Production of monoclonal antibodies against a major purgative component, sennoside B, their characterization and use in ELISA

Analyst. 2001 Aug;126(8):1372-6. doi: 10.1039/b103153h.

Abstract

For immunization, sennoside B was conjugated with bovine serum albumin. The hapten density in the antigen conjugate was determined to be 3 mol mol(-1) protein by matrix-assisted laser desorption-ionization TOF mass spectrometry. A hybridoma secreting monoclonal antibody against sennoside B was produced by fusing splenocytes from mouse immunized with the sennoside B conjugate and mouse myeloma cells. Weak cross-reactivities occurred with sennoside A which is a stereochemical isomer, and a monomer of sennoside B, rhein, but no cross-reactivity was observed with other related anthraquinones and phenolics. The range of the assay extended from 0.5 ng ml(-1) to 15 ng ml(-1) of sennoside B, and good correlation between ELISA and HPLC methods was obtained when crude extracts of rhubarb were analyzed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anthraquinones / analysis
  • Anthraquinones / immunology*
  • Antibodies, Monoclonal / isolation & purification*
  • Cathartics / analysis*
  • Enzyme-Linked Immunosorbent Assay / methods
  • Senna Extract
  • Sennosides

Substances

  • Anthraquinones
  • Antibodies, Monoclonal
  • Cathartics
  • Sennosides
  • Senna Extract