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.

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Document Details

Document Type
Technical Report
Publication Date
Jan 08, 2003
Accession Number
ADA410013

Entities

People

  • Carol A. Fierke

Organizations

  • University of Michigan

Tags

Communities of Interest

  • Biomedical

DTIC Thesaurus Topics

  • Abstracts
  • Amino Acids
  • Biochemistry
  • Biosensors
  • Chemistry
  • Classification
  • Dynamic Range
  • Dynamics
  • Fluorescence
  • Instrumentation
  • Laboratory Equipment
  • Molecular Dynamics
  • Optimization
  • Personal Information Managers
  • Polymerase Chain Reaction
  • Thermal Cyclers
  • Universities

Readers

  • Molecular Genetics
  • Research Science/Academic Research
  • Thermal Physics or Thermal Science.

Technology Areas

  • Biotechnology