Abstract
Aminosilane-treated molecular layers on glass surfaces are frequently used as functional platforms for biosensor preparation. All the amino groups present on the surface are not available in reactive forms, because surface amino groups interact with remaining unreacted surface silanol groups. Such nonspecific interactions might reduce the efficiency of chemical immobilization of biomolecules such as DNA, enzymes, antibodies, etc., in biosensor fabrication. To improve immobilization efficiency we have used additional surface silanization with alkylsilane (capping) to convert the remaining silanol groups into Si–O–Si linkages, thereby liberating the amino groups from nonspecific interaction with the silanol groups. We prepared different types of capped amine surface and evaluated the effect of capping on immobilization efficiency by investigating the fluorescence intensity of Cy3-NHS (N-hydroxysuccinimide) dye that reacted with amino groups. The results indicate that most of the capped amine surfaces resulted in enhanced efficiency of immobilization of Cy3-NHS compared with the untreated control amine surface. We found a trend that trialkoxysilanes had greater capping effects on immobilization efficiency than monoalkoxysilanes. It was also found that the aliphatic chain of alkylsilane, which does not participate in the capping of the silanol, had an important function in enhancing immobilization efficiency. These results would be useful for preparation of an amine-modified surface platform, with enhanced immobilization efficiency, which is essential for developing many kinds of biosensors on a silica matrix.
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Acknowledgements
This work is partially supported by Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science, and Technology, Japan, to K.M. (no. 18350083) and by Grants-in-Aid for Regional Science and Technology Promotion “Kyoto Nanotechnology Cluster” project from the Ministry of Education, Culture, Sports, Science and Technology, Japan. This Work was also supported by CREST of the Japan Science and Technology Agency. We also thank Yasuko Yoshida and Kazunari Yamada (NGK Insulators, Ltd, Geneshot Project, Japan) for providing the ink jet spotter and the image scanner for acquiring quantitative fluorescence data.
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Kamisetty, N.K., Pack, S.P., Nonogawa, M. et al. Development of an efficient amine-functionalized glass platform by additional silanization treatment with alkylsilane. Anal Bioanal Chem 386, 1649–1655 (2006). https://doi.org/10.1007/s00216-006-0741-6
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DOI: https://doi.org/10.1007/s00216-006-0741-6