WO1994025862A1 - Substrat de biocacteur concu pour supporter une membrane lipidique bicouche contenant un recepteur - Google Patents

Substrat de biocacteur concu pour supporter une membrane lipidique bicouche contenant un recepteur Download PDF

Info

Publication number
WO1994025862A1
WO1994025862A1 PCT/US1994/004883 US9404883W WO9425862A1 WO 1994025862 A1 WO1994025862 A1 WO 1994025862A1 US 9404883 W US9404883 W US 9404883W WO 9425862 A1 WO9425862 A1 WO 9425862A1
Authority
WO
WIPO (PCT)
Prior art keywords
substrate
receptor
aperture
membrane
polyimide
Prior art date
Application number
PCT/US1994/004883
Other languages
English (en)
Inventor
Numan S. Dogan
Mete Eray
Bernard J. Van Wie
Original Assignee
Washington State University Research Foundation
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Washington State University Research Foundation filed Critical Washington State University Research Foundation
Publication of WO1994025862A1 publication Critical patent/WO1994025862A1/fr

Links

Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366—Apparatus specially adapted for solid-phase testing
    • G01N33/54373—Apparatus specially adapted for solid-phase testing involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/0627—Sensor or part of a sensor is integrated
    • B01L2300/0636—Integrated biosensor, microarrays
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/0627—Sensor or part of a sensor is integrated
    • B01L2300/0645—Electrodes

