Method for identifying antigen-specific t cell receptors in primate

A method comprising steps of:

• a) obtaining a population of T cells from a non-human primate that has been immunized by an antigen;
• b) preparing cDNA from the population of T cells without separating the population of T cells into single cells;
• c) sequencing the cDNA to obtain:
• (c1) a plurality of T-cell receptor (TCR) α-chain sequences and
• (c2) a plurality of TCR β-chain sequences,
• wherein each of the TM α-chain sequences comprises an α-chain framework, CDR-α1, CDR-α2 and CDR-α3, and wherein each of the TCR β-chain sequences comprises a β-chain framework, CDR-β1, CDR-β2 and CDR-β3;
• d) grouping said obtained plurality of TCR α-chain sequences into a plurality of α-chain groups, wherein the CDR-α3 of said α-chain sequences in each α-chain group contain 0, 1 or 2 amino acid differences relative to one another;
• e) grouping said obtained plurality of TCRβ-chain sequences into a plurality of β-chain groups, wherein the CDR-β3 of said β-chain sequences in each β-chain group contain 0, 1 or 2 amino acid differences relative to one another;
• f) ranking the plurality of α-chain groups according to the number of TCR β-chain sequences in each α-chain group, wherein
• a first α-chain group has the most members of TCR α-chain sequences and a
• second α-chain group has the second most members of TCR α-chain sequences,
• and so on-for the plurality of α-chain groups;
• g) ranking the plurality of β-chain groups according to the number of TCR β-chain sequences in each β-chain group, wherein
• a first β-chain group has the most members of TCR β-chain sequences and a
• second β-chain group has the second most members of TCR β-chain sequences,
• and so on for the plurality of β-chain groups;
• h) pairing the first α-chain group with the first β-chain group;
• i) pairing a first TCR α-chain sequence from the first α-chain group and a first TCR β-chain sequence from the first β-chain group,
• wherein
• the first TCR α-chain sequence comprises a first α-chain framework, first CDR-α1, first CDR-α2 and first CDR-α3, and
• the first TCR β-chain sequence comprises a first β-chain framework, first CDR-β1, first CDR-β2 and first CDR-β3, and
• j) testing a first candidate TCR comprising the first TCR α-chain sequence from the first α-chain group and the first TCR β-chain sequence from the first β-chain group for binding to the antigen.