Production of monoclonal antibodies against Staphylococcus aureus serotype 5

Abstract

"Staphylococcus aureus" infections are associated with a high mortality rate, as well as high morbidity and medical expense. Each year in the United States alone, they cause approximately 260,000 hospital acquired infections, with Serotype 5 and 8 Staphylococcus aureus (S.aureus) being the most prominent. Serotype 5 and 8 S. aureus are surrounded by a capsular polysaccharide which inhibits phagocytosis, thus preventing the body from ridding itself from the bacteria. In this study we have produced monoclonal antibodies to Type 5 and 8 S. aureus. Plasma cells (antibody producing cells) from the spleen of immunized mice were fused to a myeloma cell line to form hybridomas. These hybridomas were tested by Indirect ELISA for antibodies binding to purified capsular polysaccharide from S. aureus. Those that tested positive for binding were subcloned, tested for binding to the purified CP, and then for binding to whole bacteria using Indirect ELISA's. The positive binding subclones were subcloned a second time, and again tested for antigen binding. If the second subclones were positive for binding, they were frozen. These monoclonal antibodies should be valuable tools for the diagnosis and treatment of Staphylococcus aureus infections.