摘要
用层析和制备SDS-PAGE法纯化舟山眼镜蛇(Najanajaatra Cantor)毒神经生长因子(NGF),免疫家兔获得抗血清。用辛酸-硫酸铵沉淀法初步纯化IgG,蛋白A-Sepharose亲和层析进一步纯化IgG,并与CNBr活化的Sepharose4B偶联,采用亲和层析法对舟山眼镜蛇毒神经生长因子进行分离纯化。产物经过SDS-PAGE检测呈一条带,并显示了良好的生物学活性。纯化NGF最大比活性为5.0×104U/mg蛋白,亲和常数为4.35×108L/mol,亲和层析分离NGF得率比传统分离方法得率提高35.2%,该方法为NGF的大量提取提供了技术支持。
Nerve growth factor（NGF） from Naja naja atra Cantor venom was purified by chromatography and SDS - PAGE. Antiserum against NGF was prepared by immunized rabbits. IgG in the antiserum was purified by octanoic acid -ammonium sulfate precipitation and Protein A -Sepharose chromatography. Purified IgG was coupled to CNBr activated Sepharose 4B and subsequently used to purify NGF from crude Naja naja atra Cantor venom. The SDS - PAGE analysis of the purified NGF preparation showed one band by protein staining. It showed significantly biological activity and the specific activity of NGF was 5.0 ×10^4U/mg protein. The affinity constant of IgG and NGF determined by ELISA was 4. 35×10^8L/mol. NGF weight obtained by immunoaffinity chromatography was 35.2% higher than that by traditional purification method. This method was technical support for purifying NGF in great scale.
出处
《生物学杂志》
CAS
CSCD
2007年第4期41-44,共4页
Journal of Biology
基金
安徽省教育厅自然科学基金项目资助(2006KJ415B)
关键词
舟山眼镜蛇
神经生长因子
亲和层析
：Naja naja atra Cantor
nerve growth factor
immunoaffinity chromatography
