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Proceedings Paper

Enhanced biosensor performance using an avidin-biotin bridge for antibody immobilization
Author(s): Upvan Narang; George P. Anderson; Keeley D. King; Heidi S. Liss; Frances S. Ligler
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Paper Abstract

Maintaining antibody function after immobilization is critical to the performance of a biosensor. The conventional methods to immobilize antibodies onto surfaces are via covalent attachment using a crosslinker or by adsorption. Often, these methods of immobilization result in partial denaturation of the antibody and conformational changes leading to a reduced activity of the antibody. In this paper, we report on the immobilization of antibodies onto the surface of an optical fiber through an avidin-biotin bridge for the detection of ricin, ovalbumin, and Bacillus globigii (Bg). The assays are performed in a sandwich format. First, a capture antibody is immobilized, followed by the addition of the analyte. Finally, a fluorophore- labeled antibody is added for the specific detection of the analyte. The evanescent wave-induced fluorescence is coupled back through the same fiber to be detected using a photodiode. In all cases, we observe an improved performance of the biosensor, i.e., lower limit of detection and wide linear dynamic range, for the assays in which the antibody is immobilized via avidin-biotin bridges compared to covalent attachment method.

Paper Details

Date Published: 7 May 1997
PDF: 8 pages
Proc. SPIE 2980, Advances in Fluorescence Sensing Technology III, (7 May 1997); doi: 10.1117/12.273534
Show Author Affiliations
Upvan Narang, Naval Research Lab. (United States)
George P. Anderson, Naval Research Lab. (United States)
Keeley D. King, Naval Research Lab. (United States)
Heidi S. Liss, Naval Research Lab. (United States)
Frances S. Ligler, Naval Research Lab. (United States)

Published in SPIE Proceedings Vol. 2980:
Advances in Fluorescence Sensing Technology III
Richard B. Thompson, Editor(s)

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