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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。
OBJECTIVE: To extract and identify pig rib chondrocyte chain protein (p-CLP) and determine the affinity between p-CLP and hyaluronic acid to provide the experimental basis for the development of tumor cartilage. Methods: p-CLP was extracted from costal cartilage by affinity chromatography, and p-CLP was analyzed by SDS-PAGE gel electrophoresis and amino acid analysis. Radioimmunoassay was used to detect the expression of p-CLP and HA ) Affinity. Results: The yield of p-CLP extracted protein was 2.2%. The amino acid analysis of p-CLP showed the highest content of glutamate (2.26mg / 20mg). The radioactive in vitro binding assay showed that the binding activity of p-CLP with HA was 6.30 ± 1.73) × 109M-1. Conclusion: The high yield of p-CLP extracted by affinity chromatography and the high binding affinity between p-CLP and HA can provide experimental evidence for the prevention and treatment of tumor, ophthalmological diseases and bone diseases.