Preparation and application of a monoclonal antibody against the isoflavone glycoside daidzin using a mannich reaction‐derived hapten conjugate. Issue 1 (January 2016)

• Record Type: Journal Article
• Authors: Yusakul, Gorawit; Sakamoto, Seiichi; Juengwatanatrakul, Thaweesak; Putalun, Waraporn; Tanaka, Hiroyuki; Morimoto, Satoshi
• Is Part Of: Phytochemical analysis. Volume 27:Issue 1(2016:Jan.)
• Page Start: 81
• Page End: 88
• Publication Date: 2016-01
• Subjects: ELISA -- monoclonal antibody -- daidzin, soybean -- Glycine max -- Pueraria lobata -- Pueraria mirifica; Plants -- Analysis -- Periodicals; Plants -- chemistry -- Periodicals; 572.2
• DOI: 10.1002/pca.2604
• Languages: English
• ISSNs: 0958-0344

Abstract

Abstract: Introduction: Daidzin and its aglycone daidzein are major pharmacologically active compounds of soybean ( Glycine max ), kudzu ( Pueraria lobata ), and kwao kruea khao ( P . mirifica ). Pharmacological activities of daidzin are mediated by its more potent metabolites daidzein and equol; however, daidzin is the predominant compound found in these medicinal plants, and the efficacy and safety of equol depend on the amount of daidzin consumed. Objective: To develop a specific monoclonal antibody (MAb)‐based indirect competitive enzyme‐linked immunosorbent assay (icELISA) for standardisation of daidzin content in herbal medicines or nutraceuticals. Methodology: The Mannich reaction was used for the synthesis of a highly immunogenic conjugate between daidzin and a cationised carrier protein. Splenocytes of hyperimmunised mice were fused with myeloma cells to generate a hybridoma secreting antibody against daidzin. Results: The icELISA showed high selectivity and acceptable sensitivity for daidzin determination (1.56–100 ng/mL) with high reproducibility (coefficients of variation were < 5%). The icELISA was a reliable analytical method for daidzin in Glycine max, Pueraria lobata and P . mirifica, for which daidzin recoveries from spiked samples were 98.99–104.94%. Daidzin content of these plant‐derived products determined using the icELISA were in close agreement with those determined by a HPLC‐UV method. Conclusion: The icELISA is useful for specific daidzin determination because of its reliability, low cost, speed and high throughput. Copyright © 2015 John Wiley & Sons, Ltd. Abstract : The Mannich reaction was used to synthesize cBSA‐daidzin conjugate for production of monoclonal antibody against daidzin. The icELISA showed high selectivity and acceptable sensitivity for daidzin determination (1.56–100 ng/mL) with high reproducibility. The icELISA was a reliable analytical method for daidzin in Glycine max, Pueraria lobata and Pueraria mirifica, for which daidzin recoveries from spiked samples were 98.99%–104.94%. Daidzin content of these plant‐derived products determined using the icELISA were in close agreement with a HPLC‐UV method.