Expression of GST-HAI-1 Fusion Protein and Development of Monoclonal Antibody Against Human Hepatocyte Growth Factor Activator Inhibitor 1
GST-HAI-1融合蛋白的表达及抗人HAI-1单克隆抗体的制备

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.