Anti-CAR Whitlow Linker [1C3C3]

The anti-CAR Whitlow linker peptide 1C3C3 clone has been developed to detect cells expressing Whitlow linker-containing CARs with different antigen specificities, including those harbouring the widely employed anti-CD19 FMC63-derived scFv as well as other scFvs, such as those targeting B-cell maturation antigen (BCMA) or CD33. It has been demonstrated to have a mean KD of […]

Contributors

Inventor Institute
Erik Kimble, Jocelyn Wright Fred Hutchinson Cancer Center
Cat. #: 162441
Unit size: 1 mg
Application: Flow cytometry, IHC, stimulation of T cells
Target: Chimeric antigen receptor Whitlow scFv peptide linker
Clone: 1C3C3
Host: Mouse
Class: Monoclonal
Primary citation: Kimble et al. 2025.J Immunother Cancer. 2025 Nov 18;13(11):e013123. PMID: 41253500
Product description: The anti-CAR Whitlow linker peptide 1C3C3 clone has been developed to detect cells expressing Whitlow linker-containing CARs with different antigen specificities, including those harbouring the widely employed anti-CD19 FMC63-derived scFv as well as other scFvs, such as those targeting B-cell maturation antigen (BCMA) or CD33. It has been demonstrated to have a mean KD of 1 to 2.6 µM when tested for binding kinetics against FMC63-Whitlow scFv protein and the Whitlow peptide. No binding to G4S4 peptide was found. In addition, the 1C3C3 antibody has been demonstrated to stain engineered T cells expressing 1H7- or FMC63-Whiltow, but not 1H7- or FMC63- G4S4 by FACS analysis. In functional assays, 1C3C3 acts as a potent agonist antibody: when immobilized, it robustly activates Whitlow‑linker CAR‑expressing Jurkat reporter cells and primary CAR T cells, driving CAR signalling, cytokine secretion, and proliferation.
Conjugation: Unconjugated
Isotype: IgG2a
Immunogen: Synthetic Whitlow peptide
Target background: While CAR T-cell therapies have revolutionized the treatment of B-lineage malignancies, high-resolution tracking of CAR-engineered cells within the tumor microenvironment (TME) remains a significant technical challenge, particularly in archival formalin-fixed paraffin-embedded (FFPE) tissues. To address this, murine monoclonal antibodies (mAbs) have been developed to specifically target the Whitlow linker, a synthetic peptide commonly utilized in the scFv domains of multiple FDA-approved CAR products (e.g., axi-cel, liso-cel). Because this linker is absent in native human tissue and conserved across various antigen specificities, these mAbs provide a universal tool for the in situ identification, selection, and functional analysis of CAR-expressing cells. This methodology enables a more precise evaluation of CAR T-cell infiltration, persistence, and correlation with clinical outcomes or toxicities.

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Table with Kd values of antibodies
Scientific figure containing flow cytometry histograms

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