CAR (G4S Linker) Cell Enrichment Kit

All components in this kit are stable in accordance with the date printed on the outer packaging label when stored at the recommended temperature.
Releasable Streptavidin Beads are supplied as 4 × 10⁸ beads/mL in phosphate buffered saline (PBS), pH 7.4, containing 0.1% Tween and 0.02% sodium azide as a preservative.
Biotinylated G4S Linker (E702V) Rabbit mAb is supplied in PBS (pH 7.2), less than 0.1% sodium azide, and 2 mg/mL BSA. Store at 4°C. Do not aliquot the antibody. Do not freeze.

Biotinylated G4S Linker (E7O2V) Rabbit mAb recognizes exogenously expressed levels of scFv-based CARs containing a G4S linker.

The CAR (G4S Linker) Cell Enrichment Kit is designed to positively select cells that are engineered to express a G4S linker-containing single-chain variable fragment (scFv)-based chimeric antigen receptor (CAR) on the cell surface. Using an indirect magnetic bead-based immunoaffinity enrichment protocol, this kit yields highly pure and viable CAR positive cells that are bead-free. Within a heterogeneous population of cells, desired CAR expressing cells are labeled with a biotinylated rabbit monoclonal antibody targeting the G4S linker. Releasable Streptavidin Beads are then incubated with the cell mixture. Bead-bound cells are isolated without columns using a magnet (not included), and then beads are removed from cells using DNase I. Following release from beads, the positively selected fraction of CAR positive cells containing surface-bound biotinylated antibody-streptavidin complexes are available for expansion or use in downstream applications such as flow cytometry and gene expression analyses.

Bead Specifications:
Releasable Streptavidin Beads are paramagnetic polystyrene beads (4.5 μm diameter) coupled to streptavidin via a DNA linker susceptible to cleavage by DNase I. The beads provided in this kit are not recommended for the isolation of phagocytic cells as the beads are likely to be engulfed by these cells.