Generation of higher affinity T cell receptors by antigen-driven differentiation of progenitor t cells in vitro

Thomas M. Schmitt, David H. Aggen, Kumiko Ishida-Tsubota, Sebastian Ochsenreither, David M. Kranz, Philip D. Greenberg

Research output: Contribution to journalArticlepeer-review


Many promising targets for T-cell-based cancer immunotherapies are self-antigens. During thymic selection, T cells bearing T cell receptors (TCRs) with high affinity for self-antigen are eliminated. The affinity of the remaining low-avidity TCRs can be improved to increase their antitumor efficacy, but conventional saturation mutagenesis approaches are labor intensive, and the resulting TCRs may be cross-reactive. Here we describe the in vitro maturation and selection of mouse and human T cells on antigen-expressing feeder cells to develop higher-affinity TCRs. The approach takes advantage of natural Tcrb gene rearrangement to generate diversity in the length and composition of CDR3b. In vitro differentiation of progenitors transduced with a known Tcra gene in the presence of antigen drives differentiation of cells with a distinct agonist-selected phenotype. We purified these cells to generate TCRb chain libraries pre-enriched for target antigen specificity. Several TCRb chains paired with a transgenic TCRa chain to produce a TCR with higher affinity than the parental TCR for target antigen, without evidence of cross-reactivity.

Original languageEnglish (US)
Pages (from-to)1188-1195
Number of pages8
JournalNature Biotechnology
Issue number12
StatePublished - 2017

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Molecular Medicine
  • Biomedical Engineering


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