A Competitive ELISA for Quantification of Protein a in Culture Medium
Ali, Abdul Manaf and Tahir, Sharifah and Abdul Ghani, Baharuddin and Ungku Mohsin, Ungku Chulan and Ahmad, Ismail (1996) A Competitive ELISA for Quantification of Protein a in Culture Medium. Pertanika Journal of Tropical Agricultural Science, 19 ((2/3)). pp. 95-102. ISSN 0126-6128
A competitive enzyme-linked immunosorbent assay (ELISA) for quantification of protein A produced by Staphylococcus aureus A676 (a methicillin-resistant strain) was based on competitive binding to rabbit IgG molecules between alkaline phosphatase-labelled protein A and unlabelled protein A. The optimum IgG concentration required for coating was 2-4 ug/ml. The optimum incubation time for colour development using (p-nitrophenol phosphate substrate was 20-30 min. The lowest protein A concentration that could be measured using the optimized competitive ELISA was 20 ng/ml, and the maximum 2 Ug/ml. The amount of protein A produced in brain-heart infusion medium increased exponentially during log phase of cell growth, reaching a maximum concentration of 22.5 Ug/ml after 15 h cultivation.
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