Purification of Salmonella Typhi Fim-C recombinant proteins with Co-NTA resins as raw materials for development of rapid typhoid fever detection kit

Abstract

The Fim-C-S typhi recombinant protein is a protein developed from the surface of the Salmonella typhi bacteria. This protein has been known to cause a good immune response. The protein has the potential as a raw material for immuno chromatography-based with gold nano particles as rapid detection kits. Various alternatives are taken to obtain pure protein with the maximum amount. Previous studies have succeeded in purifying the Fim-C-S. typhi recombinant protein with the Ni/NTA approach. This study aims to obtain information on optimum conditions for Fim-C-S. typhi recombinant protein purification with Co/NTA resin as raw material for rapid detection kit, because Co/NTA resin is also known to have good specifications for purification of recombinant proteins with Histidine tagging. There are three main stages in the purification process in this system, namely binding, washing, and elution. The binding and washing processes in this study were carried out 2, 4 and 6 times. Furthermore, the elution process was carried out at an imidazole concentration of 200 mM, 250 mM, and 300 mM. The results of the analysis using ImageJ gel analysis software on the results of the characterization using SDSPAGE showed that the purification in the binding process four and six times showed almost the same intensity of the band with size 31 KDa. The calculation of the yield of purification results showed the highest number of 42.75% in the six times washing process and imidazole concentration of 300 mM. Based on the data, it can be concluded purification of Fim-C-S. typhi recombinant proteins with Co-NTA resin give good results in the binding process four times, washing six times and 300 mM elution concentration. This purified Fim-C-S. typhi recombinant protein will then be used as an antigen in the development of a typhoid fever detection rapid kits.