Uun Yanuhar
Tuberculosis is a major infectious disease in the world until recently, but efforts to develop diagnostic tests based on antigen-antibody on antigen-antibody reaction to the present results are not encouraging. The purpose of this study was to develop antibodies adhesin Pili isolated from sputum of tuberculosis patients through hybridoma fusion. In this study, Mycobacterium tuberculosis (Mtb) Pili as an antigen and IgG antibody as secondary antibody for the diagnosis of Tb. Research method is Pili isolated with ultrahigh centrifugation at 120,000 rpm in a stratified at 4 {down triangleC and cuts with pili cuter (omnimixer). Molecular weight determined by electrophoresis, SDS Page (Sodium Dodecyl Sulfate polyacrylamide gel electrophoresis), Electroelution and produce antibodies by fusion between lymphocyte b cell and hybridoma cells that have been induced by adhesion protein pili of Mtb. The study show that pili adhesion proteins of Mtb as a hemagglutinin molecule 38 kDa is used for process production of monoclonal antibodies with form hybrid cells. Results of antibody testing produced from hybridoma cells showed that hybrid cell that contain protein of Mtb 38 kDa cross-reacted with the test sample of 96 sputum samples of TB patients using dot blot, ELISA and western blott. The conclusion is hybrid cell on the culture can results medium culture that reaction with protein adhesion pili of 38 kDa as epitope that binding with antibody that produced by hybridoma cells.
Water Sciences and Marine Biotechnology Laboratory, Brawijaya University, Malang, Indonesia
