2008 NSTI Nanotechnology Conference and Trade Show - Nanotech 2008 - 11th Annual

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TechConnect Summit
Clean Technology 2008

Engineering of bacteriophages displaying affinity tags on its head for biosensor applications

M. Tolba, L.Y. Brovko and M.W. Griffiths
University of Guelph, CA

Keywords:
bacteriophage immobilization, biotin binding polypeptide, cellulose binding module

Abstract:
A method was developed for the introduction of affinity tags onto bacteriophage heads using a phage display technique. It was shown that T4 bacteriophage carrying a biotin-binding polypeptide (Biotin Carboxyl Carrier Protein, BCCP, from acetyl-CoA carboxylase of E. coli) or a Cellulose Binding Module (CBM, of xylanase from Thermotoga maritime) can be effectively immobilized onto various streptavidin- or cellulose-based materials, respectively. Interaction of the bacteriophage with the functionalized surface is strong and almost irreversible. The immobilized phages, termed BCCP-T4 and CBM-T4, retain a high level of infectivity and are capable of capturing 70-100% of E. coli cells from suspensions containing 10 to 106 cfu/ml in 30 min. The subsequent lysis of captured cells resulted in release of multiple progeny phages. Enumeration of progeny phages by real-time PCR allowed detection of 1 cfu of E. coli per 5 µl sample within 2 hours.


Nanotech 2008 Conference Program Abstract