Chromatographic refolding of recombinant human interferon gamma by an immobilized sht GroEL191-345 column

Minichaperome sht GroEL191-345 was covalently coupled to NHS-activated Sepharose FastFlow gel. Refolding of recombinant human interferon gamma (rhfFN-gamma) was carried out on a chromatographic column packed with immobilized minichaperone. The effects of salt concentration, urea concentration gradient, elution flow rate and protein loading on the refolding efficiency were investigated. The results indicated that immobilized slit GroEL191-345 chromatography was an effective protocol for the refolding of rhIFN-gamma. When loading 100 mu l denatured rhIFN-gamma (17.8 mg/ml), the protein mass recovery and total activity obtained in this optimal process reached 74.25% and 6.74 x 10(6) IU/ml, respectively with the immobilized minichaperone column which was reused for 10 times with 25 % decrease of renaturation capacity. (c) 2006 Elsevier B.V. All rights reserved.