Definitions

  • the present invention relates to a substrate for a biosensor which incorporates a receptor into a suspended lipid bilayer membrane.
  • biosensor as used herein is intended to broadly cover devices that use a biologically sensitive material or "receptor” which reacts to stimulus and which can be used, for example, in the detection of chemicals or analytes. Since the cell membranes of both plants and animals are composed of lipids in the form of a lipid bilayer and since various proteins that perform vital cell functions span such a bilayer structure, much work has been performed on model membrane structures.
  • bilayer lipid membranes that are not in the form of a liposome, can be divided into suspended BLM's, where the lipid bilayer is positioned between two aqueous solutions, and supported BLM's where the lipid bilayer is placed on a solid support.
  • the present invention relates to the first type of configuration.
  • the suspended BLMs known heretofore were stable up to only about two hours as contrasted with supported BLMs which are stable for up to about seventy-two hours in one study (but which yield differing specific capacitance values from suspended BLMs due to difficulties in measuring the specific capacitance and resistance) .
  • Liposomes require lengthy techniques for preparation and are less uniform and flexible as compared to suspended BLMs.
  • BLMs offer little or no control over the electrochemical properties of one side of the membrane thus not providing a feasible environment for the incorporation of a variety of biological receptors as compared to the suspended BLMs.
  • the incorporation of receptors in the BLM for the detection of an analyte is also a known technique. The following illustrate certain work of this type:
  • U. J. Krull and coworkers in U.S. Patent No. 4,661,235 illustrate supporting a bilayer lipid membrane using a circular aperture in a sheet of TEFLON fluoropolymer dividing two separate chambers in a housing.
  • U.S. Patent No. 4,874,499 to R. L. Smith et al. illustrates electrochemical microsensors and illustrates (in Fig. 3b) a device having an ion selective "polymeric" membrane, which acts as an ion sensor, placed in a funnel- shaped opening above a cavity holding a silver-silver chloride electrode over a field effect transistor (FET) .
  • FET field effect transistor
  • the structure shown in this patent possesses a funnel shaped geometry having a rectangularly shaped aperture. Apertures that are used in bilayer membrane work are circular in shape for proper support of the lipid bilayer.
  • biosensors comprising membrane proteins (e.g., the acetylcholine receptor protein) reconstituted in polymerized lipid bilayers which are supported in a patch clamp-arrangement adjacent internal and external aqueous compartments that are separated by a glass microelectrode tip.
  • membrane proteins e.g., the acetylcholine receptor protein
  • U.S. Patent No. 5,001,048 to R. F. Taylor et al. illustrates an electrical biosensor containing a biological receptor immobilized and stabilized in a protein film. The protein film is sandwiched between a silica substrate, on one side, and a laminate comprising an equipotential barrier, a reference membrane, and a second silica support, on the other side. 6.
  • U.S. Patent No. 5,111,221 to T. L. Fare et al . also illustrates a receptor-based sensor in which a lipid bilayer containing receptor (s) is deposited onto a porous silicon substrate. w
  • the present invention relates to a biosensor substrate, adapted to mount a bilayer lipid membrane which contains at least one receptor for use in the detection of an analyte in solution, which comprises: 10 (a) a body having side walls and a solid floor which define a chamber to hold a first solution;
  • Fig. 1 is a cross-sectional view of a biosensor which can be used for monitoring the concentration of chemicals in 25 a solution;
  • Fig. 2 is a view similar to that in Fig. 1 which uses a polyimide diaphragm for support of the membrane.
  • Figs . 3 and 4 are views showing the aperture in the polyimide diaphragm or septum over the silicon substrate;
  • Figs. 5A - 5E illustrate the microfabrication process for making the polyimide diaphragm or septum;
  • Fig. 6 illustrates the electrical measurement set-up described in the Example;
  • Figs. 7A -7D illustrate alamethicin activity recorded at
  • Figs. 8A - 8D show acetylcholine channel receptor (AChR) activity recorded under +150 mV holding potential.
  • AChR acetylcholine channel receptor
  • a biosensor containing the biosensor substrate disclosed herein can be made specifically sensitive to a wide variety of biological or chemical materials.
  • the biosensor comprises a detector portion and a transducer.
  • the detector recognizes the desired analyte (s) in the adjacent environment using a suitable receptor, embedded in a bilayer lipid membrane, which generates a signal upon detection of the target species.
  • a preferred class of receptor are several ion channel forming proteins.
  • the function of the transducer portion of the biosensor is to couple the detected signal into a suitable storage unit, such as a personal computer.
  • the transducer component of the biosensor consists of the physical unit (or "substrate") carrying the bilayer lipid membrane, the compartment (s) for the bathing solution, and the electrodes to monitor current generated.
