15 December 1999 Improved response of a fluorescence-based metal ion biosensor using engineered carbonic anhydrase variants
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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.
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Richard B. Thompson, Hui-Hui Zeng, Michele Loetz, Keith McCall, 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); doi: 10.1117/12.372913; https://doi.org/10.1117/12.372913
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