Expression of GST-HAI-1 fusion protein and development of monoclonal antibody against human hepatocyte growth factor activator inhibitor 1.
- Author:
Hai-Xia CHEN
1
;
Jiang CAO
;
Jian-Gen SHEN
;
Shu ZHENG
Author Information
1. 2nd Affiliated Hospital, Zhejiang University College of Medicine, Hangzhou 310009, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Antibodies, Monoclonal;
immunology;
Blotting, Western;
Glutathione Transferase;
genetics;
Immunohistochemistry;
Mice;
Mice, Inbred BALB C;
Proteinase Inhibitory Proteins, Secretory;
analysis;
genetics;
immunology;
Recombinant Fusion Proteins;
biosynthesis;
immunology
- From:
Chinese Journal of Biotechnology
2004;20(4):496-500
- CountryChina
- Language:Chinese
-
Abstract:
The aim of this study is to develop monoclonal antibody against human hepatocyte growth factor activator inhibitor 1 (HAI-1) for future study of HAI-1. The cDNA fragments of human hepatocyte growth factor activator inhibitor 1 (HAI-1) were subcloned to construct GST-HAI-1 fusion protein expression vectors. The vectors were transformed into E. coli and fusion protein expression was induced by IPTG. The GST-HAI-1 fusion proteins were separated on preparative SDS-PAGE and recovered by electroelution, and used to immunize BALB/c mice. Hybridomas producing monoclonal antibodies against human HAI-1 were prepared by cell fusion technique and characterized by ELISA, Western Blot and immunohistochemical staining. One hybridoma cell line, ZMC6, was obtained, which produces specific antibody against the expressed GST-HAI-1 fusion protein. The monoclonal antibody recognizes both the membrane-type and secretory-type HAI-1 proteins of colorectal tissue. The successful development of anti-HAI-1 antibody provides a powerful tool for further investigation on HAI-1's function.
