Preparation of a monoclonal antibody against fusion protein preM/EIII of WNV.
- Author:
Linhai LI
1
;
Lidan CHEN
;
Yang LIAO
;
Jianyun CHEN
;
Yuling SHI
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Antibodies, Monoclonal; immunology; Blotting, Western; Cross Reactions; Hybridomas; immunology; Mice; Mice, Inbred BALB C; Viral Fusion Proteins; immunology; West Nile virus; immunology
- From: Journal of Southern Medical University 2012;32(5):742-745
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo prepare a monoclonal antibody (mAb) against the fusion protein preM/EIII of West Nile virus (WNV) for clinical detection of WNV.
METHODSSp2/0 cells were fused with the spleen cells of BALB/c mice immunized with the recombinant fusion protein preM/EIII expressed in E. coil to obtain the hybridoma cell line that secreted preM/EIII mAb. The hybridoma cells were injected into the peritoneal cavity of BALB/c mice and the ascites was collected and purified. The specificity and titer of the obtained mAb were determined using ELISA and Western blotting.
RESULTSOne hybridoma cell line secreting preM/EIII mAb, named ME1, was obtained. The titer of the purified mAb was 10(-6). Identified as a mAb of the Ig subclass IgG1, ME1 was capable of specific reactions with preM/EIII protein and WNV without cross-reactions with other viruses such as JEV, SLEV, YFV and DENV. The accuracy of clinical testing of MNV with ME1 was 97.78%.
CONCLUSIONThe mAb against preM/EIII obtained have a high specificity and accuracy in clinical testing of MNV and can be used in clinical diagnosis of MNV infection.
