報告書番号

V10030

タイトル（和文）

ナノ・マイクロテクノロジーを利用した生物計測技術の開発(その１)- マイクロ流体デバイスを用いた重金属のイムノアッセイ-

タイトル（英文）

Development of biosensor using nano-micro technology (Part 1) -Immunoassay for heavy metals using microfluidic devices-

概要 （図表や脚注は「報告書全文」に掲載しております）

近年、さまざまな分野において重金属の簡易測定技術が求められており、従来の機器分析では困難な現場で測定しうる精度の高いイムノアッセイが望まれている。そこで、測定装置の小型化、測定時間の短縮や測定精度の改善が期待されるマイクロ流体デバイスを用いた重金属のイムノアッセイを構築した。微細加工技術により、樹脂製マイクロ流体デバイス（総容量1μL）を製作し、流路内に充填した担体粒子で起こる抗原抗体反応の度合いを吸光度から測定する検出部を設けた。マイクロ流体デバイスに対して抗カドミウム抗体、抗クロム抗体及び抗鉛抗体を適用し、流路への担体粒子の充填や試料流速・流量等の各種測定条件を最適化し、これらの諸条件を適切に組み合わせることで、抗原抗体反応と吸光度の関係から各重金属を環境基準値まで測定可能であることを証明した。また、単流路型流体デバイスの流路を集積化したマイクロ流体デバイスを作製し、抗カドミウム抗体を用いて四種類の試料のカドミウムの同時測定に成功した。集積型における測定感度は単流路型と同等であった。

概要 （英文）

A rapid and high sensitive measurement with low cost for heavy metals at in the field has been demanded to manage environment in food, product, waste and soil. Immunochromatography is thought to be a possible measurement technique in field ; however, its accuracy and sensitivity seem to be poor. At present, the application of microfabricated palmtop devices to conventional immunoassay has been focused on as one of strategies to improve the sensitivity and accuracy of measurements . Here, we describe working microfluidic devices, and we use these devices to perform immunoassay for heavy metals. Polydimethylsiloxane (PDMS) was casted on microfabricated templates to form a microchannel, and it was bonded to a glass slide. An electro-osmotic flow (EOF ) pump was employed to provide controllable and stable flow in the microchannel. Microbeads conjugated with a heavy metal were packed into the detection area of the microchannel; these microbeads were then coated with the free antibodies conjugated by gold colloid in the sample solution. The concentration of the antibodies coated on the microbeads was determined on the basis of absorbance. Monoclonal antibodies directed towards cadmium, chromium, and lead were used for microfluidic heavy metal immunoassays. Measurement conditions such as the concentration of antibodies, flow rate, and flow volume were optimized with measuring absorbance . Subsequently, the detection limits of each measurement satisfied the environmental standards for these heavy metal species. Moreover, four microchannels were integrated for the simultaneous and rapid measurement of four different samples. The sensitivity of measurement was equivalent to the single channel type

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