WO1997025616A1 - Groupage sanguin par capteur a canal ionique - Google Patents

Groupage sanguin par capteur a canal ionique Download PDF

Info

Publication number
WO1997025616A1
WO1997025616A1 PCT/AU1997/000014 AU9700014W WO9725616A1 WO 1997025616 A1 WO1997025616 A1 WO 1997025616A1 AU 9700014 W AU9700014 W AU 9700014W WO 9725616 A1 WO9725616 A1 WO 9725616A1
Authority
WO
WIPO (PCT)
Prior art keywords
membrane
antibodies
biosensor
molecules
blood
Prior art date
Application number
PCT/AU1997/000014
Other languages
English (en)
Inventor
Vijoleta Lucija Bronislava Braach-Maksvytis
Bruce Andrew Cornell
Original Assignee
Australian Membrane And Biotechnology Research Institute
The University Of Sydney
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 Australian Membrane And Biotechnology Research Institute, The University Of Sydney filed Critical Australian Membrane And Biotechnology Research Institute
Priority to AU13602/97A priority Critical patent/AU1360297A/en
Publication of WO1997025616A1 publication Critical patent/WO1997025616A1/fr

Links

Classifications

    • 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
    • G01N33/5438—Electrodes

Definitions

  • This invention relates to serologic methods and apparatus comprising a membrane biosensor for determining blood type and for identifying antibodies to epitopic sites on blood molecules or large antigens. Further, it relates to the use of a membrane biosensor to determine the immunoglobin class of an antibody.
  • Procedures have also been developed to detect the presence of antibodies to multiple epitopes on a large antigen such as a viral particle, a protein fragment from a cell or a virus or an intact cell.
  • a related procedure has been developed to detect the presence of Abs to a variety of epitope sites on large protein Ags.
  • the prior art method employs deposition of the virus protein fragment or cell (the antigen) onto the surface of an EUSA cell. The patient serum containing the antibodies to a range of eptiopic sites is added.
  • the well volume is washed of free antibody and a reporter anti-Fc antibody tagged with an enzyme label is added and binds to the sample Abs which remain attached to the large Ag, by which means the presence of the bound Ab on the Ag is detected.
  • Biosensors have been constructed comprising biomembranes which are a double layer of closely packed amphiphilic lipid molecules.
  • the molecules of these bilayers exhibit the random motions characteristic of the liquid phase, in which the hydrogen tails of the lipid molecules have sufficient mobility to provide a soft, flexible, viscid surface.
  • the molecules can also diffuse sideways freely within their own monolayer so that two neighbouring lipids in the same monolayer exchange places with each other about once every microsecond, while the lipid molecules in opposite monolayers exchange places on the average of one a year.
  • Ionophores a class of molecules, called ionophores, which facilitate the transport of ions across these membranes.
  • Ion channels are a particular form of ionophore, which as the term implies, are channels through which ions may pass through membranes.
  • a favoured ionophore is gramicidin A which forms aqueous channels in the membrane. Examples of such biosensors are disclosed in the following Internationa Patent Applications, the disclosures of which are incorporated herein by cross reference:
  • the first of these references discloses receptor molecules conjugated with a support that is remote from the receptor site.
  • the support may be a lipid head group, a hydrocarbon chain, a cross-linkable molecule or a membrane protein.
  • the inner level of the membrane may be adjacent a solid surface with groups reactive with the solid surface, and spaced from the surface to provide a reservoir region as disclosed in U.S. Patent No. 5,401,378.
  • Biosensors based on ion channels or ionophores contained within lipid membranes tethered to or deposited onto metal electrodes are disclosed in Australian Patent 623,747 and U.S. patent 5,234,566. Those references disclose a membrane bilayer in which each layer has incorporated therein ionophores and in which the conductance of the membrane is dependent upon the presence of absence of an analyte.
  • the disclosure of Australian Patent 623.747 (incorporated herein by reference) describes various ionophore gating mechanisms termed local disruption gating, extended disruption gating, vertical disruption gating, and extended displacement gating mechanisms to modify the conductivity of the membrane in response to the presence of an analyte.
  • an inner layer of the membrane (the layer closer to the solid electrode surface, if any) contains immobilised or tethered half membrane spanning ion channels which an outer layer contains more mobile half membrane spanning ion channels.
  • One method for immobilising the ion channels of the inner layer is to employ a polymerisable lipid layer and then cross-link the molecules of the inner monolayer and the ionophore. The conductivity of the membrane is altered by the extent to which opposing half membrane spanning ion channels align to establish a membrane spanning channel for ion transmission across the membrane.
  • local disruption gating receptor molecules are linked to mobile ionophores is the outer layer that are aligned with tethered or immobilised ionophores in the inner layer.
  • the introduction of an analyte particle that binds to two adjacent receptors in the outer layer causes the disruption of the orderly alignment of the membrane spanning ionophore.
  • a loss of conductivity occurs due to the deformation of the ionophores of the outer layer caused by the bonding of the analyte with the adjacent receptors.
