SELDI技术对肝硬化相关血浆差异蛋白的鉴定  

Identification of liver cirrhosis related biomarkers founded by SELDI technology

作　　者：陆晔[1,2,3] 刘杰[4] 项翠琴[3] 林成招[2] 王吉耀[4] 

机构地区：[1]复旦大学化学系,上海200433 [2]复旦大学生物医学研究院系统生物学研究室 [3]上海市疾病预防控制中心公共卫生分子生物学研究室 [4]复旦大学附属中山医院消化内科

出　　处：《中国实验诊断学》2009年第10期1311-1314,共4页Chinese Journal of Laboratory Diagnosis

基　　金：上海市创新团队项目(03DZ14024)

摘　　要：目的建立基于SELDI-TOF-MS技术的蛋白质分离、纯化、鉴定技术,用于鉴定运用SELDI技术发现的肝硬化相关血浆差异蛋白。方法采用尿素预处理血液样本,结合离子交换层析、SELDI芯片检测及芯片上酶解、质谱鉴定等多种技术手段,对肝硬化相关的差异蛋白质进行了分离、纯化和鉴定。结果选用阳离子交换剂对血浆样品进行分离,并对分离的各组分进行芯片检测,在300 mM NaCl洗脱组分中,含有与目标蛋白荷质比相同、理化性质相同的蛋白质峰,并且该目标蛋白得到富集。采取芯片原位酶解的方法对目标蛋白进行快速酶解和质谱鉴定,鉴定该差异蛋白为人血浆结合珠蛋白。结论这一技术路线可以对前期运用SELDI技术检测到的疾病相关的差异蛋白质峰进行分离、纯化和鉴定,从而开展进一步的机制研究和应用研究。Objective To identify the liver cirrhosis related biomarkers found by SELDI-TOF-MS technology.Methods Various technologies,including ion exchange,SELDI chip confirmation,on chip digestion and mass spectrum identification,were used to isolate,purificate and identify the liver cirrhosis related biomarkers.Results The target protein was enriched by cation exchange in the fraction of 300 mM NaCl.After on chip digestion and mass spectrum identification,the protein was identified as human Haptoglobin.Conclusion The technology described here can be used to identify the disease related biomarkers found by SELDI technology,which will facility the further mechanism and application research.

关 键 词：SELDI技术 血浆蛋白质组 疾病标志物 蛋白质鉴定 

分 类 号：R575.2[医药卫生—消化系统]