Chemical, physical and chromatographic properties of Superdex 75 prep grade and Superdex 200 prep grade gel filtration media

Abstract Two novel gel filtration media, Superdex 75 prep grade and Superdex 200 prep grade are described. The new media have high selectivities similar to those of Sephadex G-75 and G-200, respectively, but permit the use of high flow-rates as with cross-linked agarose hbeads. They consist of cross-linked agarose beads modified with dextran to obtain the desired selectivity. The mean particle size is ca. 33 μm (30–36 μm) and a reduced plate height of ca. 2.0 is regularly obtained in packed columns. A flow-rate of 50cm/h can be used even in large columns. The media are chemically stable at pH 3–14 and can be used with a wide range of eluents. The performance of the media with respect to flow-rate, sample load and sample concentration for given model systems is described. Superdex 75 prep grade was used to separate monomeric forms of a recombinant fusion protein (ZZ—brain insulin-like growth molecular weight 22 000 dalton) from dimers and oligomers. Superdex 200 prep grade was used to obtain separation of a monoclonal antibody from antibody dimer and contaminating transferrin.