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Synthesis and Bioconjugation of Thiol-Reactive Reagents for the Creation of Site-Selectively Modified Immunoconjugates
Published on: March 6, 2019
Antibody Conjugation to Cell Surface Receptors
Avani Parikh1,2,3, Elisa Ruffo1,2,3, Alexander Deiters1,4
1UPMC Hillman Cancer Center, University of Pittsburgh, Pittsburgh, PA, USA.
Methods in Molecular Biology (Clifton, N.J.)
|August 1, 2026
Summary
Researchers developed SNAP-CAR, a novel universal chimeric antigen receptor (CAR), for precise T cell targeting. This method covalently links CARs to antibodies, enabling programmable T cell signaling for advanced cell therapies and research tools.
Area of Science:
- Immunology
- Biotechnology
- Molecular Biology
Background:
- Universal chimeric antigen receptors (CARs) offer programmable T cell signaling.
- Current methods require specific CARs for each target antigen.
Purpose of the Study:
- To describe a method for covalent cell surface modification using SNAP-tag enzymes and antibodies.
- To introduce SNAP-CAR, a universal CAR system for adaptable T cell targeting.
- To provide protocols for generating SNAP-CAR T cells and adaptor antibodies.
Main Methods:
- Bioconjugation of antibodies to SNAP-tag enzyme on T cells.
- Gamma retroviral transduction for SNAP-CAR T cell generation.
- Benzyl guanine (BG)-NHS ester conjugation for antibody adaptors.
- Quantification of BG motifs and assessment of cell surface bioconjugate formation.
- Assays for antigen-directed T cell functions: target cell killing and activation.
Main Results:
- Successful covalent attachment of SNAP-CAR to adaptor antibodies via BG motif.
- Demonstrated protocols for generating modified T cells and antibodies.
- Validated methods for evaluating bioconjugate formation and function.
- Showcased antigen-directed T cell killing and activation mediated by SNAP-CAR.
Conclusions:
- SNAP-CAR enables precise, covalent cell surface modification for programmable T cell signaling.
- This universal CAR approach enhances flexibility for cell therapeutics and research.
- The described protocols facilitate the generation and validation of SNAP-CAR systems.
Keywords:
Antibody conjugationAntibody therapeuticsCAR T cell therapySynthetic biologyUniversal CAR T cellsMore Related Videos
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