MEMBRANE-BASED CELL AFFINITY-CHROMATOGRAPHY TO RETRIEVE VIABLE CELLS

A novel scheme for the separation and live recovery of one cell type f rom a mixture of cells using a cell affinity chromatography (CAC) syst em is demonstrated. An antimurine IgG was chemically immobilized to a cellophane support via a carbonyldiimidazole (CDI) link. Murine spleno cytes flowed over the support, and B-cells were allowed to attach at a shear rate of 15 s(-1). Once loading was terminated, the support was washed at a shear rate of 315 s(-1) to remove nonspecifically bound ce lls. Elution of the B-cells was initiated by the transmembrane diffusi on of hydrochloric acid (pH 1), supplied to the side of the membrane o pposite the cells. At the same time, a shear flow of normal saline was established on the cell side of the membrane, and cells, freed by aci d, were retrieved. Results showed that, on average, 250 cells/mm(2) at tached to antibody immobilized on cellophane surfaces, at a shear rate of 15 s(-1), and that attached cells were successfully displaced by a cid supplied to the side of the membrane opposite that holding the cel ls. On average, at least; 60% of the B-cells removed by this elution a ppeared viable, based on a Trypan Blue dye exclusion assay.