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Development of a sandwich enzyme-linked immunosorbent assay for the quantification of ponatinib in serum

発表形態:
原著論文
主要業績:
主要業績
単著・共著:
共著
発表年月:
2019年04月
DOI:
10.1016/j.ab.2019.02.010
会議属性:
指定なし
査読:
有り
リンク情報:

日本語フィールド

著者:
*Yuta Yamamoto, Tetsuya Saita, Rintaro Sogawa, Kenji Ogata, Yutaro Yamamoto, Sakiko Kimura, Yutaka Narisawa, Shinya Kimura, Masashi Shin
題名:
Development of a sandwich enzyme-linked immunosorbent assay for the quantification of ponatinib in serum
発表情報:
Anal Biochem 巻: 571 ページ: 14-20
キーワード:
概要:
The tyrosine kinase inhibitor ponatinib is extensively metabolized in the body, and consequently the development of specific immunoassays for pharmacokinetic studies and therapeutic drug monitoring of ponatinib is challenging. If two antibodies simultaneously recognize the entire structure of ponatinib, they could be utilized to establish an ultra-specific sandwich immunoassay for ponatinib, free of any interference from ponatinib metabolites. In this study, we created two types of anti-ponatinib polyclonal antibodies that recognize two different ponatinib epitopes, and sandwiched almost all structural components of ponatinib in these two antibodies in order to develop an enzyme-linked immunosorbent assay (ELISA) technique not affected by any ponatinib metabolites. After optimization, this sandwich ELISA showed a linear detection range of 640 pg/mL to 2000 ng/mL and a limit of quantification of 640 pg/mL. This sandwich ELISA was specific to ponatinib and showed no cross-reactivity with the major metabolite M14. Comparison between the sandwich ELISA and HPLC, using serum samples from 15 rats orally administered a single dose of 15 mg/kg ponatinib, showed a linear regression (y = 0.9662x + 3.5354, r = 0.9683). Thus, in this study, we successfully developed the first ultra-specific sandwich ELISA for ponatinib in serum.
抄録:

英語フィールド

Author:
*Yuta Yamamoto, Tetsuya Saita, Rintaro Sogawa, Kenji Ogata, Yutaro Yamamoto, Sakiko Kimura, Yutaka Narisawa, Shinya Kimura, Masashi Shin
Title:
Development of a sandwich enzyme-linked immunosorbent assay for the quantification of ponatinib in serum
Announcement information:
Anal Biochem Vol: 571 Page: 14-20
An abstract:
The tyrosine kinase inhibitor ponatinib is extensively metabolized in the body, and consequently the development of specific immunoassays for pharmacokinetic studies and therapeutic drug monitoring of ponatinib is challenging. If two antibodies simultaneously recognize the entire structure of ponatinib, they could be utilized to establish an ultra-specific sandwich immunoassay for ponatinib, free of any interference from ponatinib metabolites. In this study, we created two types of anti-ponatinib polyclonal antibodies that recognize two different ponatinib epitopes, and sandwiched almost all structural components of ponatinib in these two antibodies in order to develop an enzyme-linked immunosorbent assay (ELISA) technique not affected by any ponatinib metabolites. After optimization, this sandwich ELISA showed a linear detection range of 640 pg/mL to 2000 ng/mL and a limit of quantification of 640 pg/mL. This sandwich ELISA was specific to ponatinib and showed no cross-reactivity with the major metabolite M14. Comparison between the sandwich ELISA and HPLC, using serum samples from 15 rats orally administered a single dose of 15 mg/kg ponatinib, showed a linear regression (y = 0.9662x + 3.5354, r = 0.9683). Thus, in this study, we successfully developed the first ultra-specific sandwich ELISA for ponatinib in serum.


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