双向电泳中人血清蛋白样品不同处理方法的比较

山东大学学报(医学版) ›› 2009, Vol. 47 ›› Issue (11): 89-94.

双向电泳中人血清蛋白样品不同处理方法的比较

王班琴¹，秦兆宇²，柏淑美¹，刘芙军³，李芸¹，刘师莲¹

1. 山东大学医学院生物化学与分子生物学研究所，济南 250012；
2. 山东大学威海分校海洋学院，山东威海 264209；
3. 烟台市毓璜顶医院中心实验室，山东烟台 264000

收稿日期:2009-05-06 发布日期:2009-11-16
通讯作者: 刘师莲，教授，硕士生导师，主要从事神经系统疾病蛋白质组学方向研究。 Tel: 053188382346，Email:liushilian@sdu.edu.cn
作者简介:王班琴（1981- ），女，硕士研究生，主要从事蛋白质组学研究 Email:banqinwang-81@163.com

Comparison of different sample preparation methods for human serum 2DE

WANG Banqin ¹ , QIN Zhaoyu ² , BAI Shumei ¹ , LIU Fujun ³ , LI Yun ¹ , LIU Shilian ¹

1. Institute of Biochemistry and Molecular Biology, School of Medicine, Shandong University, Jinan 250012， China；
2. School of Marine, Shandong University at Weihai, Weihai 264209, Shandong, China；
3. Central Laboratory, Yantai Yuhuangding Hospital, Yantai 264000, Shandong, China

Received:2009-05-06 Published:2009-11-16

摘要/Abstract

摘要：

目的针对人血清双向电泳流程中样品制备的不同方法，本研究进行了系统的比较和分析。方法采用丙酮沉淀法，三氯醋酸沉淀法，三氯醋酸/丙酮沉淀法，蛋白纯化试剂盒，去白蛋白及免疫球蛋白试剂盒预处理血清样品；双向电泳分离；硝酸银染色；软件分析所得蛋白质斑点。结果对丙酮沉淀法，三氯醋酸沉淀法，三氯醋酸/丙酮沉淀法，蛋白纯化试剂盒处理血清所得到的2DE图谱统计分析，得到平均蛋白质点数分别为1?802±12，1?648±10，1?727±13，1?670±14；对去除白蛋白及免疫球蛋白血清所得到的双向电泳（2DE）图谱与丙酮沉淀血清所得到的图谱进行统计，得到平均蛋白质点数分别为1?212±16，1?258±12，并对其中差异蛋白进行了质谱鉴定。结论结果表明不同血清沉淀方法对血清的双向电泳各有优劣；血清去除白蛋白及免疫球蛋白之后，可以提高一些低丰度蛋白的检出，但会导致部分蛋白的丢失。

关键词: 双向电泳;样品处理;蛋白组学;血清

Abstract:

To compare different sample preparation methods for human serum 2DE. MethodsSerum samples were prepared by five different methods: acetone precipitation, trichloroacetic acid （TCA） precipitation, acetone /TCA precipitation, 2D clean up kit, and Albumin and IgG Removal Kit. Then, after the samples were run on two dimensional gel electrophoresis (2DE), 2DE gels were stained and analyzed by Image Master 5.0, and protein spots were identified by MALDITOF MS. ResultsThe total protein number of samples processed by acetone precipitation, TCA precipitation, Acetone/TCA precipitation, and 2D clean up kit was 1?802±12, 1?648±10, 1?727±13 and 1?670±14, respectively. In addition, the total protein number of samples, one of which was processed by sample preparation kits removing the albumin and immunity goblin, the other one of which was not, was 1?212±16 and 1?258±12, respectively. Conclusion2DE maps obtained from samples treated by different serum precipitation methods were different. By removing albumin and immunity goblin from the serum, low abundance proteins were increased, but some high abundance proteins were lost.

Key words: Two dimensional gel electrophoresis; Sample preparation; Proteomics; Serum

中图分类号:

Q53

王班琴1，秦兆宇2，柏淑美1，刘芙军3，李芸1，刘师莲1. 双向电泳中人血清蛋白样品不同处理方法的比较[J]. 山东大学学报(医学版), 2009, 47(11): 89-94.

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