  • a biosensor should be highly specific and sensitive towards the target analyte(s), should be durable, miniaturizable and easily interfaced with the selected measurement and data storage units.
  • the specificity is dependent upon the receptor chosen for incorporation in the BLM and the inertness of the lipid material to the target analyte(s) .
  • the sensitivity is dependent upon the receptor protein used as well as the measurement system that is selected. Although either amperometric, potentiometric, or optical measurement modes can be selected, it is preferred to select the amperometric mode which has the highest inherent sensitivity (10' 7 to 10* mole/liter).
  • Figs. 1 and 2 illustrate preferred embodiments of the present biosensor substrate.
  • the biosensor substrate comprises a body 2 which has side walls 9 and a floor 10 which define a chamber 8 adapted to hold an aqueous solution.
  • the body 2 is formed of micromachined silicon with the side walls being formed of glass.
  • a glass shield structure 1 is placed beneath body 2 to enclose a second chamber 3 which is adapted to also hold an aqueous solution.
  • These glass structures 1 and 9 provide access to suitable electrodes 4 and 7 which can be silver-silver chloride electrodes.
  • the bilayer lipid membrane 5 contains reincorporated active membrane proteins, for example, an ion channel forming protein such as a nicotinic acetylcholine receptor 6 and spans an aperture in the floor 10. It preferably is round (or circular) with a diameter of from about 1 ⁇ m to about 50 ⁇ m.
  • Fig. 2 shows an analogous structure in which chambers 13 and 18 for the aqueous solutions are separated by a micromachined silicon body 12, for example, having, as the floor 20, a polyimide which contains the bilayer lipid membrane 15 and incorporated receptor 16. Glass side wall shield 19 and glass shield 11 enclose the respective aqueous chambers or compartments 18 and 13 and provide access to the electrodes 17 and 14.
  • polyimide as a support for the bilipid membrane has several advantages. First, the sealing obtained in a device that uses polyimide as the membrane support is appreciably better as compared to a device using a silicon membrane support as depicted in Fig. 1.
  • polyimide is hydrophobic, silanization, which is required for silicon devices, is not needed thereby eliminating the time consuming and tedious silanization process required for silicon membrane support structures.
  • polyimide is transparent to light which lends to use of optical detection of molecular binding events, for example, by coupling light from a fiber into the polyimide film to excite the bilipid membrane with measurement of the optical response of the membrane to detect analyte-receptor interactions.
  • the biosensor substrate of this invention allows for the fabrication of a compact, solid state biosensor which can detect the presence of environmental stimulus by having electrodes detect voltage and current changes across a reconstituted bilayer lipid membrane containing receptor proteins therein which react to a specific stimulus.
  • the support structure extends the longevity of the lipid bilayer component used in such a biosensor. Since the lipid membrane is formed on the same substrate along with portions of the electronic circuit that monitors and records the electrical activity from the membrane-receptor combination, there is good compatibility between the electronic and biological detector subcomponents.
  • Photosensitive polyimide (Pyralin PI2722 brand) was obtained from DuPont Electronics (Wilmington, DE) .
  • the developing solutions were mixtures of ⁇ -hydroxybutyric acid (obtained from Sigma, St. Louis, MO), and xylenes (from Baker Analyzed, Phillipsburg, NJ) .
  • Black Teflon fluoropolymer for two detachable chambers (for the trans and cis sides of the BLM) was purchased from Laird Plastic (Spokane, WA) . Silicon substrate carrying the polyimide septum was attached to one of the detachable chambers with Teflon epoxy (Duralco 4540 brand obtained from Cotronics Co., Brooklyn, NY).
  • Parts of the electrical measurement set-up namely a CV-201 brand current to voltage (I/V) converter headstage, Axopatch 200 brand patch-clamp amplifier and TL-1 A/D brand converter were obtained from Axon Instruments (Foster City, CA) .
  • Lecithin type II-S, n-decane, and alamethicin were purchased from Sigma (St. Louis, MO) .
  • FIG. 3 A schematical diagram of the designed polyimide septum is shown in Fig. 3.
  • the silica layer between the silicon and polyimide is a sacrificial layer that is used to protect the polyimide from the anisotropic KOH etchant.
  • the aim was to obtain an aperture with diameter, R, equal to 50 ⁇ m, and an aspect ratio of about 0.1.
  • the microlithography steps used in the fabrication process of polyimide septa are outlined in Fig. 4.
  • a contact printing technique was used throughout the process and is described in detail by W. R. Runyan et al., Semiconductor Integrated Circuit Processing Technology
  • n-type ⁇ 100> oriented 3 inch silicon (Si) wafers were agitated in a sulfuric acid»hydrogen peroxide solution for ten minutes. After a through rinse under deionized water (DIW) , they were cleaned of organic contaminants by rinsing under trichloroethylene, acetone, methanol, and finally under DIW. Next, 1 ⁇ m thick silica layers were thermally grown on both sides of the wafers. Polyimide stock solution was thawed to room temperature before each process run. Polyimide was spun on the front surface of a silicon wafer for forty seconds, at 4000 rpm, forming a uniform layer.
