Immunoaffinity chromatography for the sample pretreatment of Taxus plant and cell extracts prior to analysis of taxanes by high-performance liquid chromatography.
The application of immunoaffinity chromatography for the purification of Taxus plant and cell extracts prior to the HPLC analysis is described. Polyclonal antibodies raised against 10-deacetylbaccatin III (10-DAB III), paclitaxel's main precursor in plant, were characterised by enzymed-linked immunosorbent assay. Immunoglobulins from selected antisera were immobilised on CNBr-activated Sepharose 4B. The immunoaffinity column was used for the purification of plant and plant cell culture extracts prior to their analysis by HPLC. Immunoaffinity chromatography enabled the selective concentration of taxoids and enhanced sample clean-up.
Theodoridis
G
G
Laboratory of Analytical Chemistry, Dep. Chemistry, Aristotle University Thessaloniki, 540 06 Thessaloniki, Greece. gtheodor@chem.auth.gr
Haasnoot
W
W
Cazemier
G
G
Schilt
R
R
Jaziri
M
M
Diallo
B
B
Papadoyannis
I N
IN
de Jong
G J
GJ
D000906:Antibodies / Q000737:chemistry; D000972:Antineoplastic Agents, Phytogenic / Q000032:analysis / Q000302:isolation & purification; D001952:Bridged-Ring Compounds / Q000032:analysis*; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D004797:Enzyme-Linked Immunosorbent Assay; D007120:Immunochemistry; D018796:Immunoconjugates / Q000737:chemistry; D007074:Immunoglobulin G / Q000032:analysis; D010047:Ovalbumin / Q000737:chemistry; D017239:Paclitaxel / Q000032:analysis / Q000302:isolation & purification; D010936:Plant Extracts / Q000032:analysis; D012685:Sepharose; D012710:Serum Albumin, Bovine / Q000737:chemistry; D012997:Solvents; D013056:Spectrophotometry, Ultraviolet; D043823:Taxoids; D020946:Taxus / Q000737:chemistry* / Q000166:cytology; D014315:Triterpenes / Q000737:chemistry / Q000276:immunology
D000906:Antibodies; D000972:Antineoplastic Agents, Phytogenic; D001952:Bridged-Ring Compounds; D018796:Immunoconjugates; D007074:Immunoglobulin G; D010936:Plant Extracts; D012997:Solvents; D043823:Taxoids; D014315:Triterpenes; C080625:taxane; D012710:Serum Albumin, Bovine; C042213:10-deacetylbaccatine III; D010047:Ovalbumin; D012685:Sepharose; D017239:Paclitaxel