Immunological detection and quantitation of 10-deacetylbaccatin III in Taxus sp. plant and tissue cultures.
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.
Guo
Y
Y
Laboratoire de Pharmacognosie et de Bromatologie, Université Libre de Bruxelles, Belgique.
Jaziri
M
M
Diallo
B
B
Vanhaelen-Fastre
R
R
Zhiri
A
A
Vanhaelen
M
M
Homès
J
J
Bombardelli
E
E
D000918:Antibody Specificity; D001643:Bridged Bicyclo Compounds / Q000032:analysis / Q000276:immunology; D001952:Bridged-Ring Compounds; D002851:Chromatography, High Pressure Liquid; D003429:Cross Reactions; D046508:Culture Techniques; D004224:Diterpenes / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D010936:Plant Extracts / Q000032:analysis; D010946:Plants, Medicinal / Q000737:chemistry*; D013386:Succinates / Q000032:analysis / Q000737:chemistry; D043823:Taxoids; D014315:Triterpenes / Q000032:analysis* / Q000276:immunology / Q000378:metabolism
D001643:Bridged Bicyclo Compounds; D001952:Bridged-Ring Compounds; D004224:Diterpenes; D010936:Plant Extracts; D013386:Succinates; D043823:Taxoids; D014315:Triterpenes; C080625:taxane; C042213:10-deacetylbaccatine III