THE PRODUCTION AND PURIFICATION OF CANDIDATE RECOMBINANT CHIMERIC IgA-IgG AGAINST DENGUE VIRUS IN CHO-K1 CELLS

DWI PURNO WIDEKDO, Prof. dr. Sofia Mubarika, M.Med.Sc.

2014 | Tesis | S2 Bioteknologi

In recent years, the production of recombinant pharmaceutical proteins in mammalian systems has increased significantly such as antibody. Antibodies are the important molecules of the host protective response against virus infection with the potentially contribute to immunity via several distinct mechanisms, including an ability to directly neutralize virus infection. Aim of this research is to establish recombinant Chinese Hamster Ovary (CHO) to produce chimeric IgA-IgG. IgA and IgG is the most responsible antibody class in first immune defense system. In this research, we switch IgA class to IgG by replacing Fc portion of IgA with Fc portion from IgG, which is both having activity against Dengue. The mRNA comes from previous research, derived from hybridoma, which is generate from fusion peripheral blood lymphocytes from Thai patients with secondary infection of Dengue with “SPYMEG” as fusion partner. Stably expression system of Chinese Hamster Ovary (CHO)-K1 was generate for production of recombinant chimeric IgA-IgG. To purified recombinant chimeric IgA-IgG, single chromatography protein G shepharose was used and the productivity is 338 μg/ml. Kayword: Chimeric IgA-IgG, Monoclonal antibody, CHO-K1, purification, Dengue Virus

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Kata Kunci : Chimeric IgA-IgG, Monoclonal antibody, CHO-K1, purification, Dengue Virus