Biomedical Microdevices

, Volume 11, Issue 2, pp 331–338 | Cite as

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



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.


Self-assembled monolayer APTES Protein chip 


  1. G. Arslan, M. Özmen, B. Gunduz, X. Zhang, M. Ersoz, Surface modification of glass beads with an aminosilane monolayer Turk. J. Chem. 30, 203–210 (2006)Google Scholar
  2. A.S. Blawas, W.M. Reichert, Protein patterning Biomaterials 19, 595–609 (1998)CrossRefGoogle Scholar
  3. T.L. Breen, J. Tien, S.R.J. Oliver, T. Hadzic, G.M. Whitesides, Self-assembly of mesoscale objects into ordered two-dimensional arrays Science 284, 948–951 (1999)CrossRefGoogle Scholar
  4. C. Ercole, M.D. Gallo, M. Pantalone, S. Santucci, L. Mosiello, C. Laconi, A. Lepidi, A biosensor for Escherichia coli based on a potentiometric alternating biosensing (PAB) transducer Sens. Actuators, B, Chem. 83(1–3), 48–52 (2002)CrossRefGoogle Scholar
  5. B.B. Haab, M.J. Dunham, P.O. Brown, Protein microarrays for highly parallel detection and quantitation of specific proteins and antibodies in complex solutions Genome Biol 2(4), 1–13 (2001)Google Scholar
  6. J.A. Howarter, J.P. Youngblood, Optimization of silica silanization by 3-aminopropyltriethoxysilane Langmuir. 22(26), 11142–11147 (2006)CrossRefGoogle Scholar
  7. B.H. Jo, L.M.V. Lerberghe, K.M. Motsegood, D.J. Beebe, Three-dimensional micro-channel fabrication in polydimethylsiloxane (PDMS) elastomer Journal of Microelectromechanical Systems 9, 76–81 (2000)CrossRefGoogle Scholar
  8. K.M.R. Kallury, R.F. DeBono, U.J. Krull, M. Thompson, ed. by K.L. Mittal. Silanes and other coupling agents (VSP, Zeist, Netherlands, 1992), pp. 263–269Google Scholar
  9. J.K. Kim, D. Shin, W.J. Chung, K.H. Jang, K.N. Lee, Y.K. Kim, Y.S. Lee, Effects of polymer grafting on a glass surface for protein chip applications Colloids Surf., B Biointerfaces. 33, 67–75 (2004)CrossRefGoogle Scholar
  10. W. Liao, X.T. Cui, Reagentless aptamer based impedance biosensor for monitoring a neuro-inflammatory cytokine PDGF Biosens. Bioelectron. 23, 218–224 (2007)CrossRefGoogle Scholar
  11. C.S. Liao, G.B. Lee, J.J. Wu, C.C. Chang, T.M. Hsieh, F.C. Huang, C.H. Luo, Micromachined Polymerase chain reaction system for multiple DNA amplification of upper respiratory tract infectious diseases Biosens. Bioelectron. 20, 1341–1348 (2005)CrossRefGoogle Scholar
  12. G. MacBeath, S.L. Schreiber, Printing proteins as microarrays for high-throughput function determination Science 289, 1760–1763 (2000)Google Scholar
  13. H. Neubert, E.S. Jacoby, S.S. Bansal, R.K. lles, D.A. Cowan, A.T. Kicman, Enhanced affinity capture MALDI-TOF MS: Orientation of an immunoglobulin G using recombinant protein G Anal. Chem. 74, 3677–3682 (2002)CrossRefGoogle Scholar
  14. P. Silberzan, L. Leger, D. Ausserre, J.J. Benattar, Silanation of silica surfaces. A new method of constructing pure or mixed monolayers Langmuir. 7, 1647–1651 (1991)CrossRefGoogle Scholar
  15. M.B. Stark, K. Holmberg, Covalent immobilization of lipase in organic solvents Biotechnol. Bioeng. 34, 942–950 (1989)CrossRefGoogle Scholar
  16. S. Susmel, C.K. O’Sullivan, G.G. Guilbault, Human cytomegalovirus detection by a quartz crystal microbalance immunosensor Enzyme Microb. Technol 1(27), 639–945 (2000)CrossRefGoogle Scholar
  17. T. Tanaka, T. Mastsunaga, Fully automated chemiluminescence immunoassay of insulin using antibody-protein a-bacterial magnetic particle complexes Anal. Chem. 72, 3518–3522 (2000)CrossRefGoogle Scholar
  18. A. Ulman, Formation and structure of self-assembled monolayers Chem. Rev. 96, 1533–1554 (1996)CrossRefGoogle Scholar
  19. K.D. Vos, I. Bartolozzi, E. Schacht, P. Bienstman, R. Baets, Silicon-on-insulator microring resonator for sensitive and label-free biosensing Optics Express 15(12), 7610–7615 (2007)CrossRefGoogle Scholar
  20. H. Yun, H. Bang, W.G. Lee, H. Lim, J. Park, J. Lee, A. Riaz, K. Cho, C. Chung, D.C. Han, J.K. Chang, Fluorescent intensity-based differential counting of FITC-doped silica nanoparticles: Applications of CD4+ T-cell detection in microchip-type flowcytometers Proc. Of SPIE. 6416, 641605–641612 (2007)CrossRefGoogle Scholar
  21. N.V. Zaytseva, V.N. Goral, R.A. Montagna, A.J. Baeumner, Development of a microfluidic biosensor module for pathogen detection Lab Chip 5, 805–811 (2005)CrossRefGoogle Scholar
  22. H. Zhu, M. Snyder, Protein arrays and microarrays Curr. Opin. Chem. Biol 5, 40–45 (2001)CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2008

Authors and Affiliations

  1. 1.Department of Electrical Engineering and Center for Micro/Nano Science and TechnologyNational Cheng Kung UniversityTainanTaiwan
  2. 2.TainanTaiwan

Personalised recommendations