SUPERPOROUS AGAROSE AS AN AFFINITY-CHROMATOGRAPHY SUPPORT

Superporous agarose beads were used as an affinity support in column c hromatography. These beads characteristically possess two sets of pore s, normal diffusion pores and flow pores, so-called superpores. The su perpores, whose diameter is a substantial fraction of the particle dia meter (i.e. 1/3 to 1/10 of the particle diameter), allow part of the c hromatographic flow to pass through each individual bead. Consequently , significant improvement in mass transfer is observed in superporous beads as compared with homogeneous beads, especially at high flow-rate s [Gustavsson and Larsson, J. Chromatogr. A, 734 (1996) 231-240.] Supe rporous agarose beads and homogeneous agarose beads were each derivati zed with two types of affinity ligands. A NAD(+) analogue was used for the purification of bovine lactate dehydrogenase and protein A was us ed for the adsorption of rabbit IgG. The performances of superporous b eads and homogeneous beads were compared. Superporous bead columns der ivatized with protein A and NAD(+) analogue could be operated 5 times and 3 times, respectively, as fast as corresponding homogeneous bead c olumns. (C) 1997 Elsevier Science B.V.