GST-HAI-1融合蛋白的表达及抗人HAI-1单克隆抗体的制备

制备抗人肝细胞生长因子激活物抑制因子（HAI-1）单克隆抗体，为对HAI-1进行进一步的研究打下基础。将人HAI-1 cDNA分段克隆，构建GST-HAI-1融合蛋白原核表达载体，转化大肠杆菌后加IPTG诱导融合蛋白表达，经制备型SDS-PAGE法分离表达的GST-HAI-1融合蛋白，通过割胶、电洗脱回收融合蛋白，并以此为抗原免疫BALB/c小鼠，应用细胞融合技术制备产生抗人HAI-1单克隆抗体的杂交瘤，以ELISA、Western blot和免疫组织化学染色进行鉴定。最终获得抗人HAI-1单克隆抗体杂交瘤细胞株ZMC6，产生的单克隆抗体可特异性地与表达的GST-HAI-1融合蛋白反应，并可识别大肠组织中的膜型及脱落型HAI-1蛋白。该单克隆抗体的制备成功，为深入研究HAI-1的功能提供了有力工具。

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.