Capillary-based enzyme-linked immunosorbent assay for highly sensitive detection of thrombin-cleaved osteopontin in plasma

被引：22

作者：

Funano, Shun-ichi ^{[1
]}

Henares, Terence G. ^{[1
]}

Kurata, Mie ^{[2
,3
]}

Sueyoshi, Kenji ^{[1
]}

Endo, Tatsuro ^{[1
]}

Hisamoto, Hideaki ^{[1
]}

机构：

[1] Osaka Prefecture Univ, Grad Sch Engn, Dept Appl Chem, Osaka 5998531, Japan

[2] Ehime Univ, Grad Sch Med, Dept Pathogenom, Matsuyama, Ehime 7910295, Japan

[3] Erasmus MC, Ctr Thorax, Dept Cardiol, NL-3000 CA Rotterdam, Netherlands

来源：

ANALYTICAL BIOCHEMISTRY | 2013年 / 440卷 / 02期

关键词：

Square glass capillary; Enzyme-linked immunosorbent assay (ELISA); Thrombin-cleaved osteopontin (trOPN); ASSEMBLED MICROCHIP; SINGLE-STEP; INTEGRATION; SYSTEM; ANTIBODY; SERUM;

D O I：

10.1016/j.ab.2013.05.021

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

In this study, a highly sensitive capillary-based enzyme-linked immunosorbent assay (ELISA) has been developed for the analysis of picomolar levels of thrombin-cleaved osteopontin (trOPN), a potential biomarker for ischemic stroke, in human plasma. Using a square capillary coated with 8.5 mu g/ml anti-human trOPN capture antibody for ELISA, the linear range obtained was 2 to 16 pM trOPN antigen. This concentration range was in the detection window of trOPN antigen in plasma samples. Compared with the conventional microplate-based ELISA, the current capillary technique significantly reduced the amounts of reagent from milliliter to microliter, reduced the analysis time from 8 to 3 h, and had a better sensitivity and detection limit performance from approximately 50 pM down to 2 pM of trOPN antigen. These results indicate that this capillary-based immunoassay is a potential tool for biomarker detection and may be useful in clinical trials and medical diagnostic applications. (C) 2013 Elsevier Inc. All rights reserved.

引用

页码：137 / 141

页数：5

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