Nanotech 2011

Surface-Immobilization of Chromatographically Purified Bacteriophages for the Optimized Capture of Bacteria

R. Naidoo, A. Singh, S. Arya, B. Beadle, N. Glass, J. Tanha, C. Szymanski, S. Evoy
National Institute for Nanotechnology, University of Alberta, CA

Keywords: adsorption, bacteriophage, biosensor, chromatography, pathogen, virus immobilization

Abstract:

The removal of lysate contaminants from bacteriophage suspensions by size exclusion chromatography significantly increases the resultant planar surface density of immobilized bacteriophages. This leads to achieving maximum (jamming) surface coverage of phages and creates an opportunity to study the surface attachment isotherm more rigorously. Escherichia coli T4 and Salmonella enterica serovar Typhimurium P22 phage systems seem to undergo highly heterogeneous adsorption to the surface, possibly explaining the observed phage clustering at higher surface densities. The T4 phage and its E. coli host were initially employed as a model system where we discovered an optimal planar surface density of phages for best bacterial capture: 18.9 ± 0.8 phages/μm2 capturing 18.0 ± 0.3 bacteria/100 μm2. It is the phage surface clustering that ultimately limits the T4 phage-immobilized surface’s ability to specifically capture its host bacteria. Nevertheless, to our knowledge this is the largest surface capture density of E. coli reported using intact T4 bacteriophages. Two additional purified bacteriophage systems (P22 and Campylobacter jejuni phage NCTC 12673) were then similarly studied for their ability to capture their corresponding host bacteria (Salmonella enterica serovar Typhimurium and Campylobacter jejuni respectively) on a surface. More phage-bacteria pairs can be investigated, necessitating generalized methods for phage diversity.
 

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