不同样品制备方法对血浆双向凝胶电泳图谱的影响

Sample preparations for two dimensional electrophoresis map of plasma

目的探讨不同的样品制备方法对多发性大动脉炎患者血浆蛋白质组的双向凝胶电泳图谱的影响，为鉴定差异表达蛋白提供优化条件。方法采用加入SDS加热法和离心超滤方法制备方法对血浆进行处理，和未经任何处理直接上样，以90μg等上样量进行电泳，电泳方法和常规的100μg银染和1000μg考马斯亮蓝染色方法的双向凝胶电泳的图谱的图像、蛋白点数等进行对比。结果SDS加热法和离心超滤能明显改善图谱中白蛋白高丰度蛋白周围的显示。SDS加热法和离心超滤法90μg银染显示的蛋白点数分别为892．17±162．99，869．50±95．62，695．67±84．11。结论对于多发性大动脉炎血浆采用SDS加热和离心超滤进行血浆样品制备，能明显改善血浆双向凝胶电泳图谱，以90μg血浆上样量的银染方法较为合适。

Objective To discuss the impact of sample preparations on 2D electrophoresis plasma map of patients with takayasus arteritis to optimize expression of plasma proteomc for differentia analysis. Methods Sample preparations of SDS with concomitant heating and centrifugal ultrafiltration, and different loading amounts of plasma proteins were performed in 2D dimensional electrophoresis. Two dimensional electrophoresis maps were compared with those of untreated sample Results Sample preparations of SDS with concomitant heating and centrifugal ultrafiltration could improve the map resolution presenting more protein spots around high abundance protein such as albumin. Number of protein spots in map of SDS with concomitant heating and centrifugal ultrafiltration with mean of （892.17 ± 162.99 ）, （869.50 ± 95.62） respectively, was more than that of untreated samples with mean of （695.67 ± 84.11 ）. Conclusion Sample preparations with concomitant heating and centrifugal ultrafiltration prior to electrophoresis optimize the electrophoresis imaging, and loading amount of 90 μg with silver staining is appropriate for 2D electrophoresis for plasma of patients with takayasus arteritis.