  • DIW deionized water
  • the polyimide layer was exposed for one minute using a 450 nm UV light source.
  • Four developer solutions with volume ratios 5:5, 6:4, 7:3, 8:2 of xylenes: ⁇ - hydroxybutyric acid were prepared and sprayed on the exposed surface with durations of twenty, sixteen, twelve and eight seconds, respectively.
  • the electrical measurement set-up is shown in Fig. 5 and is very similar to the design given by Alvarez in Ion Channel Reconstitution (Plenum Press, New York, 1986) pp. 115-130.
  • the fluoropolymer cuvette was basically a detachable unit of two pieces.
  • the silicon partition carrying the polyimide septum was fixed on one piece using Duralco 4540 brand Teflon fluoropolymer epoxy.
  • An optional silicon sealant layer was applied to the surface to reduce the force on silicon chips during attachment. After proper curing of both adhesives, the two Teflon fluoropolymer units were snugly fit together using long screws with the polyimide septum in between.
  • the CV-201 brand apparatus is a capacitive headstage for current- voltage (I/V) conversion and the Axopatch 200 apparatus is a patch-clamp amplifier which differentiates the headstage output signal, filters it and finally amplifies the signal.
  • the waveform generator gives a 125 Hz, 20 mV p _ p triangular waveform that was used in the specific capacitance and resistance measurements.
  • the TL-1 A/D brand converter provided digitized forms of Axopatch 200 measurements to a IBM PC-compatible computer.
  • Lipid solutions Alamethicin channels, and AChR vesicles
  • the standard lipid solution was 10 mg of lecithin type
  • the polyimide septum supported on the silicon rim contained an aperture with a diameter of about 40 ⁇ m and a thickness of about 6 ⁇ m. Thus, an aspect ratio of about 0.1 was obtained which is suitable for stirring purposes.
  • the extremely smooth tapered aperture edges and septum surface were evident in micrographs taken of the structures, were formed as a result of the UV microlithography procedure and are important features for long term stability of the lipid bilayer.
  • the polyimide septum was mechanically stable and did not require any preconditioning before the application of the lipid solutions.
  • the adhesion of the polyimide to the sacrificial silica layer was excellent.
  • the polyimide septum was completely dried at about 80°C in an oven before BLM formation. Then 7 ⁇ l of lipid solution was gently dropped over the aperture using a micropipette holder. Within the next twenty seconds, after the lipid solution was applied to the aperture, both chambers of the
  • Teflon fluoropolymer cuvette were filled up very slowly with appropriate salt solutions. Over about the next thirty seconds, thinning of the BLM could be observed with an increasing capacitance.
  • Collective capacitance of the polyimide septum and BLM stabilized at 220 pF. After a month of experiments, the aperture was plugged with a small amount of polyimide and cured. Then, the intrinsic capacitance of the septum was measured to be 200 pF. This gives a specific capacitance of 1.59 ⁇ F/cm 2 for the BLM as compared to Tien, in Bilayer Lipid Membranes Theory and Practice (Marcel
  • Alamethicin is a membrane spanning ion channel. If the membrane is not a bilayer, the alamethicin activity cannot be observed (Alvarez et al., supra, 1981).
  • 2 ⁇ l of alamethicin stock solution was applied to the cis side of BLM.
  • channel activity was observed.
  • Part of the data recorded at a +150 mV holding potential (cis side positive) is given in Fig. 6. Three main activity levels with conductivities of 61 pU, 85 pU, and 115 pU, respectively, were observed.
  • AChR stock solution was thawed to room temperature and was very gently sonicated. Both sides of the BLM were composed of 20 mM potassium chloride and 10 mM hepes (pH - 7.4). Then, 2 ⁇ l of stock solution was added to the cis side of the BLM under constant stirring conditions and +150 mV applied potential. Within the next ten minutes, no activity was observed. Next, 5 ⁇ M of calcium chloride was added to the cis side to promote fusion by attracting the vesicles to the vicinity of BLM, a state called perfusion by Cohen et al. in J. Cell. Biol. 98, 1054- 1062 (1984) . Still, no activity was observed within the next ten minutes.
  • FIG. 7 shows the AChR activity. Two main activities with conductances of 61 pU, and 140 pU were observed which are believed to be the sub- and main conductance levels of a single AChR molecule. Similar activity for AChRs in the synaptic membrane of skeletal muscle was observed by Hamill et al. in Nature 294, 462-464 (1981) with conductance levels of 25 and 35 p ⁇ s.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Hematology (AREA)
  • Biomedical Technology (AREA)
  • General Health & Medical Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Molecular Biology (AREA)
  • Urology & Nephrology (AREA)
  • Food Science & Technology (AREA)
  • Microbiology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Cell Biology (AREA)
  • Biochemistry (AREA)
  • Biotechnology (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Dispersion Chemistry (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

