嵌合蛋白sTNFRⅡ-IgG Fc纯化与生物学活性研究

Purification and Bioactivity of Chimeric Protein sTNFRⅡ-IgG Fc

目的 建立嵌合蛋白sTNFR IgGFc的纯化工艺 ,并对该蛋白的理化和生物活性进行研究。方法 嵌合蛋白sTNFRⅡ IgGFc经变性、复性后 ,用金属螯合层析进行纯化 ,纯化的蛋白进行配基结合实验和拮抗TNF活性检测。结果 该嵌合蛋白的纯度大于 95 % ,在体外能够二聚化 ,保留了sTNFRⅡ对hTNFα的结合特性 ,同单体sTNFRⅡ相比 ,对hTNFα的拮抗活性明显提高。

Objective To develop a purification process and study the physicochemical and biological activities of chimeric protein sTNFRⅡ IgG Fc.Methods After denaturation and renaturation, the chimeric protein sTNFRⅡ IgG Fc was purified by metal chelate Sepharose chromatography. The purified protein was detected for binding and neutralizing activities. Results The chimeric protein showed a purity of above 95%. It formed dimmer in vitro and remained the binding activity of sTNFRⅡ to hTNFα. Compared with that of monomer TNFRⅡ, the neutralizing activity of the chimeric protein was significantly improved.Conclusion sTNFRⅡ IgG Fc was purified, and its bioactivity was studied. This paper laid a foundation of further study on application of chimeric protein sTNFRⅡ IgG Fc.