Issue 1, 2010
From the journal:

Lab on a Chip

Micro OS-ELISA: Rapid noncompetitive detection of a small biomarkerpeptide by open-sandwich enzyme-linked immunosorbent assay (OS-ELISA) integrated into microfluidic device†

Masaki Ihara,‡ae   Amane Yoshikawa,b   Yushu Wu,b   Hiroko Takahashi,d   Kazuma Mawatari,cd   Kiyohito Shimura,ce   Kae Sato,ce   Takehiko Kitamoricde  and  Hiroshi Ueda*abe  

* Corresponding authors

a Department of Bioengineering, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo, Tokyo, Japan
E-mail: hueda@chembio.t.u-tokyo.ac.jp
Fax: +81-3-5841-7362

b Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo, Tokyo, Japan

c Department of Applied Chemistry, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo, Tokyo, Japan

d Microchemistry group, Kanagawa Academy of Science and Technology, 3-2-1 Sakado, Takatsu, Kawasaki, Kanagawa, Japan

e Center for NanoBio Integration, The University of Tokyo, 7-3-1 Hongo, Bunkyo, Tokyo, Japan

Abstract

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 (∼12 min), we could attain a detection limit comparable to that of the microplate-based OS ELISA (1 μg 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.

Graphical abstract: 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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Article information

DOI
https://doi.org/10.1039/B915516C
Article type
Paper
Submitted
29 Jul 2009
Accepted
30 Sep 2009
First published
03 Nov 2009

Lab Chip, 2010,10, 92-100

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Micro OS-ELISA: Rapid noncompetitive detection of a small biomarker peptide by open-sandwich enzyme-linked immunosorbent assay (OS-ELISA) integrated into microfluidic device

M. Ihara, A. Yoshikawa, Y. Wu, H. Takahashi, K. Mawatari, K. Shimura, K. Sato, T. Kitamori and H. Ueda, Lab Chip, 2010, 10, 92 DOI: 10.1039/B915516C

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