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Proceedings Paper

Photoactivatable silanes for the site-specific immobilization of antibodies
Author(s): David W. Conrad; Anna V. Davis; Sara K. Golightley; John C. Bart; Frances S. Ligler
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Paper Abstract

The development of techniques which can be used for the discrete immobilization of multiple antibodies on solid supports has important applications in the areas of clinical diagnostics, drug discovery, and biosensor development. While photolithography has provided many elegant methods for generating spatially resolved reactive surface functionalities, the problem of nonspecific adsorption of proteins which occurs during sequential immobilization steps continues to hamper the development of antibody sensing arrays. This nonspecific adsorption prevents the establishment of unique and individually addressable binding sites on the substrate surface. In an effort to solve this problem, we have synthesized and characterized a series of photoactivatable silanes which can be used for the light- directed attachment of antibodies to solid supports. These compounds produce amine-reactive functional groups upon irradiation with UV light. The first silane derivative prepared yielded a relatively hydrophobic surface when it was used for substrate modification. The resultant surface exhibited a high degree of nonspecific protein adsorption even in the absence of light activation. We therefore decided to synthesize a more hydrophilic analog. This second derivative displayed low nonspecific protein binding even in the absence of detergents or protein blocking agents. We have used this second compound to discretely immobilize four different IgG antibodies (each directed against a different antigen) on several hydroxyl-bearing substrates. Antigen binding experiments using fluorescent and 125I-labeled antigens have confirmed that the immobilized antibodies retain their functionalities.

Paper Details

Date Published: 31 March 1997
PDF: 10 pages
Proc. SPIE 2978, Micro- and Nanofabricated Electro-Optical Mechanical Systems for Biomedical and Environmental Applications, (31 March 1997); doi: 10.1117/12.269962
Show Author Affiliations
David W. Conrad, Naval Research Lab. (United States)
Anna V. Davis, GEO-CENTERS, Inc. (United States)
Sara K. Golightley, GEO-CENTERS, Inc. (United States)
John C. Bart, American Society for Engineering Education (United States)
Frances S. Ligler, Naval Research Lab. (United States)

Published in SPIE Proceedings Vol. 2978:
Micro- and Nanofabricated Electro-Optical Mechanical Systems for Biomedical and Environmental Applications
Paul Lee Gourley, Editor(s)

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