Label-free biomolecule sensor based on surface charge modulated ionic conductance
First Claim
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1. A method for detecting biological analytes comprising a nucleic acid sequence with a charged backbone using ionic conductance through one or more nano-sized pores, the method comprising:
- providing a nanoporous membrane substrate having one or more nano-sized pores;
covalently binding onto a wall of the one or more nano-sized pores a capturing element comprising a morpholino oligonucleotide and/or a peptide nucleic acid oligonucleotide, wherein the morpholino and/or PNA oligonucleotide comprises a neutral charged backbone structure;
exposing the capturing element that is covalently bound onto the wall of the one or more nano-sized pores to a solution possessing the biological analytes, wherein the biological analytes are complementary to the capturing element and require no modification so that biological complementary analytes specifically bind to the capturing element and alter the surface charge of the nanopore; and
exposing the one or more nano-sized pores bulk concentration of electrolytes low enough so that the surface charge on walls of the one or more nano-sized pores dictates the concentration of ions inside the one or more nano-sized pores and causes the ionic conductance to become independent of the bulk concentration of electrolytes;
low concentration of electrolytes;
detecting a change of ion concentration as a result of ions being carried through the one or more nano-sized pores being of opposite charge as the charge of the wall of the one or more nano-sized pores and thus realizing an increase of ionic conductance through the one or more nano-sized pores upon specific binding of the capturing element and the biological complementary analytes.
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Abstract
The present invention provides for the detection of biochemical analytes by measuring variations in ionic conductance resulting from the hybridization of a biological analyte (e.g. oligonucleotides) with a capturing element immobilized on a membrane substrate having nano-sized pores.
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Citations
6 Claims
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1. A method for detecting biological analytes comprising a nucleic acid sequence with a charged backbone using ionic conductance through one or more nano-sized pores, the method comprising:
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providing a nanoporous membrane substrate having one or more nano-sized pores; covalently binding onto a wall of the one or more nano-sized pores a capturing element comprising a morpholino oligonucleotide and/or a peptide nucleic acid oligonucleotide, wherein the morpholino and/or PNA oligonucleotide comprises a neutral charged backbone structure; exposing the capturing element that is covalently bound onto the wall of the one or more nano-sized pores to a solution possessing the biological analytes, wherein the biological analytes are complementary to the capturing element and require no modification so that biological complementary analytes specifically bind to the capturing element and alter the surface charge of the nanopore; and exposing the one or more nano-sized pores bulk concentration of electrolytes low enough so that the surface charge on walls of the one or more nano-sized pores dictates the concentration of ions inside the one or more nano-sized pores and causes the ionic conductance to become independent of the bulk concentration of electrolytes;
low concentration of electrolytes;detecting a change of ion concentration as a result of ions being carried through the one or more nano-sized pores being of opposite charge as the charge of the wall of the one or more nano-sized pores and thus realizing an increase of ionic conductance through the one or more nano-sized pores upon specific binding of the capturing element and the biological complementary analytes. - View Dependent Claims (2, 3, 4, 5, 6)
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Specification