Production and Characterization of Monoclonal Antibodies Against Gp Protein of Ebola Virus

The DNA fragment encoding predicted main antigenic region, aa 1-300 on Gp protein of Ebola virus (EBOV) was cloned into the vector pGEX-KG. The recombinant GST-tagged Gp-300 was expressed in Escherichia coli BL21 (DE3) by induction with 1 mM isopropyl-1-thio-b-d-galactoside and purified by dialysis. Four monoclonal antibodies (mAbs) named 1C4, 2A3, 2G7, and 2H9 against Gp protein were generated by fusing mouse myeloma cell line SP2/0 with spleen lymphocytes from Gp-300 protein-immunized mice. The activity of the mAbs was then characterized by enzyme-linked immunosorbent assay, indirect immunofluorescent assays (IFA), and western blot analysis. The results demonstrated that all the mAbs showed high specificity and sensitivity in IFA and in western blot analysis, which indicated that these mAbs against Gp protein of EBOV may be used as valuable tools for analysis of the protein functions and pathogenesis of EBOV.