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

bacteriophage immobilization, biotin binding polypeptide, cellulose binding module

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