparing a plurality of particles by
i) obtaining a library comprising at least two polynucleotide, each polynucleotide comprising, from 5′ to 3′, a sequence insert comprising a stop codon in each of the three reading frames of the polynucleotide, a Beta 2 Microglobulin (β2M) coding sequence, and a Major Histocompatibility Complex (MHC) heavy chain coding sequence, wherein the sequence insert is flanked at its 5′ or 3′ by a first universal target sequence comprising a restriction site or a polymerase chain reaction (PCR) primer target site;
ii) inserting a polynucleotide encoding an antigenic sequence into the sequence insert, to thereby obtain a polypeptide comprising, in an amino to carboxyl terminus orientation, the antigenic sequence, the Beta 2 Microglobulin (β2M) sequence, and the Major Histocompatibility Complex (MHC) heavy chain sequence;
iii) attaching the polypeptide to a particle to obtain a plurality of particles;
b. contacting the plurality of particles with a plurality of T cells under conditions suitable for antigen-specific binding of a T cell to a particle; and
c. isolating the particles, to thereby isolate an antigen-specific T cell bound thereto;
wherein the particle is a magnetic bead, an agarose bead, a styrene polymer particle, or a dextran polymer particle.
|