Un substrat de biocacteur comprend: (a) un corps (2) possédant des parois latérales (9) et une base solide (10) formant une chambre (8) destinée à une première solution; (b) au moins une ouverture ouverte se trouvant dans la base solide pour permettre l'intégrité structurale d'une membrane lipidique bicouche (5) contenant un récepteur d'analyte (par exemple, un récepteur de canal ionique) (6) dans l'ouverture; et (c) une chambre (3) située sous la base solide et destinée à contenir une seconde solution se trouvant en contact avec la membrane contenant le récepteur.
PCT/US1994/004883 1993-05-04 1994-05-03 Substrat de biocacteur concu pour supporter une membrane lipidique bicouche contenant un recepteur WO1994025862A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US5727593A 1993-05-04 1993-05-04
US08/057,275 1993-05-04

Publications (1)

Publication Number Publication Date
WO1994025862A1 true WO1994025862A1 (fr) 1994-11-10

Family

ID=22009596

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1994/004883 WO1994025862A1 (fr) 1993-05-04 1994-05-03 Substrat de biocacteur concu pour supporter une membrane lipidique bicouche contenant un recepteur

Country Status (1)

Country Link
WO (1) WO1994025862A1 (fr)

Cited By (40)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0873789A2 (fr) * 1997-04-21 1998-10-28 Fraunhofer-Gesellschaft Zur Förderung Der Angewandten Forschung E.V. Dispositif avec chambre encapsulant un materiau
WO2000025121A1 (fr) * 1998-10-27 2000-05-04 President And Fellows Of Harvard College Canaux ioniques biologiques dans des detecteurs nanofabriques
WO2000079257A1 (fr) * 1999-06-22 2000-12-28 President And Fellows Of Harvard College Evaluation a l'echelle atomique et moleculaire de biopolymeres
WO2001048475A1 (fr) * 1999-12-24 2001-07-05 Astrazeneca Ab Dispositif et procede permettant de faire des mesures electriques sur un objet
WO2001048474A1 (fr) * 1999-12-24 2001-07-05 Astrazeneca Ab Appareil et procede destines a realiser des mesures electriques sur des objets
US6267872B1 (en) * 1998-11-06 2001-07-31 The Regents Of The University Of California Miniature support for thin films containing single channels or nanopores and methods for using same
WO2001059447A1 (fr) * 2000-02-11 2001-08-16 Yale University Electrodes patch-clamp planes
EP1225216A1 (fr) * 2001-01-08 2002-07-24 Niels Fertig Appareil pour analyser des canaux ioniques dans un membrane
GB2371626A (en) * 2000-10-11 2002-07-31 Axon Instr Inc Positioning cells with high resistance seal
FR2836071A1 (fr) * 2002-02-21 2003-08-22 Commissariat Energie Atomique Composant pour microsysteme d'analyse biologique ou biochimique
US6673615B2 (en) 1995-03-17 2004-01-06 President And Fellows Of Harvard College Characterization of individual polymer molecules based on monomer-interface interactions
US6682649B1 (en) 1999-10-01 2004-01-27 Sophion Bioscience A/S Substrate and a method for determining and/or monitoring electrophysiological properties of ion channels
WO2004035211A1 (fr) * 2002-10-15 2004-04-29 Advanced Research Corporation Dispositif de canal a membrane semi-conductrice pour la mesure et la caracterisation d'echantillons de tailles atomique et moleculaire
WO2005040783A1 (fr) * 2003-10-22 2005-05-06 Ambri Limited Nouvelle configuration de capteur
EP1526110A3 (fr) * 2003-10-24 2005-10-12 Agilent Technologies Inc. (a Delaware Corporation) Chip en semiconducteur de type n aven un nanopore.
EP1712909A1 (fr) * 2004-01-21 2006-10-18 Japan Science and Technology Agency Methode permettant de former une membrane lipidique double plane a des fins d'analyse proteique membranaire
WO2007001410A2 (fr) * 2004-10-14 2007-01-04 Electronic Biosciences, Llc Reseau de capteurs integre a des fins de production d'une bioempreinte d'analyte
US7169272B2 (en) * 1997-04-30 2007-01-30 Board Of Trustees Of The University Of Arkansas Microfabricated recessed disk microelectrodes: characterization in static and convective solutions
US7238485B2 (en) 2004-03-23 2007-07-03 President And Fellows Of Harvard College Methods and apparatus for characterizing polynucleotides
GB2447043A (en) * 2007-02-20 2008-09-03 Oxford Nanolabs Ltd Lipid bilayer sensor system
EP2107040A1 (fr) 2008-03-31 2009-10-07 Sony Deutschland Gmbh Procédé de fabrication d'une membrane disposant d'un pore conique
WO2009077734A3 (fr) * 2007-12-19 2009-10-15 Oxford Nanopore Technologies Limited Formation de couches de molécules amphiphiles
US7723029B2 (en) 2002-01-24 2010-05-25 Aviva Biosciences Corporation Biochips including ion transport detecting structures and methods of use
US7846738B2 (en) 2003-08-15 2010-12-07 President And Fellows Of Harvard College Study of polymer molecules and conformations with a nanopore
US7968305B2 (en) 2001-03-24 2011-06-28 Aviva Biosciences Corporation Biochips including ion transport detecting structures and methods of use
US20110174625A1 (en) * 2007-04-04 2011-07-21 Akeson Mark A Compositions, devices, systems, and methods for using a nanopore
CN102216773A (zh) * 2009-10-07 2011-10-12 松下电器产业株式会社 人工脂质膜形成方法
US8206568B2 (en) * 1999-06-22 2012-06-26 President And Fellows Of Harvard College Material deposition techniques for control of solid state aperture surface properties
US8828211B2 (en) 2010-06-08 2014-09-09 President And Fellows Of Harvard College Nanopore device with graphene supported artificial lipid membrane
US8986528B2 (en) 1995-03-17 2015-03-24 President And Fellows Of Harvard College Characterization of individual polymer molecules based on monomer-interface interactions
US9146221B2 (en) 2001-03-24 2015-09-29 Aviva Biosciences Corporation High-density ion transport measurement biochip devices and methods
US9377437B2 (en) 2010-02-08 2016-06-28 Genia Technologies, Inc. Systems and methods for characterizing a molecule
US10006902B2 (en) 1998-06-12 2018-06-26 Sophion Bioscience A/S High throughput screen
US10215768B2 (en) 2007-02-20 2019-02-26 Oxford Nanopore Technologies Ltd. Lipid bilayer sensor system
US10338056B2 (en) 2012-02-13 2019-07-02 Oxford Nanopore Technologies Ltd. Apparatus for supporting an array of layers of amphiphilic molecules and method of forming an array of layers of amphiphilic molecules
US10359427B2 (en) * 2016-01-08 2019-07-23 Washington State University Dual ionophore ion selective electrode for detecting non-ionic molecules, macromolecules and cells
US10549274B2 (en) 2014-10-17 2020-02-04 Oxford Nanopore Technologies Ltd. Electrical device with detachable components
US10814298B2 (en) 2012-10-26 2020-10-27 Oxford Nanopore Technologies Ltd. Formation of array of membranes and apparatus therefor
US11596940B2 (en) 2016-07-06 2023-03-07 Oxford Nanopore Technologies Plc Microfluidic device
US11789006B2 (en) 2019-03-12 2023-10-17 Oxford Nanopore Technologies Plc Nanopore sensing device, components and method of operation

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3856633A (en) * 1971-01-07 1974-12-24 Foxboro Co Concentration measurements utilizing coulometric generation of reagents
US4456522A (en) * 1981-09-23 1984-06-26 Critikon, Inc. Support and anchoring mechanism for membranes in selectively responsive field effect devices
USH201H (en) * 1985-08-23 1987-01-06 United States Of America Biosensors from membrane proteins reconstituted in polymerized lipid bilayers
US4661235A (en) * 1984-08-03 1987-04-28 Krull Ulrich J Chemo-receptive lipid based membrane transducers
US4874499A (en) * 1988-05-23 1989-10-17 Massachusetts Institute Of Technology Electrochemical microsensors and method of making such sensors
US5001048A (en) * 1987-06-05 1991-03-19 Aurthur D. Little, Inc. Electrical biosensor containing a biological receptor immobilized and stabilized in a protein film
US5111221A (en) * 1988-05-13 1992-05-05 United States Of America As Represented By The Secretary Of The Navy Receptor-based sensor

