A competitive ELISA for quantification of protein a in culture medium


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

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Item Type: Article
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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