Micro OS-ELISA: Rapid noncompetitive detection of a small biomarker peptide by open-sandwich enzyme-linked immunosorbent assay (OS-ELISA) integrated into microfluidic device
Micro OS-ELISA: Rapid noncompetitive detection of a small biomarker peptide by open-sandwich enzyme-linked immunosorbent assay (OS-ELISA) integrated into microfluidic device
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DOI：
10.1039/b915516c
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发表时间：
2010-01-01
期刊：
影响因子：
6.1
通讯作者：
Ueda, Hiroshi
中科院分区：
文献类型：
--
作者：
Ihara, Masaki;Yoshikawa, Amane;Ueda, Hiroshi
提出了一种新型检测系统，结合了开放夹心（OS）酶联免疫吸附测定（ELISA）和微流控传感器芯片系统的优点，能够对分子量小于1000的小抗原进行快速、非竞争性免疫检测。配备灵敏的热透镜显微镜，利用抗体可变区的抗原依赖性稳定性（OS原理）对微量人骨钙素（BGP）（骨代谢生物标志物）的羧基末端肽进行定量。在很短的分析时间内（大约 12 分钟），我们可以获得与基于微孔板的 OS ELISA (1 mu g L-1) 相当的检测限。此外，还研究了几种预处理对血清来源样品的影响：白蛋白吸收树脂、添加蛋白酶抑制剂混合物和热处理。发现每种预处理都是有效的。因此，与传统的竞争性 ELISA 方法相比，可以在健康人血清中检测到内在的 BGP 及其片段，并具有优异的检测限和工作范围。
A novel detection system that combines the merits of open-sandwich (OS) enzyme-linked immunoadsorbent assay (ELISA) and a microfluidic sensor chip system, and which enables rapid and noncompetitive immunodetection of small antigens of less than 1000 in molecular weight, has been proposed. Equipped with a sensitive thermal lens microscope, a minute amount of the carboxyl-terminal peptide of human osteocalcin (BGP), a biomarker for bone metabolism, was quantified utilizing antigen-dependent stabilization of an antibody variable region (OS principle). In a short analysis time (similar to 12 min), we could attain a detection limit comparable to that of the microplate-based OS ELISA (1 mu g L-1). In addition, the effects of several pretreatments for serum-derived samples were investigated: an albumin absorption resin, addition of a protease inhibitor cocktail and heat treatment. Each pretreatment was found to be effective. Consequently, intrinsic BGP and its fragments could be detected in healthy human serum with a superior detection limit and working range compared to those of the conventional competitive ELISA method.