@inbook{fae304e82d06450a8c44c7422760b653,
title = "T cell receptor engineering",
abstract = "T lymphocytes express on their surface a heterodimeric αβ receptor, called the T cell receptor (TCR), which recognizes foreign antigens. Unlike antibodies, the recognition requires both an antigenic peptide epitope and a protein encoded by the major histocompatibility complex (MHC). In contrast to conventional antibody-directed target antigens, antigens recognized by the TCR can include the entire array of potential intracellular proteins, which are processed and delivered to the cell surface as a peptide/MHC complex. In the past 10 years, there have been significant efforts to engineer TCRs in various formats, which would allow improved recognition and destruction of virus-infected cells or cancer. The proposed therapeutic approaches involve either the use of engineered, high-affinity TCRs in soluble forms, analogous to antibody-directed therapies, or the use of engineered TCRs whose genes are reintroduced into autologous T cells and transferred back into patients (T cell adoptive therapies). This chapter describes three methods associated with the engineering of TCRs for these therapeutic purposes: (1) use of a yeast display system to engineer higher affinity single-chain VαVβ TCRs, called scTv; (2) use of a T cell display system to engineer higher affinity full-length TCRs; and (3) expression, purification, and characterization of soluble TCRs in an Escherichia coli system.",
keywords = "Complementarity-determining regions, Fluorescence-activated cell sorting, Keywords, Major histocompatibility complex, Protein engineering, Splicing by overlap extension, T cell display, T cell receptor, Yeast display",
author = "Stone, {Jennifer D.} and Chervin, {Adam S.} and Aggen, {David H.} and Kranz, {David M.}",
note = "Funding Information: We thank members of the lab, past and present, for valuable contributions to the development of these methods. Research in the lab has been funded by grants from the National Institutes of Health R01 GM55767, R01 AI064611, P01 CA97296, and U54 AI57153 (from the NIH-supported Great Lakes Regional Center for Excellence) to D. M. K. and a Samuel and Ruth Engelberg/Irvington Institute Postdoctoral Fellowship of the Cancer Research Institute to J. D. S.",
year = "2012",
doi = "10.1016/B978-0-12-396962-0.00008-2",
language = "English (US)",
series = "Methods in Enzymology",
publisher = "Academic Press Inc.",
pages = "189--222",
booktitle = "Methods in Enzymology",
address = "United States",
}