[Expression of GST-HAI-1 fusion protein and development of monoclonal antibody against human hepatocyte growth factor activator inhibitor 1].
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.
Chen
Hai-Xia
HX
2nd Affiliated Hospital, Zhejiang University College of Medicine, Hangzhou 310009, China.
Cao
Jiang
J
Shen
Jian-Gen
JG
Zheng
Shu
S
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D015153:Blotting, Western; D005982:Glutathione Transferase / Q000235:genetics; D007150:Immunohistochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D053491:Proteinase Inhibitory Proteins, Secretory / Q000032:analysis / Q000235:genetics* / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000276:immunology
D000911:Antibodies, Monoclonal; D053491:Proteinase Inhibitory Proteins, Secretory; D011993:Recombinant Fusion Proteins; C109149:SPINT1 protein, human; D005982:Glutathione Transferase