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3856633A (en) * 1971-01-07 1974-12-24 Foxboro Co Concentration measurements utilizing coulometric generation of reagents
US4456522A (en) * 1981-09-23 1984-06-26 Critikon, Inc. Support and anchoring mechanism for membranes in selectively responsive field effect devices
US4661235A (en) * 1984-08-03 1987-04-28 Krull Ulrich J Chemo-receptive lipid based membrane transducers
USH201H (en) * 1985-08-23 1987-01-06 United States Of America Biosensors from membrane proteins reconstituted in polymerized lipid bilayers
US5001048A (en) * 1987-06-05 1991-03-19 Aurthur D. Little, Inc. Electrical biosensor containing a biological receptor immobilized and stabilized in a protein film
US5111221A (en) * 1988-05-13 1992-05-05 United States Of America As Represented By The Secretary Of The Navy Receptor-based sensor
US4874499A (en) * 1988-05-23 1989-10-17 Massachusetts Institute Of Technology Electrochemical microsensors and method of making such sensors

Cited By (75)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6673615B2 (en) 1995-03-17 2004-01-06 President And Fellows Of Harvard College Characterization of individual polymer molecules based on monomer-interface interactions
US8986528B2 (en) 1995-03-17 2015-03-24 President And Fellows Of Harvard College Characterization of individual polymer molecules based on monomer-interface interactions
US9046483B2 (en) 1995-03-17 2015-06-02 President And Fellows Of Harvard College Characterization of individual polymer molecules based on monomer-interface interactions
US7189503B2 (en) 1995-03-17 2007-03-13 President And Fellows Of Harvard College Characterization of individual polymer molecules based on monomer-interface interactions
EP0873789A3 (fr) * 1997-04-21 1999-09-15 Fraunhofer-Gesellschaft Zur Förderung Der Angewandten Forschung E.V. Dispositif avec chambre encapsulant un materiau
EP0873789A2 (fr) * 1997-04-21 1998-10-28 Fraunhofer-Gesellschaft Zur Förderung Der Angewandten Forschung E.V. Dispositif avec chambre encapsulant un materiau
US7169272B2 (en) * 1997-04-30 2007-01-30 Board Of Trustees Of The University Of Arkansas Microfabricated recessed disk microelectrodes: characterization in static and convective solutions
US10006902B2 (en) 1998-06-12 2018-06-26 Sophion Bioscience A/S High throughput screen
WO2000025121A1 (fr) * 1998-10-27 2000-05-04 President And Fellows Of Harvard College Canaux ioniques biologiques dans des detecteurs nanofabriques
EP1135676A4 (fr) * 1998-11-06 2002-04-03 Univ California Support miniature pour films minces contenant des canaux uniques ou des nanopores et procedes d'utilisation de ces derniers
US6267872B1 (en) * 1998-11-06 2001-07-31 The Regents Of The University Of California Miniature support for thin films containing single channels or nanopores and methods for using same
US6746594B2 (en) 1998-11-06 2004-06-08 The Regents Of The University Of California Miniature support for thin films containing single channels or nanopores and methods for using the same
EP1135676A1 (fr) * 1998-11-06 2001-09-26 The Regents of the University of California Support miniature pour films minces contenant des canaux uniques ou des nanopores et procedes d'utilisation de ces derniers
US6627067B1 (en) 1999-06-22 2003-09-30 President And Fellows Of Harvard College Molecular and atomic scale evaluation of biopolymers
WO2000079257A1 (fr) * 1999-06-22 2000-12-28 President And Fellows Of Harvard College Evaluation a l'echelle atomique et moleculaire de biopolymeres
US8206568B2 (en) * 1999-06-22 2012-06-26 President And Fellows Of Harvard College Material deposition techniques for control of solid state aperture surface properties
US6682649B1 (en) 1999-10-01 2004-01-27 Sophion Bioscience A/S Substrate and a method for determining and/or monitoring electrophysiological properties of ion channels
WO2001048475A1 (fr) * 1999-12-24 2001-07-05 Astrazeneca Ab Dispositif et procede permettant de faire des mesures electriques sur un objet
WO2001048474A1 (fr) * 1999-12-24 2001-07-05 Astrazeneca Ab Appareil et procede destines a realiser des mesures electriques sur des objets
US6699697B2 (en) 2000-02-11 2004-03-02 Yale University Planar patch clamp electrodes
WO2001059447A1 (fr) * 2000-02-11 2001-08-16 Yale University Electrodes patch-clamp planes
GB2371626B (en) * 2000-10-11 2005-03-16 Axon Instr Inc Parallel electrode assembly and method of positioning cells for electrophysiological testing
GB2371626A (en) * 2000-10-11 2002-07-31 Axon Instr Inc Positioning cells with high resistance seal
EP1225216A1 (fr) * 2001-01-08 2002-07-24 Niels Fertig Appareil pour analyser des canaux ioniques dans un membrane
WO2002066596A3 (fr) * 2001-01-08 2003-03-27 Niels Fertig Dispositif et procede d'analyse de canaux ioniques dans des membranes
WO2002066596A2 (fr) * 2001-01-08 2002-08-29 Niels Fertig Dispositif et procede d'analyse de canaux ioniques dans des membranes
US7968305B2 (en) 2001-03-24 2011-06-28 Aviva Biosciences Corporation Biochips including ion transport detecting structures and methods of use
US9146221B2 (en) 2001-03-24 2015-09-29 Aviva Biosciences Corporation High-density ion transport measurement biochip devices and methods
US7723029B2 (en) 2002-01-24 2010-05-25 Aviva Biosciences Corporation Biochips including ion transport detecting structures and methods of use
WO2003071278A1 (fr) * 2002-02-21 2003-08-28 Commissariat A L'energie Atomique Composant pour microsysteme d'analyse biologique ou biochimique
FR2836071A1 (fr) * 2002-02-21 2003-08-22 Commissariat Energie Atomique Composant pour microsysteme d'analyse biologique ou biochimique
WO2004035211A1 (fr) * 2002-10-15 2004-04-29 Advanced Research Corporation Dispositif de canal a membrane semi-conductrice pour la mesure et la caracterisation d'echantillons de tailles atomique et moleculaire
