title: Indirect homologous competitive enzyme-linked immunosorbent assay for the detection of a class of glycosylated dihydrochalcones.
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.
authors.affiliation: Department of Entomology and UCD Comprehensive Cancer Center, University of California Davis, Davis, California 95616, United States.
authors.forename: Anupama
authors.initials: A
authors.lastname: Ranganathan
authors.forename: Grace A
authors.initials: GA
authors.lastname: Paradise
authors.forename: Chad A
authors.initials: CA
authors.lastname: Hansen
authors.forename: Mark R
authors.initials: MR
authors.lastname: McCoy
authors.forename: Shirley J
authors.initials: SJ
authors.lastname: Gee
authors.forename: Ping
authors.initials: P
authors.lastname: Zhong
authors.forename: Dan
authors.initials: D
authors.lastname: Chang
authors.forename: Bruce D
authors.initials: BD
authors.lastname: Hammock
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology; D000918:Antibody Specificity; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D005285:Fermentation; D044950:Flavanones / Q000032:analysis* / Q000276:immunology; D005960:Glucosides / Q000032:analysis* / Q000276:immunology; D006241:Haptens / Q000737:chemistry / Q000276:immunology; D007106:Immune Sera / Q000096:biosynthesis; D007114:Immunization; D010693:Phloretin / Q000031:analogs & derivatives* / Q000032:analysis / Q000276:immunology; D011817:Rabbits; D012710:Serum Albumin, Bovine / Q000276:immunology; D013649:Taste
chemical_list: D000906:Antibodies; D044950:Flavanones; D005960:Glucosides; D006241:Haptens; D007106:Immune Sera; D012710:Serum Albumin, Bovine; D010693:Phloretin