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Fabrication of protein chips based on 3-aminopropyltriethoxysilane as a monolayer

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Abstract

Although 3-aminopropyltriethoxysilane (APTES) is widely adopted as a monolayer in biosensors, experimental silanization takes at least 1 h at high temperature. Therefore, the feasibility of the silanization with APTES in a short reaction time and at room temperature was investigated. The surface modification of glass slides using a self-assembled monolayer of APTES with a concentration of 10% was studied by immobilizing FITC. APTES was successfully immobilized on the glass slide. The effect of reaction temperature and time of silanization were investigated. Various silanization conditions of APTES were examined by contact angle measurement and fluorescence microscopy. The surface of glass patterns with a gold thin film as background was characterized by determining the fluorescent intensities following the immobilization of fluorescein isothiocyanate (FITC), protein A-FITC, antimouse IgG-FITC and sheep anti-bovine albumin-FITC. The normalized fluorescent intensity indicated that a short period (4 min) of silanization at 25°C suffices to form an APTES thin film by the immobilization of protein A on a glass surface. Such a condition does not require microheaters and temperature sensors in a microfluidic system, which will significantly reduce the manufacturing process, cost, and reaction time in the future.

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Acknowledgements

The authors would like to thank the National Science Council of Taiwan, the Republic of China, for financially supporting this research under Contract No. NSC 95-2622-E-006-039-CC3 and NSC 94-2218-E-006-043. The authors also would like to thank the Center for Micro/Nano Science and Technology, National Cheng Kung University, Tainan, Taiwan, for access to equipment and technical support. Furthermore, this work made use of Shared Facilities supported by the Program of Top 100 Universities Advancement, Ministry of Education, Taiwan.

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Correspondence to Ling-Sheng Jang.

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Jang, LS., Liu, HJ. Fabrication of protein chips based on 3-aminopropyltriethoxysilane as a monolayer. Biomed Microdevices 11, 331–338 (2009). https://doi.org/10.1007/s10544-008-9239-7

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