Generation of Monoclonal Antibodies Against Human Papillomavirus Type16 E7 Protein : Usefulness for Various E7 Detection Systems.
- Author:
Woo Hyun CHANG
;
Min Kee CHO
;
Sun Ho KEE
;
Yoon Won KIM
;
Jae Chul LEE
;
Kun Hong KIM
;
Byung Gie KIM
;
Sang Yoon PARK
- Publication Type:Original Article
- MeSH:
Animals;
Antibodies, Monoclonal*;
Bacteriophage T7;
Blotting, Western;
Clone Cells;
COS Cells;
Cytoplasm;
Escherichia coli;
Fluorescent Antibody Technique;
Humans*;
Immunization;
Maltose-Binding Proteins;
Mice;
Microscopy, Electron
- From:Journal of the Korean Society for Microbiology
1997;32(3):335-342
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
The gene encoding E7 oncoprotein of human papillomavirus type 16 was cloned and expressed in Escherichia coli, and the monoclonal antibodies (Mabs) against this expressed protein were generated. For the efficient immunization, two kind of recombinant E7 protein in fusion form were produced. One was maltose binding protein (MBP) fusion type (MBP-E7) and the other was T7 phage gene 10 product fusioa type (gene 10-E7). Immunization with these two fusion protein to mice, finally two Mabs (VD6 and IB10) were obtained. VD6 and IB10 showed reactivities with E7 protein in CaSki cell but not in HeLa by Western blot analysis. In addition, the Mab, VD6, reacted with COS-7 cell transfected with E7 gene majorly in cytoplasm by immunofluorescence test. Also VD6 could detect E7 protein in cytoplasm and nucleus of CaSki ceU by immunogold electron microscopy. Based on these results, the Mab VD6 was could be used for various E7 detection system such as Western blot analysis and immunohistochemical methods.
