Establishment and preliminary characterization of hybridoma cell lines secreting monoclonal antibodies against Prion Proteins.
- Author:
Li ZHAO
1
;
Rong JI
;
Jian-wei WANG
;
Chun-hui HAN
;
Xiu-ping YU
;
Xiao-ping DONG
;
Tao HUNG
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Antibodies, Monoclonal; biosynthesis; Antibodies, Viral; biosynthesis; immunology; Antibody Specificity; Cattle; Cricetinae; Cross Reactions; Encephalopathy, Bovine Spongiform; prevention & control; Enzyme-Linked Immunosorbent Assay; Female; Humans; Hybridomas; secretion; Male; Mice; Mice, Inbred BALB C; PrPSc Proteins; immunology; Prion Diseases; prevention & control; Prions; immunology; Recombinant Fusion Proteins; biosynthesis; immunology
- From: Chinese Journal of Experimental and Clinical Virology 2003;17(2):133-136
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo obtain monoclonal antibodies (McAbs) which can be widely used to detect mammalian prions (PrP) and to develop diagnostic tests for screening transmissile spongiform encephalopathies (TSE) as well as for studying pathogenesis of prion-related diseases.
METHODSBALB/c mice were immunized separately with bovine PrP peptide 29-48 (BoP1) and 89-108 (BoP2) coupled to keyhole limpt hemocyan. Two hybridoma cell lines secreting monoclonal antibodies against these peptides were established by cell fusion and 2 to 3 rounds of cell cloning. The reactions of the McAbs to the recombinant bovine (Bo)PrP(25-242), human (Hu)PrP(23-231) and hamster (Ha) PrP (23?231) were tested separately by Western blotting.
RESULTSThrough cell fusion, two hybridoma cell lines secreting McAbs against BoP1 and BoP2, designated D11 and D8 accordingly, were identified by ELISA and cell cloning. The McAbs produced by these cell lines reacted well with the recombinant PrP proteins; (Bo) PrP (25-242), (Hu) PrP (23-231), and (Ha) PrP (23-231), respectively.
CONCLUSIONSTwo McAbs reacting with bovine, human and hamster PrPs were successfully generated, they are potential to be used to detect PrPs in mammals and to study the mechanism of pathogenesis of TSE.
