Indirect homologous competitive enzyme-linked immunosorbent assay for the detection of a class of glycosylated dihydrochalcones

Anupama Ranganathan, Grace A. Paradise, Chad A. Hansen, Mark R. McCoy, Shirley J. Gee, Ping Zhong, Dan Chang, Bruce D. Hammock

Research output: Contribution to journalArticle

3 Scopus citations

Abstract

Hesperetin dihydrochalcone 4′-glucoside, 1, and phloretin 4′-glucoside, 2, belong to a family of dihydrochalcone glycosides that exhibit flavorant properties. In this study was developed a competitive, indirect homologous ELISA for the detection of targets 1 and 2 in fermentation media. Immunogen and coating antigen were prepared by conjugating hapten, 4-(3-oxo-3-(2,6-dihydroxy-4-glucoside phenyl)propyl) benzoic acid, to thyroglobulin and bovine serum albumin, respectively. Antibodies raised in rabbits M6122, M6123, and M6124 and the coating antigen were screened and characterized to determine their optimum concentrations. The optimized ELISA, developed with antibody M6122, gave IC50 values of 27.8 and 21.8 ng/mL for 1 and 2, respectively. Selectivity of the assay was assessed by measuring cross-reactivity of antibody M6122 to related congeners such as aglycones and the 2′-glycosides of hesperetin dihydrochalcone, 5 and phloretin, 6. Antibody M6122 showed very low recognition of 5 and virtually no recognition of the aglycones and 6.

Original languageEnglish (US)
Pages (from-to)6964-6970
Number of pages7
JournalJournal of Agricultural and Food Chemistry
Volume61
Issue number28
DOIs
StatePublished - Jul 17 2013

Keywords

  • dihydrochalcone glycoside
  • ELISA
  • hapten
  • microbial fermentation
  • polyclonal antibodies

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Chemistry(all)

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    Ranganathan, A., Paradise, G. A., Hansen, C. A., McCoy, M. R., Gee, S. J., Zhong, P., Chang, D., & Hammock, B. D. (2013). Indirect homologous competitive enzyme-linked immunosorbent assay for the detection of a class of glycosylated dihydrochalcones. Journal of Agricultural and Food Chemistry, 61(28), 6964-6970. https://doi.org/10.1021/jf401302y