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Bioconjugation of Meldrum’s acid Activated Furan: A Detergent Compatible Assay for Protein Quantitation

Publication Type : Journal Article

Publisher : Analytical Biochemistry

Source : Analytical Biochemistry 662 (2023) 114998,

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Campus : Amritapuri

School : School of Biotechnology

Year : 2023

Abstract : A simple yet efficient assay for the quantitation of proteins ranging from plasma proteins to purified proteins from whole cell lysate, based on the bioconjugation reaction between protein and Meldrum's acid Activated Furan (MAF) is described. This easy to use, sensitive method is based on the conjugation of amine functionalities present on the protein with MAF to form the corresponding Donor Acceptor Stenhouse Adducts (DASAs) with characteristic absorption in the visible region. The reaction is rapid as well as reproducible and shows a proportionate increase in color change over a broad range of protein concentration. The assay was found to be sensitive up to 0.125 mg/mL concentration of the protein and was compatible with most of the commonly employed detergents and isolation protocols which makes it ideal for the estimation of protein samples containing detergents. Another striking feature of this protocol is its tolerance towards other major interference contributors such as chelating agents, reducing agents, carbohydrates and protease inhibitors.

Cite this Research Publication : Bioconjugation of Meldrum’s acid activated furan: A detergent compatible assay for protein quantitation, Kalyani Ajayan, Sainath S, Ajmal Sadik, Manu Mohanan Nair, Anju M. Nair, Karthika K. S, Anagha Vijayakumar, Sudarslal Sadasivan Nair, Bipin Nair, Prakash Chandran R, Sobha Vijayan Nair ,Analytical Biochemistry 662 (2023) 114998,

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