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 |
| 巻 | 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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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
}
TY - JOUR
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.
UR - http://www.scopus.com/inward/record.url?scp=84879239893&partnerID=8YFLogxK
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U2 - 10.1007/s11418-012-0708-1
DO - 10.1007/s11418-012-0708-1
M3 - Article
C2 - 23007175
AN - SCOPUS:84879239893
VL - 67
SP - 512
EP - 518
JO - Journal of Natural Medicines
JF - Journal of Natural Medicines
SN - 1340-3443
IS - 3
ER -