Authors
Avani Parikh, Elisa Ruffo, Alexander Deiters, Jason Lohmueller
Published in
Methods in molecular biology (Clifton, N.J.). Volume 3009. Pages 129-151.
Abstract
Here, we describe the covalent modification of cell surfaces with antibodies through bioconjugation to the self-labeling SNAP-tag enzyme. We apply this approach to the reprogramming of T cell signaling through the universal chimeric antigen receptor (CAR), "SNAP-CAR." Universal CARs are a highly programmable class of engineered receptors that interact with co-administered "adaptor" antibodies to recognize one or more antigens of interest to trigger receptor signaling. Universal CARs have promise for use in cell therapeutics and as research tools. SNAP-CAR covalently attaches to adaptor antibodies containing a benzyl guanine (BG) motif. The protocols presented here include methods for SNAP-CAR T cell and antibody adaptor generation using gamma retroviral transduction and BG-NHS ester conjugation, respectively. The chapter also describes methods to evaluate cell surface bioconjugate formation, including quantification of BG motifs on antibodies and co-incubation experiments to assay for antigen-directed SNAP-CAR T cell functions of target cell killing and SNAP-CAR T cell activation.
PMID:
42542485
Bibliographic data and abstract were imported from PubMed on 02 Aug 2026.
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