Development of an indirect competitive enzyme-linked immunosorbent assay (icELISA) using highly specific monoclonal antibody against paclitaxel

Zhi Chao, Mingming Tan, Madan Kumar Paudel, Seiichi Sakamoto, Liling Ma, Kaori Sasaki-Tabata, Hiroyuki Tanaka, Yukihiro Shoyama, Lijiang Xuan, Satoshi Morimoto

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Paclitaxel, the major active component of the yew tree, is used as an important anti-cancer agent. To obtain the monoclonal antibody (MAb) against paclitaxel for paclitaxel determination using immunoassay, 7-xylosyltaxol was conjugated to the carrier protein bovine serum albumin (BSA) to construct the immunogen, and the ratio of hapten in XylTax-BSA conjugate was determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. After immunization of mice with this conjugate, hybridomas secreting MAbs against paclitaxel were obtained by fusing the splenocytes with the mouse myeloma cell line SP2/0. After hybridoma screening, the anti-paclitaxel MAb 3A3 was obtained, which showed a relatively high specificity to paclitaxel (cross-reactivities against other naturally occurred taxanes: 7-xylosyltaxol, 31.8 %; cephalomannine, 6.17 %; baccatin III, 10-deacetyl-baccatin III, 1-hydroxybaccatin I, 13-acetyl-9-dihydrobaccatin III and 1-acetoxyl-5-deacetyl- baccatin I, <0.11 %). Using the MAb 3A3, we established an indirect competitive enzyme-linked immunosorbent assay (icELISA) for paclitaxel determination with a detection range of 0.098-312.5 μg ml-1. Determination of paclitaxel contents in various yew tree samples with this icELISA resulted in recovery rates ranging from 92 to 94.8 %, and intra- and inter-assay variations of 3.6 and 4.7 %, respectively. This icELISA provides a valuable method of paclitaxel determination for various purposes.

Original languageEnglish
Pages (from-to)512-518
Number of pages7
JournalJournal of Natural Medicines
Volume67
Issue number3
DOIs
Publication statusPublished - Jul 1 2013

All Science Journal Classification (ASJC) codes

  • Pharmaceutical Science
  • Drug Discovery
  • Complementary and alternative medicine
  • Organic Chemistry

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