Rapid Determination of Immunoglobulin A in Human Saliva by Enzyme-Linked Immunosorbent Assay on a Micro Channel Chip

• NAKAJIMA Hizuru

  Faculty of Urban Environmental Sciences, Tokyo Metropolitan University

• MASUDA Hironori

  Faculty of Urban Environmental Sciences, Tokyo Metropolitan University

• ISHINO Satomi

  Faculty of Urban Environmental Sciences, Tokyo Metropolitan University

• NAKAGAMA Tatsuro

  Faculty of Urban Environmental Sciences, Tokyo Metropolitan University

• SHIMOSAKA Takuya

  National Metrology Institute of Japan, National Institute of Advanced Industrial Science and Technology

• ARAI Kensuke

  School of Pharmacy, Nihon Pharmaceutical University

• YOSHIMURA Yoshihiro

  School of Pharmacy, Nihon Pharmaceutical University

• UCHIYAMA Katsumi

  Faculty of Urban Environmental Sciences, Tokyo Metropolitan University

Bibliographic Information

Other Title

• マイクロチャネルを用いる酵素免疫測定法によるヒト唾液中イムノグロブリンＡの迅速定量
• マイクロチャネル オ モチイル コウソ メンエキ ソクテイホウ ニ ヨル ヒト ダエキ チュウ イムノグロブリン A ノ ジンソク テイリョウ

Abstract

An enzyme-linked immunosorbent assay for the rapid determination of Immunoglobulin A (IgA) in human saliva has been developed on a microchannel chip. First, a primary antibody (anti-IgA) was adsorbed on the surface of a polydimethylsiloxane microchannel. The channel was then filled with an antigen (IgA) solution, followed by the introduction of a secondary antibody (anti-IgA HRP labeled) solution. Finally, a substrate solution (phosphate buffer containing Amplex^® Red and hydrogen peroxide) was introduced into the microchannel, and the enzyme turnover was monitored by a fluorescent imaging system equipped with a CCD camera. The calibration curve of IgA standard solution at the enzyme-substrate reaction time of 5 minutes was linear over the range of 0∼50 ng/ml with a correlation coefficient of 0.998. The former assay on a 96-well microtiter plate required 150 min for the determination of IgA, while the present assay on the microchannel chip enables the determination of IgA in less than 35 min. This method was successfully applied to the determination of IgA in human saliva. An on-line flow assay system with a microchannel chip has also been developed.<br>

Journal

• BUNSEKI KAGAKU

  BUNSEKI KAGAKU 54 (9), 817-823, 2005

  The Japan Society for Analytical Chemistry

Keywords

• microfluidics
• immunoglobulin A
• saliva
• stress

Details

• CRID: 1390282679332291200
• NII Article ID: 110002910488
• NII Book ID: AN00222633
• DOI: 10.2116/bunsekikagaku.54.817
• COI: 1:CAS:528:DC%2BD2MXhtVGis7bI
• NDL BIB ID: 7489230
• ISSN: 05251931
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I7489230
• Web Site: http://www.jstage.jst.go.jp/article/bunsekikagaku/54/9/54_9_817/_pdf
• Web Site: http://dl.ndl.go.jp/info:ndljp/pid/10896209
• Web Site: https://search.jamas.or.jp/link/ui/2006061059
• Text Lang: ja
• Data Source:
  • JaLC
  • NDL
  • Crossref
  • NDL-Digital
  • CiNii Articles
  • KAKEN
• Abstract License Flag: Disallowed