  • the mechanism of extended disruption gating is similar, except that the displacement of the mobile ionophore is greater.
  • extended disruption gating the binding of pairs of receptors to the same analyte particle cause the outer layer ionophores to move completely out of alignment with the inner layer ionophores.
  • the mechanism of vertical disruption gating is also similar. In that case the presence of the analyte particle bound to two receptor molecules causes a separation of the two layers that disrupts the continuity of the ion channel across the membrane.
  • the mechanism of extended displacement gating utilises two different receptors that bind to each other and are linked receptively to a half membrane ionophore and a membrane molecule.
  • the binding of these two receptor molecules to each other displaces the ionophore and disrupts conductivity.
  • the analyte competes with the second receptor for the binding site on the first receptor.
  • the presence of the analyte breaks the bond between the two receptors and allows the half membrane ionophores to realign and provide an ion conductive path.
  • Each of these mechanisms has in common that the binding of the analyte to the receptor molecule causes a change in the relationship between two half membrane spanning monomers such that the flow of ions across the membrane via the ionophores is allowed or prevented.
  • the present invention employs in one preferred embodiment an ion channel sensor to determine the compatibility of blood products from two samples. It has the advantage of simplifying and speeding the determination of compatibility of the samples.
  • red blood cells from one sample together with plasma from a second sample comprise an analyte that is introduced into a specialised biosensor cell.
  • the present invention consists in a biosensor for detecting the coagulation of blood in an analyte comprising a container for a blood sample subject to testing, said container having a volume for holding a sample of blood products subject to testing, a first electrode in contact with said sample, a second electrode below said volume, means responsive to the change of conduction between said electrodes, a membrane below said volume, said membrane located between said electrodes, and having ion channels therein covalently attached to detector molecules at an upper surface that bind to epitopic sites on blood components, said ion channels having a conductivity to ions that changes upon the binding of said molecules, said volume contacting a filter layer between said membrane and said volume, wherein said blood components if uncoagulated pass said filter and contact said membrane.
  • the membrane further comprises membrane spanning components attached to a second species of molecules that bind to sites on blood components, wherein the presence of a blood component at the surface of the membrane crosslinks the ion channels to said second species of molecules. It is further preferred that the second species of molecules is the same as said detector molecules.
  • the detector molecules and second species of molecules preferably bind to epitopic sites on a blood component, and are preferably antibodies to glycophorin.
  • membrane spanning components are tethered by linking and spacing molecules to said second electrode.
  • the change in conductivity upon the binding to the membrane may be as a result of any gating mechanism such as local disruption gating, extended disruption gating, vertical disruption gating or extended displacement gating.
  • Examples of material which can be used in the filter layer are wellknown to persons skilled in the field and include the material used "BioVue” TM sold by Ortho Diagnostic Systems Inc.
  • the present invention consists in a method of determining compatibility of two blood samples, the method comprising the steps of:
  • the present invention consists in a method of screening for the presence of antibodies or antigens of interest in a biological sample, the method comprising the steps of: (1) introducing the biological sample into a biosensor, the biosensor comprising: a container for biological sample subject to screening, said container having a first electrode in contact with said sample, a second electrode below said sample, means responsive to the change of conduction between said electrodes, a membrane below said sample, said membrane located between said electrodes, and having ion channels therein attached to first detector moieties at an upper surface to which the antibodies or antigens of interest specifically bind, the binding of the antibodies or antigens to the first detector moieties causing a change in the ability of ions to pass through the membrane via the ion channels; and;
  • the membrane further comprises membrane spanning components attached to second detector moieties to which the antibodies or antigens of interest specifically bind, wherein the binding of the antibodies or antigens of interest to the first and second detector moieties crosslinks the ion channels to the membrane spanning components.
  • first and second detector moieties are the same.
  • antibodies of interest in the biological sample that are to be detected and in other embodiments these are antibodies directed against red blood cell antigens or antigens from a pathogen preferrably from a pathogen selected from the group consisting of hepatitis and HTV.
  • antigens of interest in the biological sample are detected.
  • antigens are preferably red blood cell antigens or antigens from a pathogen preferably from a pathogen selected from the group consisting of hepatitis and HIV.
