Improved response of a fluorescence-based metal ion biosensor using engineered carbonic anhydrase variants

Richard B. Thompson; Hui-Hui Zeng; Michele Loetz; Keith McCall; Carol A. Fierke

doi:10.1117/12.372913

15 December 1999 Improved response of a fluorescence-based metal ion biosensor using engineered carbonic anhydrase variants

Richard B. Thompson, Hui-Hui Zeng, Michele Loetz, Keith McCall, Carol A. Fierke

Author Affiliations +

Proceedings Volume 3858, Advanced Materials and Optical Systems for Chemical and Biological Detection; (1999) https://doi.org/10.1117/12.372913
Event: Photonics East '99, 1999, Boston, MA, United States

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.

Citation Download Citation

Richard B. Thompson, Hui-Hui Zeng, Michele Loetz, Keith McCall, and Carol A. Fierke "Improved response of a fluorescence-based metal ion biosensor using engineered carbonic anhydrase variants", Proc. SPIE 3858, Advanced Materials and Optical Systems for Chemical and Biological Detection, (15 December 1999); https://doi.org/10.1117/12.372913

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