A Receptor Protein-Based Bioassay for Quantitative Determination of Paclitaxel

A novel receptor-based bioassay for the quantitative measurement of Taxol was developed. The assay was based on the well-investigated and established finding that Taxol, its active analogs, and active metabolites bind reversibly to the receptor protein tubulin, a process similar to antibody and anti...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 1997-09, Vol.69 (17), p.3633-3635
Main Authors: Suye, Shin-Ichiro, Tandel, Sagun, Mulchandani, Ashok
Format: Article
Language:English
Subjects:
Analysis
Animals
Antineoplastic Agents, Phytogenic - analysis
Biological and medical sciences
Cattle
Chemistry
Drugs
General pharmacology
Humans
Medical sciences
Paclitaxel - analysis
Pharmacology
Pharmacology. Drug treatments
Protein Binding
Receptors, Drug - chemistry
Receptors, Drug - isolation & purification
Tubulin - chemistry
Tubulin - isolation & purification
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Summary:A novel receptor-based bioassay for the quantitative measurement of Taxol was developed. The assay was based on the well-investigated and established finding that Taxol, its active analogs, and active metabolites bind reversibly to the receptor protein tubulin, a process similar to antibody and antigen interaction. The assay was performed in a competitive format by allowing a mixture of horseradish peroxidase-labeled Taxol and Taxol in the analyte sample to compete for the Taxol binding site of a polystyrene microtiter plate wall coated with purified tubulin and subsequently measuring the tubulin−Taxol complex by determining the activity of the horseradish peroxidase label. Using this method, Taxol was measured very sensitively, linear range of 0.0001−1 nM, and selectively, without interference from non-tumor-active compounds such as baccatin III, cephalomaninne, and 10-deacetyl taxol. The method was applied for the determination of picomolar concentrations of Taxol in human plasma.
ISSN:0003-2700
1520-6882
DOI:10.1021/ac970233h