Immunological detection and quantitation of 10-deacetylbaccatin III in Taxus sp. plant and tissue cultures

Biol Chem Hoppe Seyler. 1994 Apr;375(4):281-7. doi: 10.1515/bchm3.1994.375.4.281.

Abstract

A high-sensitive ELISA method was developed for the detection and semi-quantitative determination of 10-deacetylbaccatin III and its structurally related compounds in crude extract of Taxus sp. plants and tissue cultures. The antibodies were raised in rabbits using 7- or 10-succinyl-10-deacetylbaccatin III-BSA conjugate as immunogen. The working range of the assay was from 0.003 to 1.000 ng (0.09 to 31.33 nM) of 10-deacetylbaccatin III per assay. The cross-reacting material in crude plant extract was examined by chromatographic (silica gel CC, HPLC) and immunoassay methods. Study on the evaluation of cross-reacting material in crude Taxus plant extracts showed that at least 80% of the immunosignal correspond to 10-deacetylbaccatin III in the extract. The ELISA method was applied to investigate the 10-deacetylbaccatin III equivalent content in crude extracts of 19 plants species including Taxaceae, Taxodiaceae and Pinaceae species. The 10-deacetylbaccatin III-like structure was only detected in Taxus and Torreya sp. The results indicate that this immunoassay is a useful tool for the rapid screening of species, varieties or individual plants out of a wide population. The distribution of 10-deacetylbaccatin III equivalent content in 9-month old Taxus plantlets cultivated in vitro as well as in callus culture was investigated.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibody Specificity
  • Bridged Bicyclo Compounds / analysis
  • Bridged Bicyclo Compounds / immunology
  • Bridged-Ring Compounds*
  • Chromatography, High Pressure Liquid
  • Cross Reactions
  • Culture Techniques
  • Diterpenes / metabolism
  • Enzyme-Linked Immunosorbent Assay
  • Plant Extracts / analysis
  • Plants, Medicinal / chemistry*
  • Succinates / analysis
  • Succinates / chemistry
  • Taxoids*
  • Triterpenes / analysis*
  • Triterpenes / immunology
  • Triterpenes / metabolism

Substances

  • Bridged Bicyclo Compounds
  • Bridged-Ring Compounds
  • Diterpenes
  • Plant Extracts
  • Succinates
  • Taxoids
  • Triterpenes
  • taxane
  • 10-deacetylbaccatine III