肺表面活性物质蛋白B、C的提纯及生物活性功能分析

Isolation and purification pulmonary surfactant protein B,C and biological activity detection,

目的 为了提高肺表面活性物质的疗效及减少异性蛋白造成的免疫反应 ,我们对猪肺组织中的肺表面活性物质蛋白B和C进行了提取及纯化。方法 用生理盐水灌洗猪肺后 ,经低温离心 ,萃取 ,层析来提取肺表面活性物质蛋白B和C ,又用高效液相色谱纯化蛋白B和C。并用膜天平测定所提纯的肺表面活性蛋白SP -B和SP -C的生物活性。结果 用SDS -聚丙烯酰胺凝胶电泳检测 ,显示杂蛋白条带明显减少 ,分子量分别为 6 .7× 10 0 0、17.9× 10 0 0。且提纯的蛋白用较好的生物活性。P <0 .0 5。结论高效液相色谱能在分子水平上纯化肺表面活性物质蛋白B和C ,并较好地保持了其生物活性。

Objective:In order to improve the treatment effect of pulmonary surfactant and avoid the immunity reaction of protein,we isolated and purified the pig pulmonary surfactant.Methods:The pig lungs were washed with saline.The mixture were centrifugation at low temperature,reverse-phase chromatography and got protein sediment.And then we purified SP-B and SP-C by reverse-phase HPLC.Result:The samples upon SDS/polyacrylamide gel electrophoresis show multiple bands.The molecular weight is 6.7X1000 and 17.9X1000.p<0.05.Conclusion:The pulmonary surfactant of SP-B and SP-C were purified effectively by reversephase high performance liquid chromatography.