US 9,791,453 B2Granted

Methods for determining binding capability of target ligands with G protein-coupled receptors using translocation through nanochannels

AssigneesInternational Business Machines Corporation, The University of Melbourne, The Florey Institute of Neuroscience and Mental Health

InventorsMatthew Downton, Natalie Gunn, Stefan Harrer, Priscilla Rogers, John Wagner, Ross Bathgate, Daniel Scott, Stan Skafidas

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Abstract

A mechanism is provided for utilizing a nanodevice to distinguish molecules with different structure. The molecules translocate through or across a nanochannel filled with a electrolyte solution. An electrical signal through the nanochannel is measured for every translocation event. Inner surfaces of the nanochannel include a functional layer, which is a coating to functionalize the nanochannel, in which the functional layer is configured to interact with predetermined ones of the molecules during translocation events. It is determined that a combination of at least two different molecules is formed based on predetermined ones of the molecules interacting with the functional layer to change the electrical signal and/or change a translocation time for the translocation event.

Claims27

Dependent claims

A method for determining binding capability of target ligands with G protein-coupled receptors in a test solution, the method comprising:

providing the G protein-coupled receptors each initially bound to a large tracer ligand, the large tracer ligand comprising albumin;
introducing the target ligands to the G protein-coupled receptors, wherein the target ligands are smaller than the large tracer ligand;
translocating the target ligands and the G protein-coupled receptors partially or entirely through nanochannels in parallel, the nanochannels filled with an electrolyte solution, the translocating is based on an electric potential difference applied by a voltage source in a longitudinal direction of the nanochannels to define an event, wherein the longitudinal direction is a length of the nanochannels;
measuring one or more electrical signals for every event, events being at least one translocation event, at least one binding event, or a combination of at least one translocation event and at least one binding event, the one or more electrical signals determined by at least one signal being measured through the nanochannels in the longitudinal direction, wherein measuring the one or more electrical signals comprises continuously measuring time and amplitude for the one or more electrical signals, wherein the one or more electrical signals correspond to respective ionic current through the nanochannels;
and
determining whether the target ligands and the G protein-coupled receptors are bound to one another based on a modulation in the one or more electrical signals to recognize a translocation time for the translocation event in one of the nanochannels;
in response to the large tracer ligand being bound to the G protein-coupled receptors while translocating the G protein-coupled receptors through the nanochannels, determining that the large tracer ligand remains bound to the G protein-coupled receptors due to a long time duration for the large tracer ligand bound to the G protein-coupled receptors traveling through the nanochannels, thereby recognizing that the target ligands did not replace the large tracer ligand;
in response to the large tracer ligand being displaced from being bound to the G protein-coupled receptors and in response to the large tracer ligand being replaced with the target ligand while translocating the G protein-coupled receptors through the nanochannels, determining that the target ligands are bound to the G protein-coupled receptors due to a short time duration for the target ligand bound to the G protein-coupled receptors traveling through the nanochannels, thereby recognizing that the target ligands have replaced the large tracer ligand;
wherein the short time duration of travel time in the nanochannels is shorter than the long time duration.

6 more elements

A method for determining binding capability of target ligands with investigative molecules in a test solution, the method comprising:

providing the investigative molecules initially bound to a large tracer ligand, the large tracer ligand comprising albumin;
introducing the target ligands to the investigative molecules, wherein the target ligands are smaller than the large tracer ligand;
translocating the target ligands and the investigative molecules partially or entirely through nanochannels in parallel, the nanochannels filled with an electrolyte solution, the translocating is based on an electric potential difference applied by a voltage source in a longitudinal direction of the nanochannels to define an event, wherein the longitudinal direction is a length of the nanochannels;
measuring one or more electrical signals for every event, events being at least one translocation event, at least one binding event, or a combination of at least one translocation event and at least one binding event, the one or more electrical signals determined by at least one signal being measured through the nanochannels in the longitudinal direction, wherein measuring the one or more electrical signals comprises continuously measuring time and amplitude for the one or more electrical signals, wherein the one or more electrical signals correspond to respective ionic current through the nanochannels;
and
determining whether the target ligands and the investigative molecules are bound to one another based on a modification in the one or more electrical signals to recognize a translocation time for the translocation event in one of the nanochannels;
in response to the large tracer ligand being bound to the investigative molecules while translocating the investigative molecules through the nanochannels, determining that the large tracer ligand remains bound to the investigative molecules due to a long time duration for the large tracer ligand bound to the investigative molecules traveling through the nanochannels, thereby recognizing that the target ligands did not replace the large tracer ligand;
in response to the large tracer ligand being displaced from being bound to the investigative molecules and in response to the large tracer ligand being replaced with the target ligands while translocating the investigative molecules through the nanochannels, determining that the target ligands are bound to the investigative molecules due to a short time duration for the target ligands bound to the investigative molecules traveling through the nanochannels, thereby recognizing that the target ligands have replaced the large tracer ligand;
wherein the short time duration of travel time in the nanochannels is shorter than the long time duration.

6 more elements

Dependencies are inferred from the claim text.

Facts

Application: 13/727,076Filed 26 Dec 2012
Published: US 2014/0179540 A126 Jun 2014
Granted: 17 Oct 2017
Term ends: Not estimatedThe source records don’t support an estimate.

Classification5 codes

G01N 33/6872
Involving proteins, peptides or amino acids: intracellular protein regulatory factors and their receptors, e.g. including ion channels
C07K 14/723
For hormones: g protein coupled receptor, e.g. TSHR-thyrotropin-receptor, LH/hCG receptor, FSH receptor
G01N 2333/726
For hormones: g protein coupled receptor, e.g. TSHR-thyrotropin-receptor, LH/hCG receptor, FSH
G01N
Investigating or analysing materials by determining their chemical or physical properties