Preparation of Monoclonal Antibodies Against Prion Proteins With Full-length Hamster PrP  

作　　者：XIN-LI XIAO HUI-YING JIANG JIN ZHANG JUN HAN KAI NIE XIAO-BO ZHOU YIN-XIA HUANG LAN CHEN WEI ZHOU BAO-YUN ZHANG YONG LIU XIAO-PING DONG 

机构地区：[1]School of Medicine, Xi＇an Jiao-Tong University, Zhu-Que Street 205, Xi＇an 710061, Shaanxi, China [2]National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Hngxin Rd. 100, Beijing 100052, China [3]National Laboratory of Medical Molecular Biology, Institute of Basic Medical Science, Chinese Academy of Medical Sciences and Peking Union Medical College, Dong Dan San Tiao 5, Beijing 100005, China

出　　处：《Biomedical and Environmental Sciences》2005年第4期273-280,共8页生物医学与环境科学（英文版）

基　　金：ThisworkwassupportedbytheNationalNaturalScienceFoundationofChina30070038and30130070,NationalHigh-techDevelopmentProject(863Project)2001AA215391,andEUProjectQLRT200001441.

摘　　要：To prepare the PrP specific monoclonal antibodies （mAbs） that can be used for the detection of mammalian prions and study of pathogenesis of prion diseases. Methods Several BALB/c mice were immunized with recombinant hamster prion protein （HaPrP）. Three hybridoma cell lines designated as B7, B9, and B10, secreting monoclonal antibodies against HaPrP, were established by hybridoma technique. The mAbs reactivities were evaluated with ELISA, Western blot, and immunohistochemistry. Results The mAbs produced by these cell lines reacted well with different recombinant hamster PrP proteins. Western blot analyses showed that mAbs B7 and B9 reacted with PrP^Sc from the scrapie-infected animals after proteinase K digestion with three glycosylated forms. The mAbs exhibited cross-reactivity with various PrPc from several other mammalian species, including humans and cattles, lmmunohistochemistry assays confirmed that mAbs B7 and B9 could recognize not only extracellular but also intracellular PrPso. Conclusion The mAbs of prion protein are successfully generated by hybridoma technique and can be applied for the diagnosis of prion associated diseases.To prepare the PrP specific monoclonal antibodies （mAbs） that can be used for the detection of mammalian prions and study of pathogenesis of prion diseases. Methods Several BALB/c mice were immunized with recombinant hamster prion protein （HaPrP）. Three hybridoma cell lines designated as B7, B9, and B10, secreting monoclonal antibodies against HaPrP, were established by hybridoma technique. The mAbs reactivities were evaluated with ELISA, Western blot, and immunohistochemistry. Results The mAbs produced by these cell lines reacted well with different recombinant hamster PrP proteins. Western blot analyses showed that mAbs B7 and B9 reacted with PrP^Sc from the scrapie-infected animals after proteinase K digestion with three glycosylated forms. The mAbs exhibited cross-reactivity with various PrPc from several other mammalian species, including humans and cattles, lmmunohistochemistry assays confirmed that mAbs B7 and B9 could recognize not only extracellular but also intracellular PrPso. Conclusion The mAbs of prion protein are successfully generated by hybridoma technique and can be applied for the diagnosis of prion associated diseases.

关 键 词：PRIONS Hamster prion protein Monoclonal antibodies 

分 类 号：Q959.837[生物学—动物学]