Superporous agarose as an affinity chromatography support

Superporous agarose beads were used as an affinity support in column chromatography. These beads characteristically possess two sets of pores, normal diffusion pores and flow pores, so-called superpores. The superpores, whose diameter is a substantial fraction of the particle diameter (i.e. 1/3 to 1/10 of the particle diameter), allow part of the chromatographic 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-rates [Gustavsson and Larsson, J. Chromatogr. A, 734 (1996) 231-240.] Superporous agarose beads and homogeneous agarose beads were each derivatized with two types of affinity ligands. A NAD(+) analogue was used for the purification of bovine lactate dehydrogenase and protein A was used for the adsorption of rabbit IgG. The performances of superporous beads and homogeneous beads were compared. Superporous bead columns derivatized with protein A and NAD(+) analogue could be operated 5 times and 3 times, respectively, as fast as corresponding homogeneous bead columns. (C) 1997 Elsevier Science B.V.