Use of size-exclusion chromatography for refolding recombinant proteins from inclusion bodies

Refolding recombinant proteins from inclusion bodies is subject to low yields resulting from aggregation of folding intermediates. Redn. in aggregation may be achieved through refolding using a gel filtration matrix. The SEPROS (Size Exclusion Protein Refolding System) technique allows buffer exchange, refolding and aggregate removal in a single operation. Refolding high concns. of model proteins, such as lysozyme and carbonic anhydrase at high concns. (>20 mg/mL) has been successful (Batas and Chaudhuri, Biotech Bioeng 50, 16-23, 1996). The use of this technique to refold urokinase plasminogen activator (uPA) from inclusion bodies will be reported, along with a description of the effect of protein concn., gel filtration matrix type and sample loading on the refolding yield. A quant. mechanism for protein refolding using size exclusion chromatog. will also be presented. [on SciFinder (R)]