Cloning of recombinant fab from monoclonal antibody anti-dengue NS1 induced by recombinant CHO-K1 cells into pGEM-T vector

Abstract

Dengue infection, which is an endemic disease in tropical and subtropical countries, is commonly caused by one of the four serotypes of dengue virus. Indonesia as a tropical country has a high case of dengue infection. The dengue virus infection is usually manifested in a fever, but in severe conditions dengue infections are fatal. In order to minimize fatality rate of dengue infection, early diagnostic tests need to be developed. In this study, we aim to produce recombinant Fab from monoclonal antibody anti-Dengue NS1 (44F) induced by recombinant CHO-K1 cells into pGEM-T vector. The Light Chain (LC) of Fab coded by cDNA isolated from hybridoma cell 44F was amplified using PCR. The LC fragment was cloned into pGEM-T vector and transformed into E. coli TOP 10. Several transformants were selected for screening and confirmed by PCR and restriction enzyme digestion. The results showed that the LC gene sized 650 bp was successfully cloned into pGEM-T vector. Further study should be conducted in order to evaluate the level expression of the light chain gene of the Fab fragment as valuable material in the development of dengue diagnostic kit.