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Specific Protein Detection Using Designed DNA Carriers and Nanopores

2015; American Chemical Society; Volume: 137; Issue: 5 Linguagem: Inglês

10.1021/ja512521w

1943-2984

Nicholas A. W. Bell, Ulrich F. Keyser,

RNA Interference and Gene Delivery

Nanopores are a versatile technique for the detection and characterization of single molecules in solution. An ongoing challenge in the field is to find methods to selectively detect specific biomolecules. In this work we describe a new technique for sensing specific proteins using unmodified solid-state nanopores. We engineered a double strand of DNA by hybridizing nearly two hundred oligonucleotides to a linearized version of the m13mp18 virus genome. This engineered double strand, which we call a DNA carrier, allows positioning of protein binding sites at nanometer accurate intervals along its contour via DNA conjugation chemistry. We measure the ionic current signal of translocating DNA carriers as a function of the number of binding sites and show detection down to the single protein level. Furthermore, we use DNA carriers to develop an assay for identifying a single protein species within a protein mixture.