Molecular Approaches to Optical Biosensors
Abstract
The goal of this proposal was to develop methodologies for the optimization of field-deployable optical biosensors, in general, and, in particular, to optimize a carbonic anhydrase-based fiber optic zinc biosensor. We used both site-directed and random mutagenesis coupled with phage display to prepare and identify carbonic anhydrase (CA) variants that: (1) enhance the kinetics of metal >/= 1000-fold such that zinc equilibration in the nM range occurs within seconds; (2) alter the zinc affinity by a factor of 10(exp 7)-fold (20 fM - 0.2 micro-M) for use in a biosensor array; and (3) improve the metal detection limit by altering the transduction scheme. Additionally, we have developed phage display technology to screen for CA variants with alter zinc affinity.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jan 01, 1998
- Accession Number
- ADA365045
Entities
People
- Carol A. Fierke
Organizations
- Duke University Hospital