猪软骨链蛋白的提取及鉴定  

The extraction and identification of pig cartilage chain protein

作　　者：刘志强[1] 强永刚[1] 廖永华[1] 梁智欣[1] 

机构地区：[1]广州医学院实验核医学教研室,广东广州510182

出　　处：《现代生物医学进展》2009年第24期4618-4620,共3页Progress in Modern Biomedicine

基　　金：国家自然科学基金(30670622);广州科技计划项目(2008J1-C261);广州市教育局项目(61049)

摘　　要：目的:提取和鉴定猪肋骨软骨链蛋白(p-CLP),测定p-CLP与透明质酸的亲和力,为软骨链蛋白在肿瘤显影中提供实验依据。方法:亲和层析法从仔猪肋软骨提取软骨链蛋白(p-CLP),SDS-PAGE凝胶电泳、氨基酸分析法分析p-CLP,放射性体外结合分析测定p-CLP与透明质酸(HA)的亲和力。结果:p-CLP的抽提蛋白得率为2.2%,p-CLP氨基酸分析示谷氨酸含量最高(2.26mg/20mg),放射性体外结合实验示p-CLP与HA的结合活性KA为(6.30±1.73)×109M-1。结论:亲和层析法提取p-CLP得率高,p-CLP与HA有较高的结合力,能够为p-CLP在肿瘤、眼科疾病、骨骼疾病的防治中提供实验依据。Purpose:To extract and identificate the piglet cartilage link protein (p-CLP), to determine the affinity of p-CLP and hyaluronic acid (HA), and to provide an experimental evidence for tumor imaging. Methods:p-CLP was purified by affinity chromatograpy from piglet costal;p-CLP was identified by SDS-PAGE electrophoresis,amino acid assay;the affinity between p-CLP and HA was evaluated by in-vitro competitive binding assay. Results:The yield rate of CLP by affinity chromatography was 2.2%,amino acid assay showed that the content of Glu was highest (2.26mg/20mg), in-vitro competitive binding assay showed that the affinity constant between p-CLP and HA was(6.30±1.73)×109M-1.Conclusion:The yield rate of p-CLP by affinity chromatography was high, there was a higher binding capacity between p-CLP and HA ,the method can provide experimental evidence for the prevention and treatment of tumor,ophthalmology diasease and skeleton disease by using p-CLP.

关 键 词：猪软骨链蛋白 纯化 鉴定 

分 类 号：R341[医药卫生—基础医学]