High-Throughput and High-Intensive Biosensor Microarray Fabrication by Selective Dewetting on a Wettability Controlled Substrate

In a microarray biosensor, the surface characteristic for reducing background nonspecific binding is a critical aspect for achieving the high sensitivity and accurate detection. In this paper, we proposed a new high-intensity and reduced nonspecific binding microarray fabrication method with a selec...

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Bibliographic Details
Published inJapanese Journal of Applied Physics Vol. 52; no. 1; pp. 017001 - 017001-6
Main Authors Kim, Jongsu, Park, Hyunkyu, Kang, Bongchul, Ham, Chulho, Lee, Hyeonhwa, Yang, Minyang
Format Journal Article
LanguageEnglish
Published The Japan Society of Applied Physics 01.01.2013
Subjects
Binding
Biosensors
Dewetting
Hydrophobicity
Proteins
Silanes
Surface properties
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Summary:In a microarray biosensor, the surface characteristic for reducing background nonspecific binding is a critical aspect for achieving the high sensitivity and accurate detection. In this paper, we proposed a new high-intensity and reduced nonspecific binding microarray fabrication method with a selective dewetting process on a mixed self-assembled monolayer (SAM), consisting of methyl-terminated and methoxy-[poly(ethylene glycol) (PEG)]-terminated silanes. The combination of hydrophobic and hydrophilic properties of the mixed SAM background layer provided the hydrophobicity that allowed the selective dewetting of the protein solution at specific versus nonspecific regions and reduced the nonspecific binding owing to hydrated PEG chains in the mixed SAM. The wiping process was applied for obtaining high throughput and reducing protein usage. Finally, the background non-specific binding was significantly reduced by 78% with a selective dewetting method compared with the standard method using a cover slip. Furthermore, the peak intensity was increased by 20% and the protein usage was reduced versus the standard method.
Bibliography:Concept of the proposed method: (a) Schematic process of microarray fabrication. (b) Wettability difference between the nonspecific binding region and the specific binding region. (c) Selective dewetting of protein solution. Wiping procedure on a patterned template (mixed OTS/PEG SAM, background layer; biotin-coated layer, specific region): (a) Schematic wiping process on mixed SAM-based microarray chip. (b) Selective dewetting of streptavidin solution on patterned biotin-coated region by wiping process. (c) Fluorescence array result for wiping speed under 2 mm/s. (d) Fluorescence array result for wiping speed over 2 mm/s. Protein (streptavidin) adsorption affected by wettability of substrate (mixing ratios of OTS/PEG SAM). Protein adsorption of the selective dewetting process on OTS layer and comparison with OTS and PEG layers prepared using the standard method. Contact angles of DI water on the mixed OTS/PEG SAM layer. Contact angles of DI water (25 °C), PBS buffer (25 °C), and 10 μg/mL streptavidin (2 °C) on the $1:1$ OTS/PEG SAM substrate. Background nonspecific binding comparison according to the OTS/PEG mixing ratio: standard method and selective dewetting method. Intensity profile and fluorescence image of the fabricated microarray: (a) Intensity profile of patterned protein as a $1:1$ mixed SAM background layer prepared by the selective dewetting method, and PEG layer prepared by the standard method. (b) Fluorescence image of Cy3-labeled streptavidin on the $1:1$ mixed SAM background layer prepared by the selective dewetting method. (c) Magnified image of 100 μm spot.
ObjectType-Article-1
SourceType-Scholarly Journals-1
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content type line 23
ISSN:0021-4922
1347-4065
DOI:10.7567/JJAP.52.017001