胶原蛋白的鉴别及其杂蛋白检测技术

目的: 建立用SDS-聚丙烯酰胺凝胶电泳法(SDS-PAGE)来鉴别胶原蛋白及分析杂蛋白含量的方法。 方法: 利用特异性的胶原酶对胶原蛋白进行酶解,而后检测样品中没有被胶原酶消解的杂蛋白的含量,同时检测实验方法的染色灵敏度,确定杂蛋白的检测下限。 结果: 经过多次验证实验表明,在本实验条件下,考马斯亮蓝对BSA的染色极限能达到100 ng,含量在0.5%以上的杂蛋白均能被测定。 结论: 本研究建立了有效的杂蛋白检测技术,为胶原蛋白为基础的生物材料中杂蛋白的检测和监控提供了有效可行的方法。

Objective :In this paper collagen was identified,and impurity protein contained in collagen was also determined by SDS-PAGE. Methods: The collagen sample's zones were compared to those of standard collagen. Results: By using collagenase digestion which specifically hydrolyze collagen,we detected the contents of impurity protein which was not been hydrolyzed by collagenase and the dyeing sensitivity,and the lower limit was been decided.These results showed that the dyeing limit of comassie blue was 100 ng,and the impurity protein with more than 0.5% content could be detected. Conclusion: In this study,we established examination method of impurity protein in collagen-based biomaterials.