Photoactivatable silanes for the site-specific immobilization of antibodies

David W. Conrad; Anna V. Davis; Sara K. Golightley; John C. Bart; Frances S. Ligler

doi:10.1117/12.269962

31 March 1997 Photoactivatable silanes for the site-specific immobilization of antibodies

David W. Conrad, Anna V. Davis, Sara K. Golightley, John C. Bart, Frances S. Ligler

Proceedings Volume 2978, Micro- and Nanofabricated Electro-Optical Mechanical Systems for Biomedical and Environmental Applications; (1997) https://doi.org/10.1117/12.269962
Event: BiOS '97, Part of Photonics West, 1997, San Jose, CA, United States

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.

Citation Download Citation

David W. Conrad, Anna V. Davis, Sara K. Golightley, John C. Bart, and Frances S. Ligler "Photoactivatable silanes for the site-specific immobilization of antibodies", Proc. SPIE 2978, Micro- and Nanofabricated Electro-Optical Mechanical Systems for Biomedical and Environmental Applications, (31 March 1997); https://doi.org/10.1117/12.269962

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