Surface affinity chromatographic separation of blood cells. I. Separation of human and rabbit peripheral granulocytes, lymphocytes and erythrocytes using polyethylene glycol-bonded column packings.

Two different types of column packings with bonded polyethylene glycol (PEG) as the stationary phase, PEG 20M-bonded Porasil AX and bisoxirane-coupled PEG 20M-Sepharose 6B, were prepared for chromatographing human and rabbit peripheral blood cells. The best separation, especially of granulocytes and lymphocytes (separation factor 2.65), was obtained with the latter packing and a phosphate-buffered solution (pH 7.5) of 8% (w/w) dextran as the mobile phase. The selectivity of the column for blood cells depended on the concentration and molecular weight of dextran and on the ionic composition of the electrolytes in the mobile phase. The recoveries of human granulocytes, lymphocytes and erythrocytes by this chromatographic system were about 67, 82 and 50%, respectively.