Purification of Fim-C-Salmonella typhi Recombinant Protein with Ni-NTA Resin as Raw Material for Typhoid Disease Detection Kit M Nurjayadi1., T Setiyoto1., S F Jinan1., D Hardianto2., A Sulfianti2., K Agustini2., HA El-Enshasy3
1 Department of Chemistry, Faculty of Mathematics and Natural Science, Universitas Negeri Jakarta Gedung KH. Hasjim Asj’ari, 6th Floor, Jl. Rawamangun Muka Jakarta Timur, 13220, Indonesia
2 Biofarmaka Research Laboratory LABTIAP BPPT-Serpong, Tangerang Selatan, Banten, Indonesia
3Institute of Bioproduct Development, Universiti Teknologi Malaysia (UTM), Skudai, Johor Bahru, Malaysia
Abstract
Typhoid caused by Salmonella typhi is an endemic disease in Asia, including Indonesia. Based on data from the World Health Organization (WHO), there are 81% per 100,000 people in Indonesia infected with typhoid. Based on this, development of typhoid detection kit that is simple, fast, specific and sensitive is still needed. One of the developments of detection kits currently available is the antigen and antibody interaction approach. This study aims to obtain comprehensive information about purification of Fim-C-S. typhi recombinant proteins as antigens for raw materials for manufacturing typhoid detection kits. This study uses the IMAC method with Ni2+ metal, which has advantages in high selectivity and purity. The results of this study report that purification of recombinant Fim-C-S. typhi protein inclusion bodies with variations in the binding and washing process at 3, 5, and 7 times gave percent yields correspondingly of 37.84, 39.44, and 38.21% pure protein. Previous studies have also reported variations in binding and washing at 2, 4, and 6 times and gave percent yields respectively were 35.37, 39.11, and 40.49%. Based on the data, we concluded that the variation of 6× is the best repetition variation to get the largest percent yield so that it can be used as a reference in large-scale purification.
Keywords: Salmonella typhi, Purification Protein, Recombinant Fim-C S. typhi Protein, IMAC, Ni- NTA
