Enzyme-Linked Immunosorbent Assay Using Vertical Micro Reactor Stack for the Detection of Biomolecules

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  • 積層型マイクロリアクターを用いた酵素免疫測定法による生体物質測定系の開発
  • セキソウガタ マイクロリアクター オ モチイタ コウソ メンエキ ソクテイホウ ニ ヨル セイタイ ブッシツ ソクテイケイ ノ カイハツ

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Abstract

Microreactors and micro total analysis system (μTAS) are recognized as powerful tools for genomics, proteomics, clinical diagnostics, and environmental testing. In this paper, we describe enzyme linked immunosorvent assay (ELISA) using a new microreactor with a vertical fluid flow operation. This microreactor is composed of two reaction vessels stacked on the vertical lines through PMMA fluid filters (φ3mm). The fluid filters constructed by deep X-ray lithography possess 2,100 pores (φ 40 μm), and have valve functions, which maintain liquid layer in each reaction vessel. In addition, the liquid can be selectively transferred by air pressure from upper vessel to lower, and vice versa. As a model of ELISA using the microreactor, we planed to detect mouse immunoglobulin (IgG). We bound the goat anti-IgG antibody to the surface of the PMMA filters, and assayed the IgG by ELISA using anti-IgG antibody/ peroxidase conjugate. We found that the mouse IgG (100 ng/ml) was quantitatively detected within 45 min of analytical period, which was ca. 1/3 of the period required for the conventional method using micro titer plate.

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