A Competitive ELISA for quantification of protein a in culture medium

A Competitive ELISA for quantification of protein a in culture medium

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 opt...

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Bibliographic Details
Main Authors: Ali, Abdul Manaf, Tahir, Sharifah, Abdul Ghani, Baharuddin, Ungku Mohsin, Ungku Chulan, Ahmad, Ismail
Format: Article
Language:English
Published: Universiti Putra Malaysia Press 1996
Online Access:http://psasir.upm.edu.my/id/eprint/3521/1/A_Competitive_ELISA_for_Quantification_of_Protein_A_in_Culture_Mediunt.pdf
Description
Summary: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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