Establishment and preliminary characterization of hybridoma cell lines secreting monoclonal antibodies against Prion Proteins
Zhao, L.; Ji, R.; Wang, J-wei.; Han, C-hui.; Yu, X-ping.; Dong, X-ping.; Hung, T.
Zhonghua Shi Yan He Lin Chuang Bing du Xue Za Zhi 17(2): 133-136
2003
ISSN/ISBN: 1003-9279 PMID: 12869993 Document Number: 559738
An investigation was conducted to obtain monoclonal antibodies which can be widely used to detect mammalian prion (PrP) and to develop diagnostic tests for screening transmissible spongiform encephalopathy (TSE) as well as for studying the pathogenesis of prion-related diseases. BALB/c mice were immunized separately with bovine PrP peptide 29-48 (BoP1) and 89-108 (BoP2). Two hybridoma cell lines secreting monoclonal antibodies against these peptides were established by cell fusion and 2-3 rounds of cell cloning. The reactions of monoclonal antibodies to recombinant bovine PrP(25-242), human PrP(23-231), and hamster PrP(23-231) were tested separately by Western blotting. Through cell fusion, two hybridoma cell lines secreting monoclonal antibodies against BoP1 and BoP2, designated D11 and D8, respectively, were identified by ELISA and cell cloning. The monoclonal antibodies produced by these cell lines reacted well with the recombinant PrP proteins. It is concluded that the two monoclonal antibodies reacting with bovine, human and hamster PrPs are successfully generated and these are potentially useful in detecting PrPs in mammals and in studying the mechanism of TSE pathogenesis.