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.
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