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唾液蛋白质组学唾液样本制备的方法学初探
引用本文:姜静,孙其飞,陈勇,柳晓兰,祁森荣,朱红,张春梅.唾液蛋白质组学唾液样本制备的方法学初探[J].临床口腔医学杂志,2010,26(9):515-517.
作者姓名:姜静  孙其飞  陈勇  柳晓兰  祁森荣  朱红  张春梅
作者单位: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
基金项目:国家自然科学基金资助项目 
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。

关 键 词:腮腺液  蛋白质组学  样本制备

An attempt to prepare saliva sample for human saliva proteome
JIANG Jing,SUN Qi-fei,CHEN Yong,LIU Xiao-lan,QI Sen-rong,ZHU Hong,ZHANG Chun-mei.An attempt to prepare saliva sample for human saliva proteome[J].Journal of Clinical Stomatology,2010,26(9):515-517.
Authors:JIANG Jing  SUN Qi-fei  CHEN Yong  LIU Xiao-lan  QI Sen-rong  ZHU Hong  ZHANG Chun-mei
Institution:JIANG Jing1,SUN Qi-fei2,CHEN Yong3,LIU Xiao-lan3,QI Sen-rong2,ZHU Hong1,ZHANG Chun-mei2.1.Capital Medical University,Beijing Children's Hospital.Beijing 100045,China,2.School of Stomatology,Capital Medical University,Beijing 100050,3.Department of Antibody Engineering,Beijing Proteome Research Center,Beijing 102206,China.
Abstract: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.
Keywords:parotid saliva  proteom  sample preparation  
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