화학공학소재연구정보센터
Biotechnology Progress, Vol.11, No.2, 208-213, 1995
Membrane-Based Cell Affinity-Chromatography to Retrieve Viable Cells
A novel scheme for the separation and live recovery of one cell type from a mixture of cells using a cell affinity chromatography (CAC) system is demonstrated. An antimurine IgG was chemically immobilized to a cellophane support via a carbonyldiimidazole (CDI) link. Murine splenocytes 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 cells. Elution of the B-cells was initiated by the transmembrane diffusion of hydrochloric acid (pH 1), supplied to the side of the membrane opposite 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 acid, were retrieved. Results showed that, on average, 250 cells/mm(2) attached to antibody immobilized on cellophane surfaces, at a shear rate of 15 s(-1), and that attached cells were successfully displaced by acid supplied to the side of the membrane opposite that holding the cells. On average, at least; 60% of the B-cells removed by this elution appeared viable, based on a Trypan Blue dye exclusion assay.