PREPARATION AND IDENTIFICATION OF MONOCLONAL ANTIBODIES AGAINST THE EXTRACELLULAR DOMAIN OF INTEGRIN

来源 :Chinese Journal of Cancer Research | 被引量 : 0次 | 上传用户:mengyan902
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Objective: To prepare monoclonal antibody (McAb) against the Integrin a6 extracellular domain and identify its biological activities. Methods: Fusion-protein of integrin a6 extracellular domain (GST-IAGED) was expressed in E.coli. JM109 and used for immunizing BALB/C mice. The spleen cells from immunized mice were fused with SP2/0 cells and selectively cultured with HAT medium. ELISA and immunocytochemistry staining were used to select hybridomas. Results: One strain of hybridoma cells that secreted specific monoclonal antibody against integrin a6 extracellular domain was indentified. The immunoglobulin subclass of the McAb was IgG1. Conclusion: The McAb against the extracellular domain of integrin a6 was successfully prepared by using GST-IA6ED fusion protein expressed by E.Coli. And the McAb had positive reaction with human hepatocarcinoma cells-BEL-7402. Methods: Fusion-protein of integrin a6 extracellular domain (GST-IAGED) was expressed in E. coli. JM109 and used for immunizing BALB / C mice. The spleen cells from immunized mice were fused with SP2 / 0 cells and optionally cultured with HAT medium. ELISA: immunofluorescent staining of the hybridomas was indentified. The immunoglobulin subclass of the McAb was IgG1. Conclusion: The McAb against the extracellular domain of integrin a6 was successfully prepared by using GST-IA6ED fusion protein expressed by E. coli. And the McAb had positive reaction with human hepatocarcinoma cells- BEL-7402.
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