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Development of an enzyme-linked immunosorbent assay for the quantification of O-Phosphoethanolamine in human plasma

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:
2022年12月
DOI:
10.1016/j.ab.2022.114952
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:
*Tetsuya Saita, Hiroto Kataoka, Rintaro Sogawa, Tadashi Hayama, Ryoko Tomita, Akira Monji, Yoshito Mizoguchi, Chisato Shimanoe
題名:
Development of an enzyme-linked immunosorbent assay for the quantification of O-Phosphoethanolamine in human plasma
発表情報:
Anal Biochem 巻: 659 ページ: 114952
キーワード:
概要:
O-Phosphoethanolamine (PEA) is an endogenous substance that is attracting interest as a biomarker for depression, and thus there is a need to develop a simple analytical method that specifically measures PEA. Therefore, this study aimed to develop a simple and specific enzyme-linked immunosorbent assay (ELISA) for PEA. Anti-PEA antibody was obtained by immunizing mice with an antigen conjugated with mercaptosuccinyl bovine serum albumin using m-maleimidobenzoyl-N-hydroxysulfosuccinimide ester (MBS). In this assay, the PEA to be quantified is chemically modified by benzoyl chloride that is allowed to compete with a PEA-MBS-HRP conjugate for binding to a limited amount of an anti-PEA antibody, which was used to coat the wells of a microtiter plate. This ELISA shows a linear range of detection of 0.11-27 μM, and a limit of quantification of 0.144 μM. The anti-PEA antibody showed high affinity for benzoyl PEA. No detectable cross-reactivity was found with benzoyl 2-aminoethanol, O-phospho-l-tyrosine or benzoyl sphingosine-1-phosphate. The values of plasma PEA levels measured by this ELISA were comparable to those measured by HPLC, and a strong correlation was observed between the values determined by the two methods. The developed ELISA should provide a valuable new tool for the quantification of PEA in human plasma.
抄録:

英語フィールド

Author:
*Tetsuya Saita, Hiroto Kataoka, Rintaro Sogawa, Tadashi Hayama, Ryoko Tomita, Akira Monji, Yoshito Mizoguchi, Chisato Shimanoe
Title:
Development of an enzyme-linked immunosorbent assay for the quantification of O-Phosphoethanolamine in human plasma
Announcement information:
Anal Biochem Vol: 659 Page: 114952
An abstract:
O-Phosphoethanolamine (PEA) is an endogenous substance that is attracting interest as a biomarker for depression, and thus there is a need to develop a simple analytical method that specifically measures PEA. Therefore, this study aimed to develop a simple and specific enzyme-linked immunosorbent assay (ELISA) for PEA. Anti-PEA antibody was obtained by immunizing mice with an antigen conjugated with mercaptosuccinyl bovine serum albumin using m-maleimidobenzoyl-N-hydroxysulfosuccinimide ester (MBS). In this assay, the PEA to be quantified is chemically modified by benzoyl chloride that is allowed to compete with a PEA-MBS-HRP conjugate for binding to a limited amount of an anti-PEA antibody, which was used to coat the wells of a microtiter plate. This ELISA shows a linear range of detection of 0.11-27 μM, and a limit of quantification of 0.144 μM. The anti-PEA antibody showed high affinity for benzoyl PEA. No detectable cross-reactivity was found with benzoyl 2-aminoethanol, O-phospho-l-tyrosine or benzoyl sphingosine-1-phosphate. The values of plasma PEA levels measured by this ELISA were comparable to those measured by HPLC, and a strong correlation was observed between the values determined by the two methods. The developed ELISA should provide a valuable new tool for the quantification of PEA in human plasma.


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