An Indirect Homologous Competitive Enzyme-linked Immunosorbent Assay for the Detection of a Class of Glycosylated Dihydrochalcones

Details

• Alternative Title:
  J Agric Food Chem
• Personal Author:
  Ranganathan, Anupama ; Paradise, Grace A. ; Hansen, Chad A. ; McCoy, Mark R. ; Gee, Shirley J. ; Zhong, Ping ; Chang, Dan ; Hammock, Bruce D.
• Description:
  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.
• Subjects:
  Animals Antibodies Antibody Specificity Article Dihydrochalcone Glycoside ELISA Enzyme-Linked Immunosorbent Assay Female Fermentation Flavanones Glucosides Hapten Haptens Immune Sera Immunization Microbial Fermentation Phloretin Polyclonal Antibodies Rabbits Serum Albumin, Bovine Taste

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• Source:
  J Agric Food Chem. 61(28):6964-6970
• Pubmed ID:
  23767873
• Pubmed Central ID:
  PMC3723731
• Document Type:
  Journal Article
• Funding:
  P42 ES004699/ES/NIEHS NIH HHS/United States ; U50 OH007550/OH/NIOSH CDC HHS/United States ; U54 OH007550/OH/NIOSH CDC HHS/United States ; OH07550/OH/NIOSH CDC HHS/United States
• Volume:
  61
• Issue:
  28
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:c947a72c10555a1a9c3e34323a0e58a6d63452f11fbc4615d6be907d31554e1a
• Download URL:
  https://stacks.cdc.gov/view/cdc/29692/cdc_29692_DS1.pdf
• File Type:
  [PDF - 694.69 KB ]