T cell receptor engineering.
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.
Department of Biochemistry, University of Illinois, Urbana, Illinois, USA.
Jennifer D
JD
Stone
Adam S
AS
Chervin
David H
DH
Aggen
David M
DM
Kranz
D003001:Cloning, Molecular; D022801:Complementarity Determining Regions / Q000737:chemistry / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000737:chemistry; D004926:Escherichia coli / Q000737:chemistry; D005434:Flow Cytometry; D006801:Humans; D008285:Major Histocompatibility Complex; D019151:Peptide Library; D015202:Protein Engineering / Q000379:methods*; D058849:Protein Refolding; D055550:Protein Stability; D011948:Receptors, Antigen, T-Cell / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000627:therapeutic use; D011956:Receptors, Cell Surface; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D057127:Single-Chain Antibodies / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000627:therapeutic use; D012995:Solubility; D013601:T-Lymphocytes / Q000737:chemistry*; D019076:Transgenes; D015003:Yeasts / Q000737:chemistry
D022801:Complementarity Determining Regions; D000939:Epitopes; D019151:Peptide Library; D011948:Receptors, Antigen, T-Cell; D011956:Receptors, Cell Surface; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies