Human cytomegalovirus-specific T-cell receptor engineered for high affinity and soluble expression using mammalian cell display.

The Journal of Biological Chemistry
Ellen K WagnerJennifer A Maynard

Abstract

T-cell receptors (TCR) have considerable potential as therapeutics and antibody-like reagents to monitor disease progression and vaccine efficacy. Whereas antibodies recognize only secreted and surface-bound proteins, TCRs recognize otherwise inaccessible disease-associated intracellular proteins when they are presented as processed peptides bound to major histocompatibility complexes (pMHC). TCRs have been primarily explored for cancer therapy applications but could also target infectious diseases such as cytomegalovirus (CMV). However, TCRs are more difficult to express and engineer than antibodies, and advanced methods are needed to enable their widespread use. Here, we engineered the human CMV-specific TCR RA14 for high-affinity and robust soluble expression. To achieve this, we adapted our previously reported mammalian display system to present TCR extracellular domains and used this to screen CDR3 libraries for clones with increased pMHC affinity. After three rounds of selection, characterized clones retained peptide specificity and activation when expressed on the surface of human Jurkat T cells. We obtained high yields of soluble, monomeric protein by fusing the TCR extracellular domains to antibody hinge and Fc constan...Continue Reading

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Apr 14, 2019·The Journal of Biological Chemistry·Elissa K LeonardJamie B Spangler
Sep 10, 2020·IScience·Lucia CsepregiSai T Reddy
Feb 25, 2021·The FEBS Journal·Ross A RobinsonDavid K Cole
May 15, 2021·Journal for Immunotherapy of Cancer·Lorenzo IovinoJoshua A Hill
Dec 18, 2021·Molecular Cell·Kamyab JavanmardiIlya J Finkelstein

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