Characterization of polyclonal and monoclonal anti-taxol antibodies and measurement of taxol in serum

Anti-taxol antibodies were generated in the rabbit using a taxol-bovine serum albumin conjugate prepared from 2'-succinyltaxol using a mixed anhydride procedure. Immunization with 2'-succinyltaxol-bovine serum albumin gave rise to polyclonal anti-taxol antibodies. By a radioimmunoassay usi...

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Bibliographic Details
Published in:Cancer research (Chicago, Ill.) Ill.), 1993-03, Vol.53 (6), p.1388-1391
Main Authors: JYH-GANG LEU, BI-XING CHEN, SCHIFF, P. B, ERLANGER, B. F
Format: Article
Language:English
Subjects:
Analysis
Animals
Antibodies - immunology
Antibodies, Monoclonal - immunology
Biological and medical sciences
Enzyme-Linked Immunosorbent Assay
Female
General pharmacology
Humans
Medical sciences
Paclitaxel - blood
Paclitaxel - chemistry
Paclitaxel - immunology
Pharmacology. Drug treatments
Rabbits
Radioimmunoassay
Structure-Activity Relationship
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Summary:Anti-taxol antibodies were generated in the rabbit using a taxol-bovine serum albumin conjugate prepared from 2'-succinyltaxol using a mixed anhydride procedure. Immunization with 2'-succinyltaxol-bovine serum albumin gave rise to polyclonal anti-taxol antibodies. By a radioimmunoassay using [3H]taxol, a standard curve gave a 50% inhibitory concentration of 1.0 nM. Taxol levels in human serum could be measured, with the lower limit of detection and measurement being 0.1 nM or 0.085 ng/ml. Two mouse monoclonal anti-taxol antibodies were isolated by immunizing BALB/c mice with the same antigen. One was an immunoglobulin G1 (69E4A8E) and the other was immunoglobulin M (29B7B3C). The specificity of these antibodies was determined by a competitive enzyme-linked immunosorbent assay with taxol and 10 different related derivatives and analogues. 29B7B3C had higher binding affinities for biologically active derivatives and markedly lower affinities for inactive derivatives; i.e., the specificity was consistent with the results of tubulin disassembly and cytotoxicity studies using the same taxol derivatives, making it suitable for screening for taxol or taxol-like compounds in extracts of natural products. 69E4A8E recognized the benzamidocarbamyl group at the C-3' position of taxol and had a lower affinity for other active compounds with different substitutions. Taxol levels in human serum could be detected and measured by 69E4A8E using a competitive enzyme-linked immunosorbent assay. The lower limit of measurement was about 50 nM or approximately 42 ng/ml. Similar measurements could be made by radioimmunoassay.
ISSN:0008-5472
1538-7445