Whitlow linker Monoclonal Antibody

The production of the Whitlow linker monoclonal antibody includes the multiple injections of the Whitlow linker-KLH into a mouse, collection of the spleen of the immunized mouse and isolation of the B cell-containing splenocytes, fusion of the B cell-containing splenocytes and immortal myeloma cells to form the hybridoma, and culture of the whitlow linker antibody-producing hybridomas. The whitlow linker monoclonal antibody is purified from culture supernatant by Protein G affinity chromatography. Its purity is greater than 95%. In a functional ELISA, immobilized scFv-based anti-CD19 CAR recombinant antibody at 2 μg/mL can bind to this anti-Whitlow linker antibody, with the EC₅₀ 0f 5.147 to 5.761 ng/mL. Apart from ELISA, this antibody has also been successfully validated in WB and FC applications.

The Whitlow linker is widely used in constructing single-chain antibody variable region gene fragments (scFv) and chimeric antigen receptors (CAR). It links the heavy chain (VH) and light chain (VL) regions to ensure the two parts are effectively combined to form a complete antibody structure with antigen-binding ability. Whitlow linker also improves the ability of scFv to resist protease degradation, making it more stable in vivo and in vitro environments. So the whitlow linker monoclonal antibody can be used in preparation, detection, functionality test (ability to activate and kill target cells), and enrichment of CAR.