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 Tabata, Hiroyuki Tanaka, Yukihiro Shoyama, Lijiang Xuan, Satoshi Morimoto

研究成果: ジャーナルへの寄稿記事

14 引用 (Scopus)

抄録

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.

元の言語英語
ページ(範囲)512-518
ページ数7
ジャーナルJournal of Natural Medicines
67
発行部数3
DOI
出版物ステータス出版済み - 7 1 2013

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Immunosorbents
Paclitaxel
Assays
Enzyme-Linked Immunosorbent Assay
Monoclonal Antibodies
Enzymes
Hybridomas
Bovine Serum Albumin
Immunization
Taxoids
Haptens
Immunoassay
Ionization
Mass spectrometry
Desorption
Mass Spectrometry
Carrier Proteins
Screening
Lasers
Cells

All Science Journal Classification (ASJC) codes

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

これを引用

Development of an indirect competitive enzyme-linked immunosorbent assay (icELISA) using highly specific monoclonal antibody against paclitaxel. / Chao, Zhi; Tan, Mingming; Paudel, Madan Kumar; Sakamoto, Seiichi; Ma, Liling; Tabata, Kaori; Tanaka, Hiroyuki; Shoyama, Yukihiro; Xuan, Lijiang; Morimoto, Satoshi.

:: Journal of Natural Medicines, 巻 67, 番号 3, 01.07.2013, p. 512-518.

研究成果: ジャーナルへの寄稿記事

Chao, Zhi ; Tan, Mingming ; Paudel, Madan Kumar ; Sakamoto, Seiichi ; Ma, Liling ; Tabata, Kaori ; Tanaka, Hiroyuki ; Shoyama, Yukihiro ; Xuan, Lijiang ; Morimoto, Satoshi. / Development of an indirect competitive enzyme-linked immunosorbent assay (icELISA) using highly specific monoclonal antibody against paclitaxel. :: Journal of Natural Medicines. 2013 ; 巻 67, 番号 3. pp. 512-518.
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title = "Development of an indirect competitive enzyme-linked immunosorbent assay (icELISA) using highly specific monoclonal antibody against paclitaxel",
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.",
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T1 - Development of an indirect competitive enzyme-linked immunosorbent assay (icELISA) using highly specific monoclonal antibody against paclitaxel

AU - Chao, Zhi

AU - Tan, Mingming

AU - Paudel, Madan Kumar

AU - Sakamoto, Seiichi

AU - Ma, Liling

AU - Tabata, Kaori

AU - Tanaka, Hiroyuki

AU - Shoyama, Yukihiro

AU - Xuan, Lijiang

AU - Morimoto, Satoshi

PY - 2013/7/1

Y1 - 2013/7/1

N2 - 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.

AB - 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.

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