Abstract
The immobilization of glucose oxidase (GOx), using self assembled monolayers (SAMs) on gold surfaces, was investigated by grazing angle FT-IR spectroscopy, surface plasmon resonance (SPR) spectroscopy, and atomic force microscopy (AFM) in conjunction with confocal laser scanning microscopy (CLSM). To find an optimum condition for the maximum GOx loading density on gold surfaces, different cleaning protocols were examined. The loading density of GOx on surfaces was investigated by AFM and CLSM. In particular, CLSM was more effective for identifying the GOx density than AFM, since its scanning speed is much faster and it covers a larger area. Based on CLSM images of the GOx immobilized on the surfaces, it was concluded that the pre-cleaning process of gold substrates using different solvents, such as acetone, ethanol and 2-propanol, is very important for enhancing the GOx loading density. This result enables us to investigate an effective fabrication process in fabricating biosensors.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
E. Ostuni, L. Yan, and G. M. Whitesides, Colloids Surf., B, 1999, 15, 3.
H. O. Finklea, “Encyclopedia of Analytical Chemistry”, 2000, John Wiley & Sons Ltd., Chichester.
N. Patel, M. C. Davies, M. Hartshorne, R. J. Heaton, C. J. Roberts, S. J. B. Tendler, and P. M. Williams, Langmuir, 1993, 13, 6485.
J. J. Gooding, P. Erokhin, D. Losic, W. Yang, V. Policarpio, J. Liu, F. M. Ho, M. Situmorang, D. B. Hibbert, and J. G. Shapter, Anal. Sci., 2001, 17, 3.
D. Losic, J. G. Shapter, and J. J. Gooding, Langmuir, 2002, 18, 5422.
D. B. Hibbert, J. J. Gooding, and P. Erokhin, Langmuir, 2002, 18, 1770.
J. J. Gooding, L. Pugliano, D. B. Hibbert, and P. Erokhin, Electrochem. Commun., 2000, 2, 217.
A. J. Guiomar, J. T. Guthri, and S. D. Evans, Langmuir, 1999, 15, 1198.
J. J. Gooding, V. Praig, and E. A. H. Hall, Anal. Chem., 1998, 70, 2396.
J. J. Gooding, P. Erokhin, and D. B. Hibert, Biosens. Bioelectron., 2000, 15, 229.
K. T. Kinnear and H. G. Monbouquette, Langmuir, 1993, 9, 2255.
T. Lotzbeyer, W. Schuhmann, E. Katz, J. Falter, and H. L. Schmidt, J. Electroanal. Chem., 1994, 377, 291.
T. Lotzbeyer, W. Schuhmann, E. Katz, J. Falter, and H. L. Schmidt, Sens. Actuators, 1996, B33, 50.
T. Lotzbeyer, W. Schuhmann, E. Katz, J. Falter, and H. L. Schmidt, Bioelectroch. Bioener., 1997, 42, 1.
J. Li, G. Cheng, and S. Dong, J. Electroanal. Chem., 1996, 416, 97.
E. Katz, A. Riklin, V. Heleg-Shabtai, I. Willner, A. F. L. Buckmann, and A. Heller, J. Am. Chem. Soc., 1996, 118, 10132.
M. Mrksich and G. M. Whitesides, Ann. Rev. Biomol. Struct., 1996, 25, 55.
T. Wink, S. J. Van Zuilen, A. Bult, and W. P. Van Bennekom, Analyst, 1997, 122, 43R.
A. Ulman, “An Introduction to Ultrathin Organic Films; from Langmuir-Blodgett to Self-assembly”, 1991, Academic Press, San Diego.
C. D. Basin, J. Evall, and G. M. Whitesides, J. Am. Chem. Soc., 1989, 111, 7155.
K. E. Nelson, L. Gamble, L. S. Jung, M. S. Boeckl, E. Naeemi, S. L. Golledge, T. Sasaki, D. G. Castner, C. T. Campbell, and P. S. Stayton, Langmuir, 2001, 17, 2807.
N. Patel, M. C. Davis, M. Hartshorne, R. J. Heaton, C. J. Roberts, S. J. B. Tendler, and P. M. Williams, Langmuir, 1997, 13, 6485.
M. Veiseh, M. H. Zareie, and M. Zhang, Langmuir, 2002, 18, 6671.
M. Veiseh, Y. Zhang, K. Hinkley, and M. Zhang, Biomed. Microdev., 2001, 3, 43.
R. G. Chapman, E. Ostuni, S. Takayama, R. E. Holmlin, L. Yan, and G. Whitesides, J. Am. Chem. Soc., 2000, 122, 8303.
Author information
Authors and Affiliations
Corresponding authors
Rights and permissions
About this article
Cite this article
Park, T., Choo, J., Lee, M. et al. Enhancement of the Protein Loading Density by a Pre-Cleaning Process of a Gold Substrate: Confocal Laser Scanning Microscopic Study. ANAL. SCI. 20, 1255–1258 (2004). https://doi.org/10.2116/analsci.20.1255
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.2116/analsci.20.1255