Surface affinity chromatographic separation of blood cells. II. Influence of mobile phase composition on the chromatographic behaviour of human peripheral blood cells on polyethylene glycol-bonded sepharose.

The influence of mobile phase composition on the chromatographic behaviour of human platelets, granulocytes, lymphocytes and erythrocytes has been studied by using a bisoxirane-coupled polyethylene glycol 20M-Sepharose 6B column at pH 7.5. Lowering of the concentration of dextran T40 from 8 to 2% (w/w) produced the highest separation factor between platelets and granulocytes. Addition of 0.5% (w/w) of DEAE-dextran to a mobile phase containing 2% dextran T40 or T500 increased the retention of platelets, and discriminated the cells from erythrocytes. Addition of sodium chloride increased the retention volumes of lymphocytes, granulocytes and platelets. These blood cells were adsorbed to the column in isotonic phosphate-buffered eluent, whereas in imidazole-buffered eluent about 0.15-0.154 M sodium chloride improved their resolution.