Cell Affinity Chromatography

Cell affinity separations using magnetically stabilized fluidized beds: Erythrocyte subpopulation fractionation utilizing a lectin‐magnetite support

“…The low recovery of bound A erythrocytes using this method was probably caused by a high surface concentration of HpA. The elution concentration of D-GalNAc in this study (30 mg/mL) is 15 times higher than what we found necessary to recover >90% of bound A erythrocytes from a commercial support, HpA-Sepharose 6MB, available from Pharmacia Biotech (Putnam et al, 1999). In addition to a higher amount of HpA used in the coupling step for magnetite (10 mg per 3 mL of sedimented support versus 3.6 mg per mL of sedimented Sepharose 6MB), the nonporous nature of magnetite compared to porous Sepharose likely led to a significantly higher HpA surface concentration.…”

“…Additionally, the HpA support facilitated the processing of approximately 1.1 × 10 9 cells in 2 h. The support was used four times without significant loss of activity. The processing rate of even this unoptimized system is comparable to packed bed CAC systems (Putnam et al, 1999).…”

“…Due to their low cost and simple operation, cell affinity chromatography (CAC) systems are increasingly being used for the isolation of a variety of immune cell types (Putnam et al, 1999). CAC relies on the interaction of cell surfacebound molecules and their complementary ligands (monoclonal antibodies, lectins).…”

“…In addition to selective agglutination, lectins have also found extensive use in packed bed cell affinity chromatography systems (Putnam et al, 1999;Sharma and Mahendroo, 1980). The advantage of packed bed CAC over agglutination is the increase in throughput of the system.…”