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

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TechConnect Summit
Clean Technology 2008

Phage microarray

K.A. Vaglenov, S.N. Ustinov, G.A. Kuzmicheva, V.A. Petrenko
Auburn University, US

Keywords:
immunoassays, protein microarrays, antibodies, phage microarray, landscape bacteriophages, specificity, sensitivity, selectivity, high-throughput

Abstract:
A variety of immunoassays and protein microarrays have been developed recently as complementary to nucleic acid based microarrays to study genomes and proteomes of pathogens and macroorganisms with different pathologies. In these methods antibodies play a key role as unique and indispensable tool. Nevertheless, traditional methods of preparing antibodies are expensive, cumbersome and labor intensive. We developed alternative variant of protein microarray, which is based on landscape phage particles immobilized on an epoxy coated glass slides. Landscape phages are nanoparticles with regular structure displaying on their surface up to 4000 foreign peptide ligands. Employing two streptavidin binding phages as a model we demonstrated that phage-based probes can target low concentrated analytes with high sensitivity and discriminative selectivity. Method of phage immobilization on the epoxy-coated glass slide has been developed. Specificity of the arrays was evaluated with fluorescently labeled streptavidin and the sensitivity limit was estimated as ~ 1.0 nM. Selectivity was monitored in the presence of up to 14 molar excess of non-labeled streptavidin and up to 4000 molar excess of BSA. The system we described here is potentially capable to screen complex protein samples in high-throughput way using inexpensive available interfaces compared to the limited number of samples analyzed by traditional immunoassays such as ELISA.


Nanotech 2008 Conference Program Abstract