Abstract
High-resolution patterning methods have been developed to immobilize functional proteins onto a silicon dioxide surface for biosensor applications. Antibody lines, as small as 5 μm in width, with intervening 5 μm spacings, were patterned on oxidized silicon wafers using avidin-biotin chemistry. The N-hydroxysuccinimide (NHS) ester of photoactivatable biotin was covalently bound to a self assembled monolayer (SAM) of 3-amino-propyltriethoxysilane (3-APTS) after irradiation by 350 nm ultraviolet (UV) light from a 25 W Hg arc lamp. The patterned layers were evaluated using fluorescent imaging of Alexa-488 conjugated avidin and two fluorescence-conjugated antibodies. This technique allows binding of any biotinylated compound without exposure to harmful UV light, extreme pH, toxic chemicals, or high salinity.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
R. Bashir, R. Gomez, A. Sarikaya, M.R. Ladisch, J. Sturgis, and J.P. Robinson, Biotechnology and Bioengineering 73, 324 (2001).
G. Bidan, M. Billon, K. Galasso, T. Livache, G. Mathis, A. Toget, L.M. Torres-Rodriguez, and E. Vieil, Appl. Biochem. Biotechnol. 89, 183 (2000).
J.D. Brewster, A.G. Gehring, R.S. Mazenko, L.J. Van Houten, C.J. Crawford, Anal. Chem. 68, 4153 (1996).
C.S.M. Chen, S. Huang, G. Whitesides, and D. Ingber, Biotechnol. Prog. 14, 356 (1998).
Z. Chnghua, P. Pivarnik, A.G. Rand, and S.V. Letcher, Biosens. Bioelectronics 15, 495 (1998).
N. Dontha, W.B. Nowall, and W.G. Kuhr, Anal. Chem. 69, 2619 (1997).
A.C. Forster, J.L. McInns, D.C. Skingle, and R. Symons, Nucleic Acid Res. 13, 745 (1985).
N.M. Green, Advances in Protein Chemistry 29, 85 (1975).
M. Hengsakul and A.E.G. Cass, Bioconj. Chem. 7, 249 (1996).
E. Lacey and W.N. Grant, Anal. Biochem. 163, 151 (1987).
M.C. Pirrung, and C.Y. Huang, Bioconjugate Chem. 7, 317 (1996).
D. Savage, G. Mattson, S. Desai, G. Niedlander, S. Morgensen, and E. Conklin, in Avidin-Biotin Chemistry: A Handbook, p. 76 (Pierce Chemical Company, Rockford, 1994).
P.M. St. John, R. Davis, N. Cady, J. Czajka, C.A. Batt, and H.G. Craighead, Anal Chem. 70, 1108 (1998).
T. Tsuchiya, in CRC Handbook of Organic Photochemistry and Photobiology, p. 980 (CRC Press, Boca Raton, 1995).
M. Wilchek, and E.A. Bayer, Methods in Enzymology 184, 174 (1990).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Orth, R.N., Clark, T. & Craighead, H. Avidin-Biotin Micropatterning Methods for Biosensor Applications. Biomedical Microdevices 5, 29–34 (2003). https://doi.org/10.1023/A:1024459215654
Issue Date:
DOI: https://doi.org/10.1023/A:1024459215654