CHEMICAL, PHYSICAL AND CHROMATOGRAPHIC PROPERTIES OF SUPERDEX-75 PREP GRADE AND SUPERDEX-200 PREP GRADE GEL-FILTRATION MEDIA

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 beads. They consist of cross-linked agarose beads modified with dextran to obtain the desired selectivity. The mean particle size is ca. 33-mu-m (30-36-mu-m) and a reduced plate height of ca. 2.0 is regularly obtained in packed columns. A flow-rate of 50 cm/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 factor molecular weight 22 000 dalton) from dimers and oligomers. Superdex 200 prep grade was used to obtain baseline separation of a monoclonal antibody from antibody dimer and contaminating transferrin.