US8969091B2 (en) 2003-08-15 2015-03-03 President And Fellows Of Harvard College Study of polymer molecules and conformations with a nanopore
US8394640B2 (en) 2003-08-15 2013-03-12 President And Fellows Of Harvard College Study of polymer molecules and conformations with a nanopore
US7846738B2 (en) 2003-08-15 2010-12-07 President And Fellows Of Harvard College Study of polymer molecules and conformations with a nanopore
WO2005040783A1 (fr) * 2003-10-22 2005-05-06 Ambri Limited Nouvelle configuration de capteur
EP1526110A3 (fr) * 2003-10-24 2005-10-12 Agilent Technologies Inc. (a Delaware Corporation) Chip en semiconducteur de type n aven un nanopore.
EP1712909A1 (fr) * 2004-01-21 2006-10-18 Japan Science and Technology Agency Methode permettant de former une membrane lipidique double plane a des fins d'analyse proteique membranaire
EP1712909A4 (fr) * 2004-01-21 2008-11-19 Japan Science & Tech Agency Methode permettant de former une membrane lipidique double plane a des fins d'analyse proteique membranaire
US7625706B2 (en) 2004-03-23 2009-12-01 Agilent Technologies, Inc. Methods and apparatus for characterizing polynucleotides
US8673556B2 (en) 2004-03-23 2014-03-18 President And Fellows Of Harvard College Methods and apparatus for characterizing polynucleotides
US7238485B2 (en) 2004-03-23 2007-07-03 President And Fellows Of Harvard College Methods and apparatus for characterizing polynucleotides
US7947454B2 (en) 2004-03-23 2011-05-24 President And Fellows Of Harvard College Methods and apparatus for characterizing polynucleotides
WO2007001410A3 (fr) * 2004-10-14 2007-05-18 Electronic Biosciences Llc Reseau de capteurs integre a des fins de production d'une bioempreinte d'analyte
GB2432424B (en) * 2004-10-14 2008-09-10 Electronic Bio Sciences Llc Integrated sensing array for producing a biofinger print of an analyte
WO2007001410A2 (fr) * 2004-10-14 2007-01-04 Electronic Biosciences, Llc Reseau de capteurs integre a des fins de production d'une bioempreinte d'analyte
US10215768B2 (en) 2007-02-20 2019-02-26 Oxford Nanopore Technologies Ltd. Lipid bilayer sensor system
GB2447043A (en) * 2007-02-20 2008-09-03 Oxford Nanolabs Ltd Lipid bilayer sensor system
US10081835B2 (en) 2007-04-04 2018-09-25 The Regents Of The University Of California Nucleotide sequencing using an array of independently addressable nanopores
US10059988B2 (en) 2007-04-04 2018-08-28 The Regents Of The University Of California Methods for using a nanopore
US8679747B2 (en) 2007-04-04 2014-03-25 The Regents Of The University Of California Compositions, devices, systems, for using a nanopore
US20110174625A1 (en) * 2007-04-04 2011-07-21 Akeson Mark A Compositions, devices, systems, and methods for using a nanopore
US10344327B2 (en) 2007-04-04 2019-07-09 The Regents Of The University Of California Compositions, devices, systems, and methods for using a nanopore
US8500982B2 (en) * 2007-04-04 2013-08-06 The Regents Of The University Of California Compositions, devices, systems, and methods for using a nanopore
US10208342B2 (en) 2007-04-04 2019-02-19 The Regents Of The University Of California Compositions, devices, systems, and methods for using a nanopore
US9797013B2 (en) 2007-04-04 2017-10-24 The Regents Of The University Of California Compositions, devices, systems, and methods for using a nanopore
US10202645B2 (en) 2007-04-04 2019-02-12 The Regents Of The University Of California Compositions, devices, systems, and methods for using a nanopore
US10196688B2 (en) 2007-04-04 2019-02-05 The Regents Of The University Of California Compositions, devices, systems, and methods for using a nanopore
WO2009077734A3 (fr) * 2007-12-19 2009-10-15 Oxford Nanopore Technologies Limited Formation de couches de molécules amphiphiles
US9927398B2 (en) 2007-12-19 2018-03-27 Oxford Nanopore Technologies Ltd. Formation of layers of amphiphilic molecules
US11898984B2 (en) 2007-12-19 2024-02-13 Oxford Nanopore Technologies Plc Nanopore arrays for sequencing nucleic acids
US10416117B2 (en) 2007-12-19 2019-09-17 Oxford Nanopore Technologies Ltd. Formation of layers of amphiphilic molecules
EP2107040A1 (fr) 2008-03-31 2009-10-07 Sony Deutschland Gmbh Procédé de fabrication d'une membrane disposant d'un pore conique
CN102216773A (zh) * 2009-10-07 2011-10-12 松下电器产业株式会社 人工脂质膜形成方法
US9377437B2 (en) 2010-02-08 2016-06-28 Genia Technologies, Inc. Systems and methods for characterizing a molecule
US8828211B2 (en) 2010-06-08 2014-09-09 President And Fellows Of Harvard College Nanopore device with graphene supported artificial lipid membrane
US9797863B2 (en) 2010-06-08 2017-10-24 President And Fellows Of Harvard College Graphene supported artificial membranes and uses thereof
US10338056B2 (en) 2012-02-13 2019-07-02 Oxford Nanopore Technologies Ltd. Apparatus for supporting an array of layers of amphiphilic molecules and method of forming an array of layers of amphiphilic molecules
US11561216B2 (en) 2012-02-13 2023-01-24 Oxford Nanopore Technologies Plc Apparatus for supporting an array of layers of amphiphilic molecules and method of forming an array of layers of amphiphilic molecules
US11913936B2 (en) 2012-02-13 2024-02-27 Oxford Nanopore Technologies Plc Apparatus for supporting an array of layers of amphiphilic molecules and method of forming an array of layers of amphiphilic molecules
US10814298B2 (en) 2012-10-26 2020-10-27 Oxford Nanopore Technologies Ltd. Formation of array of membranes and apparatus therefor
US10549274B2 (en) 2014-10-17 2020-02-04 Oxford Nanopore Technologies Ltd. Electrical device with detachable components
US10359427B2 (en) * 2016-01-08 2019-07-23 Washington State University Dual ionophore ion selective electrode for detecting non-ionic molecules, macromolecules and cells
US11596940B2 (en) 2016-07-06 2023-03-07 Oxford Nanopore Technologies Plc Microfluidic device
US11789006B2 (en) 2019-03-12 2023-10-17 Oxford Nanopore Technologies Plc Nanopore sensing device, components and method of operation

