Preparation of monoclonal antibodies against prion proteins with full-length hamster PrP.
- Author:
Xin-Li XIAO
1
;
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
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Antibodies, Monoclonal; immunology; Blotting, Western; Brain; metabolism; Cell Line, Tumor; Cricetinae; Enzyme-Linked Immunosorbent Assay; Female; Immunization; Immunohistochemistry; Mice; Mice, Inbred BALB C; PrPC Proteins; genetics; immunology; PrPSc Proteins; genetics; immunology; Recombinant Proteins; immunology
- From: Biomedical and Environmental Sciences 2005;18(4):273-280
- CountryChina
- Language:English
-
Abstract:
OBJECTIVETo prepare the PrP specific monoclonal antibodies (mAbs) that can be used for the detection of mammalian prions and study of pathogenesis of prion diseases.
METHODSSeveral 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.
RESULTSThe 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 PrPSc 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. Immunohistochemistry assays confirmed that mAbs B7 and B9 could recognize not only extracellular but also intracellular PrPsSc.
CONCLUSIONThe mAbs of prion protein are successfully generated by hybridoma technique and can be applied for the diagnosis of prion associated diseases.
