Specific Protein Detection Using Designed DNA Carriers and Nanopores
Journal of the American Chemical Society
American Chemical Society
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Bell, N., & Keyser, U. (2015). Specific Protein Detection Using Designed DNA Carriers and Nanopores. Journal of the American Chemical Society, 137 2035-2041. https://doi.org/10.1021/ja512521w
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.
We thank Vivek Thacker and Nadanai Laohakunakorn for critical reading of this manuscript. N.A.W.B. was supported by an EPSRC Doctoral Prize Award. U.F.K. acknowledges support by an ERC starting grant, PassMembrane 261101.
European Research Council (261101)
External DOI: https://doi.org/10.1021/ja512521w
This record's URL: https://www.repository.cam.ac.uk/handle/1810/247474
Attribution 2.0 UK: England & Wales
Licence URL: http://creativecommons.org/licenses/by/2.0/uk/
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