CST, 33709S, CAR (Whitlow/218 Linker) Antigen Density Quantitation Kit (PE)

Protocol
Available protocols: Flow Cytometry (Live)
Specificity / Sensitivity
Whitlow/218 Linker (E3U7Q) Rabbit Monoclonal Antibody (PE Conjugate) recognizes exogenously expressed levels of scFv-based CARs containing a Whitlow/218 linker.
Species Reactivity: All Species Expected
Background
The stoichiometric relationship between surface expressed scFv-based chimeric antigen receptors (CARs) and their cognate antigens expressed on target cells is a critical determinant of the efficacy of CAR-engineered immune cells. As such, accurate measurement of CAR density on the cell surface is of paramount importance to assess how changes to CAR structure and cell engineering protocols affect CAR expression (1-15). The Whitlow/218 peptide is a commonly used linker that connects the variable heavy (VH) domain and variable light (VL) domain of single-chain variable fragments (scFvs) and CARs that utilize an extracellular domain scFv for target antigen recognition. The Whitlow/218 linker affords the scFv with enhanced resistance to proteolysis and aggregation (16).