Systems, apparatuses and methods for reading an amino acid sequence

Patent metadata

Field	Value
Publication number	US-10139417-B2
Application number	US-201314376154-A
Country	US
Kind code	B2
Filing date	Jan 31, 2013
Priority date	Feb 1, 2012
Publication date	Nov 27, 2018
Grant date	Nov 27, 2018

Abstract

Embodiments of the present disclosure relate to amino acid, modified amino acid, peptide and protein identification and sequencing, by means of, for example, electronic detection of individual amino acids or small peptides.

First claim

What is claimed is:

1. An apparatus for sequencing a molecule comprising:
at least a first and a second electrode;
a dielectric layer of between about 1 nm and about 4 nm separating the first and second electrodes;
a membrane between about 10 to about 100 nm in thickness;
a channel containing an electrolyte solution, wherein the membrane spans and divides the channel to form a cis chamber and a trans chamber,
wherein the cis chamber and the trans chamber are in fluid communication with one another via a nanopore, and
a bead in the cis chamber and arranged within 150 μm of the nanopore, the bead including a molecule for sequencing and/or identifying bound thereto,
wherein:
the nanopore is formed on the same substrate as the pair of electrodes,
the nanopore is between about 1 nm and about 5 nm in diameter at the point where it passes through the electrodes, and
the electrodes include between about 1 nm to about 5 nm of silver, gold, palladium and/or platinum, and the electrodes are functionalized.

2. The apparatus according to claim 1 , wherein each chamber is filled with an electrolyte comprising at least one of KCl and NaClO3 in a concentration between about 1 mM to about 1M.

3. The apparatus according to claim 1 , wherein the bead includes enzymes attached thereto which are fixed in turn to the walls of the channel.

4. The apparatus according to claim 3 , wherein the enzymes comprise proteases selected from the group consisting of trypsin, chymotrypsin, pepsin, papain, elastase and combinations thereof.

5. The apparatus according to claim 3 , wherein the bead is functionalized with proteosomes to sequentially degrade proteins into their component amino acids.

6. The apparatus according to claim 3 , wherein the molecule comprises an isolated protein or peptide.

7. The apparatus according to claim 1 , further comprising one or more reference electrodes.

8. The apparatus according to claim 1 , wherein the bead is affixed to the wall of the channel.

9. The apparatus according to claim 1 , wherein the molecule is not a protease.

Assignees

• Arizona Board Of Regents Acting For And On Behalf Of Arizona State Univ
• Univ Arizona State

Inventors

• Lindsay Stuart
• Zhang Peiming
• Zhao Yanan

Classifications

• G01N33/6821 Primary

  involving C-terminal degradation · CPC title

• G01N33/48721 Primary

  Investigating individual macromolecules, e.g. by translocation through nanopores (Coulter counters in general G01N15/12; fabrication methods for nanoscale apertures B81B1/00; sequencing of nucleic acids C12Q1/68) · CPC title