Application of Immobilized Cholesterol Column Chromatography in Protein Refolding

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Abstract Cholesterol modified Sepharose matrix was prepared by a two-step method with cyanuric chloride as a linker.A refolding chromatography with immobilized cholesterol was studied for the reactivation of denatured-reduced lysozyme and the optimal method for protein refolding was established.Furthermore,the effects of flow rate,initial protein concentration and loading mass on refolding efficiency were investigated.The results showed that when the loading mass of Lysozyme was 0.25 mg/mL of the support,96.3% protein recovery and 91.4% activity yield were obtained by adsorption-elution way at flow rate of 0.02 mL/min.Denatured-reduced lysozyme was effectively refolded into the active form.

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Received 2008-09-15; published 2008-09-15

Cite this article:

LI Jin,BAI Shu.Application of Immobilized Cholesterol Column Chromatography in Protein Refolding[J] Chemcial Industry and Engineering, 2008,V25(5): 404-408

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