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Proceedings Paper

Improved response of a fluorescence-based metal ion biosensor using engineered carbonic anhydrase variants
Author(s): Richard B. Thompson; Hui-Hui Zeng; Michele Loetz; Keith McCall; Carol A. Fierke
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Paper Abstract

The response time of biosensors which reversibly bind an analyte such as a metal ion is necessarily limited by the kinetics with which the biosensor transducer binds the analyte. In the case of the carbonic anhydrase-based biosensor we have developed the binding kinetics are rather slow, with the wild type human enzyme exhibiting an association rate constant ten thousand-fold slower than diffusion-controlled. By designed and combinatorial means the transducer may be mutagenized to achieve nearly diffusion-controlled association rate constants, with commensurate improvement in response. In addition, a variant of apocarbonic anhydrase has been immobilized on quartz, and is shown to response rapidly to changes in free copper ion in the picomolar range.

Paper Details

Date Published: 15 December 1999
PDF: 6 pages
Proc. SPIE 3858, Advanced Materials and Optical Systems for Chemical and Biological Detection, (15 December 1999); doi: 10.1117/12.372913
Show Author Affiliations
Richard B. Thompson, Univ. of Maryland/Baltimore (United States)
Hui-Hui Zeng, Univ. of Maryland/Baltimore (United States)
Michele Loetz, Univ. of Maryland/Baltimore (United States)
Keith McCall, Univ. of Michigan (United States)
Carol A. Fierke, Univ. of Michigan (United States)

Published in SPIE Proceedings Vol. 3858:
Advanced Materials and Optical Systems for Chemical and Biological Detection
Mahmoud Fallahi; Basil I. Swanson, Editor(s)

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