Separation of B- and T-lymphocytes by cellular adsorption chromatography using poly(2-hydroxyethyl methacrylate)/polyamine graft copolymer as column adsorbent

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Abstract
A series of poly(2-hydroxyethyl methacrylate)/polyamine graft copolymers (HA copolymers) were examined as an adsorbent for cellular adsorption chromatography in order to separate cell populations with high efficacy. The capability of each copolymer sample to selectively adsorb rat lymphocyte subpopulations (B-cells and T-cells) was evaluated by applying a lymphocyte suspension to a copolymer-coated glass bead column. A separation factor (AB/AT) greater than 5.0 was achieved for the HA copolymer, compared to 1.9 and 1.1 for homopolymers of poly(2-hydroxyethyl methacrylate) and of a polyamine (poly(N,N-diethylaminoethylstyrene)), respectively. By loading a rat lymphocyte suspension containing B-cells (28%) and T-cells (72%) through the HA copolymer column, more than 60% of T-cells in 90% purity or more were recovered in the effluent.