Purification of NS2B-NS3 dengue virus serotype 3 protein as raw material for dengue virus vaccine

Abstract

Dengue virus serotype 3 has been recognized as the predominant serotype in many occurrences of Dengue Hemorrhagic Fever (DHF) in Indonesia. Performing DHF prevention could be done with vaccination. BPPT institution was developing a vaccine, which is made from NS2B-NS3 recombinant protein. This protein is one of the non-structural proteins that arrange the genome of DENV and it has a molecular weight of 83 kDa. This research aims to isolate and purify NS2B-NS3 protein DENV-3 from transformant cell of Saccharomyces cerevisiae. Purification of NS2B-NS3 protein is done with HisPur Ni-NTA Magnetic Beads method. Optimization of purification is done by increasing the concentration of imidazole as protein binder in the elution buffer starting from 250 mM until 500 mM. The validity of purified protein was tested qualitatively with Sodium Dodecyl Sulfate-Polyacrilamide Gel Electrophoresis (SDS-PAGE) method and quantified by Bichinconinic Acid (BCA) method. The results revealed us that NS2B-NS3 protein was purified optimally at 300 mM imidazole by HisPur Ni-NTA Magnetic Beads method. SDS-PAGE analysis showed that there was a specific band with size 83 kDa in lane result of elution with 300 mM imidazole and based on the result of protein quantification obtained that it has the highest percentage of purification effectiveness is 16.38 %.