Optimization of Biosensors by Directed Evolution
Abstract
The objective of this work is to develop methodologies for the optimization of field-deployable optical biosensors. We used these Defense University Research Instrumentation Program funds to purchase a Perkin-Elmer GeneAmp PCR System 2400 thermal cycler and a SpectraMax Plus plate reader from Molecular Dynamics. We used the plate reader to develop faster assays for characterizing carbonic anhydrase (CA) variants. We used the thermal cycler to prepare a large library of CA variants. We then completed multiple rounds of selection for variants with enhanced zinc specificity using phage display. We successfully prepared variants with altered metal specificities using these methods. These variants can be used to optimize a carbonic anydrase-based metal ion biosensor.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jan 08, 2003
- Accession Number
- ADA410013
Entities
People
- Carol A. Fierke
Organizations
- University of Michigan