Purified Monoclonal Antibody of Anti-Flavivirus Group Antigen Clone D1-4G2-4-15 for the Development of Japanese Encephalitis Virus Surface Proteins in Plants

Preeyanuch Sayboonruan, Wipa Tangkananond, Wassana Wijagkanalan, Butsaya Thaisomboonsuk


Production of Japanese encephalitis virus (JEV) surface proteins in plants requires monoclonal antibodies specific to some part of antigens of JEV as a tool of detection. This research is to develop anti-flavivirus group antigen clone D1-4G2-4-15 monoclonal antibodies specific to the envelope protein of viral particle of genus Flavivirus in serum-free media by inside adaptation culture method. The monoclones grew quickly in serum-free media and were purified from cell culture supernatant by precipitation with 50% saturated ammonium sulfate and fractionated by affinity chromatography. Then, the product was tested for antibody specificity by typing ELISA. The test results were positive with inactivated Japanese encephalitis vaccines and Dengue virus type 1. The stability tests were performed at -20°C for 10 weeks.  The detection of antibodies against the Dengue virus type 1 can be used at a dilution of 1: 1000. The cost of production is cheaper than the cost of antibodies production by culturing cells in serum media. This technique is feasible for the laboratory because of its simplicity. The monoclonal antibodies derived from this research are stable and specific to Flavivirus antigens. It can be used to study the properties of the JEV surface protein production in plants and other research of the genus Flavivirus.


Keywords: Japanese encephalitis virus, D1-4G2-4-15, serum-free media, purification.

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