人胰液双向凝胶电泳分离技术建立及优化.docVIP

人胰液双向凝胶电泳分离技术建立及优化摘要：目的建立并优化人胰液蛋白质组的双向凝胶电泳分离方法。方法通过比较几种常用的体液处理方法，确定了一种适于人胰液的蛋白质样品处理方法。经过一维等电聚焦和二维SDS 电泳后，有效分离了人胰液蛋白质组。 结果通过优化胰液蛋白质提取、上样量和固相IPG胶条分离范围的选择等步骤，建立了人胰液双向凝胶电泳的方法，并获得了分辨率较高、重复性较好的双向凝胶电泳图谱。 结论丙酮沉淀与双向凝胶电泳技术的结合是研究人胰液蛋白质组的一种有效的方法。胰液双向凝胶电泳方法的优化可为今后开展疾病相关的胰液蛋白质组学研究提供技术基础。 关键词：蛋白质组学；人胰液；双向凝胶电泳 中图分类号：O655.4文献标识码：A文章编号：1672-979X(2007)04-0005-05 Establishment and Optimization of Separation of Human Pancreatic Juice by Two-Dimensional Gel Electrophoresis LIU Xiao-yong, HAN Jin-xiang, CUI Ya-zhou, ZONG Mei-juan, CHEN Yu, TIAN Mei （Shandong Medical Biotechnological Center, Key Laboratory for Biotech-Drugs, Ministry of Health, Jinan 250062, China） Abstract：Objective To establish and optimize the two-dimensional gel electrophoresis separation of proteome in human pancreatic juice.Methods By comparing several commonly used protein extraction methods，a method for the separation of proteome in human pancreatic juice was determined. The proteome in human pancreatic juice was effectively concentrated and separated by isoelectric focusing on first dimension and SDSon second dimension.Results By optimizing the extraction of proteins in human pancreatic juice, loading amount and separation scope of IPG strip, the proteins were successfully extracted from human pancreatic juice and were separated by two-dimensional gel electrophoresis with higher resolution and better reproducibility.ConclusionAcetone-precipitation combined with two-dimensional gel electrophoresis is a suitable approach to study the proteome in human pancreatic juice. The optimization of the separation of human pancreatic juice by two-dimensional gel electrophoresis lays a technological foundation for the further investigation of disease-related human pancreatic juice proteomics. Key words：proteomics; human pancreatic juice; two-dimensional gel electrophoresis 胰液中含有胰腺组织细胞所释放的高浓度的蛋白质与DNA。胰液中蛋白质的变化与胰腺的生理或病理状态密切相关，这些蛋白质是潜在的胰腺疾病标记物。因此，胰液的检测对早期胰腺疾病的筛查具有重要意义。目前国际上对胰液蛋白组分的研究多采用表面增强激光解析飞行时间质谱（SELDI-