Citation
Baharuddin Abdul Ghani, . and Sharifah Tahir, . and Ungku Chulan, . and Ismail B. Ahmad, . and Abdul Manaf Ali, . A competitive ELISA for quantification of protein a in culture medium. pp. 95-102. ISSN 0126-6128
Abstract
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-labelledprotein A and unlabelledprotein A. The optimum IgG concentration required for coating was 2-4 fig/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 fig/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 fig/ml after 15 h cultivation.
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Abstract
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-labelledprotein A and unlabelledprotein A. The optimum IgG concentration required for coating was 2-4 fig/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 fig/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 fig/ml after 15 h cultivation.
Additional Metadata
Item Type: | Article |
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AGROVOC Term: | Proteins |
AGROVOC Term: | Enzyme linked immunosorbent assay |
AGROVOC Term: | Staphylococcus aureus |
AGROVOC Term: | Alkaline phosphatase |
AGROVOC Term: | Cultivation |
AGROVOC Term: | Optical density |
AGROVOC Term: | Purification |
Depositing User: | Ms. Suzila Mohamad Kasim |
Last Modified: | 24 Apr 2025 06:27 |
URI: | http://webagris.upm.edu.my/id/eprint/22722 |
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