Similar Documents

Publication Publication Date Title
WO1994025862A1 (fr) Substrat de biocacteur concu pour supporter une membrane lipidique bicouche contenant un recepteur
Lambrechts et al. Biosensors: microelectrochemical devices
ES2333831T3 (es) Sensor basado en aberturas microfabricadas.
US6699697B2 (en) Planar patch clamp electrodes
Nikolelis et al. Ammonium ion minisensors from self-assembled bilayer lipid membranes using gramicidin as an ionophore. Modulation of ammonium selectivity by platelet-activating factor
Mădăraş et al. Microfabricated amperometric creatine and creatinine biosensors
US5001048A (en) Electrical biosensor containing a biological receptor immobilized and stabilized in a protein film
US5352348A (en) Method of using enzyme electrode
US6863833B1 (en) Microfabricated apertures for supporting bilayer lipid membranes
EP1322955B1 (fr) Systeme permettant de proceder a des mesures electrophysiologiques
Cheng et al. Single ion channel sensitivity in suspended bilayers on micromachined supports
Nikolelis et al. Bilayer lipid membranes for electrochemical sensing
EP0080601A1 (fr) Membrane pour électrode enzymatique, procédé de fabrication d'une telle membrane et structure de cellule polarographique
US6852525B1 (en) Mammalian cell culture chamber
JPH0418626B2 (fr)
CA2348002A1 (fr) Canaux ioniques biologiques dans des detecteurs nanofabriques
Nikolelis et al. Stabilized lipid film based biosensor for atenolol
Yang et al. Biosensors and their Applications
Campanella et al. Organophosphorus pesticide (Paraoxon) analysis using solid state sensors
Eray et al. Highly stable bilayer lipid membranes (BLMs) formed on microfabricated polyimide apertures
US20090311670A1 (en) Apparatus and method for measuring concentration of molecules through a barrier
Higson et al. Biosensors: a viable monitoring technology?
Tien Bilayer lipid membrane‐based electrochemical biosensors
US20040168912A1 (en) Planar patch clamp electrodes
JP3665720B2 (ja) センサデバイス

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): CA JP

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FR GB GR IE IT LU MC NL PT SE

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
122 Ep: pct application non-entry in european phase
NENP Non-entry into the national phase

Ref country code: CA