Development of an immunoassay using an anti-wogonin glucuronide monoclonal antibody

J Immunoassay Immunochem. 2017;38(5):457-470. doi: 10.1080/15321819.2016.1273236. Epub 2016 Dec 27.

Abstract

Wogonin 7-O-β-D-glucuronide (Wgn) is a bioactive flavone present in the dried root of Scutellaria baicalensis Georgi. To generate a monoclonal antibody (MAb) against Wgn, BALB/c mice injected with Wgn-bovine serum albumin yielded splenocytes that we fused with SP2/0 myeloma cells using the polyethylene glycol method. We obtained a hybridoma designated 315A that produced a MAb reactive to Wgn. The anti-Wgn MAb 315A was applied to an indirect competitive enzyme-linked immunosorbent assay (icELISA) to quantify Wgn. Subsequent validation revealed that icELISA using the 315A anti-Wgn MAb is an accurate and reliable method for the quantification of Wgn in S. baicalensis.

Keywords: Scutellaria baicalensis; Wogonin 7-O-β-D-glucuronide; indirect competitive enzyme-linked immunosorbent assay; monoclonal antibody.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology*
  • Antigen-Antibody Reactions
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Flavanones / analysis
  • Flavanones / immunology*
  • Glucuronides / analysis
  • Glucuronides / immunology*
  • Mice
  • Mice, Inbred BALB C

Substances

  • Antibodies, Monoclonal
  • Flavanones
  • Glucuronides
  • wogonin-7-glucuronide