唾液蛋白质组学唾液样本制备的方法学初探  

An attempt to prepare saliva sample for human saliva proteome

作　　者：姜静[1] 孙其飞[2] 陈勇[3] 柳晓兰[3] 祁森荣[2] 朱红[1] 张春梅[2] 

机构地区：[1]首都医科大学附属北京儿童医院,北京100045 [2]首都医科大学口腔医学院,北京100050 [3]北京蛋白质组研究中心抗体工程研究室,北京102206

出　　处：《临床口腔医学杂志》2010年第9期515-517,共3页Journal of Clinical Stomatology

基　　金：国家自然科学基金资助(30960423)

摘　　要：目的:通过对唾液淀粉酶单克隆抗体的制备和鉴定,寻找蛋白质组学研究中样本制备关键环节的可行性方案,为日后唾液蛋白质组学研究奠定基础。方法:将腮腺液经超滤法浓缩蛋白后,进行SDS-PAGE分析,切取分子量为50～65kDa的高丰度蛋白条带,研磨后注射BALB/c小鼠,诱导产生免疫应答并制备人唾液淀粉酶单克隆抗体,应用酶联免疫吸附试验和蛋白免疫印记等方法鉴定。结果:成功制备和鉴定了人唾液淀粉酶单克隆抗体。结论:成功制备和鉴定了唾液淀粉酶单克隆抗体,为唾液蛋白质组学研究中样本制备提供解决方案。Objective：To prepare and characterize monoclonal antibodies（mAbs）against salivary amylases for providing a solution of saliva sample preparation in proteomic study of saliva.Method：Proteins in human parotid saliva were concentrated by using ultrafiltration and analyzed by SDS-PAGE.The proteins strap between 50～65 kDa was cut,grinded and used to immunize BALB/c mice.The monoclonal antibodies（mAbs）against salivary amylase in human parotid saliva were prepared and characterized by ELISA and Western blot.Result：mAbs against salivary amylases were successfully prepared and characterized.Conclusion：mAbs against salivary amylases were successfully prepared and characterized.The present study may provide a solution of saliva sample preparation for the future study of human saliva proteome.

关 键 词：腮腺液 蛋白质组学 样本制备 

分 类 号：Q812[生物学—生物工程]