  • the first and second detector moieties are red blood cells or fragments thereof carrying predetermined antigens.
  • the present invention consists in a method of screening for the presence of antibodies in a biological sample directed against distinct epitopic sites on the same antigen, the method comprising the steps of:
  • the biosensor comprising: a container for biological sample subject to screening, said container having a first electrode in contact with said sample, a second electrode below said sample, means responsive to the change of conduction between said electrodes, a membrane below said sample, said membrane located between said electrodes, and having ion channels therein attached to first moieties at an upper surface which bind antibodies; and;
  • the membrane further comprises membrane spanning components attached to second moieties which bind antibodies.
  • first and second detector moieties are the same and are preferably anti-Fc antibodies or Fc binding fragments thereof.
  • further detector moieties attached directly to membrane components other than the membrane spanning components.
  • a range of differing detector moieties attached to the ion channels and/or the membrane spanning components This may be of benefit where it is desired that a range of antibodies or antigens are detected.
  • the cell comprises an ion channel sensor in which the ionophore and membrane spanning lipids (MSL's) present molecules capable of binding to epitopic sites on a red blood cells such that the arrival of the blood cell at the membrane crosslinks the ionophore and MSL and affects the ionic current through the membrane or implements one of the other mechanisms for affecting ionic current through the membrane in a measurable manner.
  • MSL's membrane spanning lipids
  • the mixture of blood products is introduced into the container above a thin layer of preferable glass or plastic percolation beads through which the mixture of samples must pass to reach the membrane.
  • the percolation layer unlike the prior art is capable of having a thickness substantially less than several centimetres.
  • the speed of the process is accomplished by the rapid diffusion of the blood sample though the thin percolation bead layer when no coagulation occurs.
  • the discrimination of the device is accomplished by the different abilities coagulated and uncoagulated blood cells to pass through the filter layer to the membrane.
  • the preferred molecules capable of binding to epitopic sites on a red blood cell are antibodies that selectively bind to glycophorin molecules that present on the surface of the red blood cell.
  • the presence of antibodies to multiple epitopes on a large antigen such as a viral particle, a protein fragment from a cell or a virus or an intact cell is detected by anti-Fc Fab' being attached to both the ion channel and the MSL (membrane spanning lipid) within the sensor membrane.
  • the unbound Ab having been selectively scavenged by an excess of anti-Fc antibodies distributed over the electrode surface.
  • the Abs bound to the large Ag is separated from the unbound free
  • Abs via a bead through which the unbound Abs can pass more readily than the Ab-Ag complex.
  • the percolation of the free Ab to the electrodes may be separated from the percolation of the large Ag-Ab complex.
  • the presence of Abs to a variety of epitopic sites on large protein Ags is detected by tethering the large Ag or Ag fragment to both the MSL and to the mobile outer layer ion channels, such that with the addition of patient serum Ab's to be detected, Abs reactive with the epitopic sites on the Ags cause the Ags to cross link preventing the diffusion of the outer membrane layer of the gA, reducing its admittance, thereby reporting on the binding.
  • a further embodiment of the present invention concerns the determination of the species of an Ab, for example as to whether it's IgG or IgM.
  • the Abs are required to pass through a bead bed labelled with either IgG or IgM molecules preventing the passage of bound antibody to the electrode surface and to the reporter antigens.
  • the invention further provides for the determination of the coagulation state of a population of erythrocytes.
  • the present invention used an array of small diameter (typically 1-100 ⁇ , preferably 10-90 ⁇ ) gold electrodes bearing a biosensor membrane in which the Fab' recognition element was typically anti-glycophorin. Each electrode is independently addressable using a multiplexed array.
  • the addition of a sample of uncoagulated erythrocytes results in a capture and measurement of a binding event at a distribution of the small electrodes reflecting the random dispersal of the erythrocytes in the sample.
  • the addition of a sample of coagulated erythrocytes would result in the measurement of binding at electrodes that were non-randomly distributed over the array.
  • a further embodiment of this approach would be to employ the array to determine cells of differing size and surface antigens.
  • the utility in the latter case being the ability to sort the population of different cell species within the sample.
  • a further embodiment is to use these arrays to may antigenic sites over the surface of individual cells.
  • Figure 1 is schematic representation of the preferred embodiment of the ICS of the present invention.
  • Figure 2 is a schematic representation of the elements of the ICS used to detect the presence of antibodies to multiple epitopes on a large antigen.
  • Figure 3 is a schematic representation of the elements of the ICS used to detect the presence of Abs to a variety of epitopic sites on large protein Ags.
  • Figure 4 is a schematic representation of the array of gold electrodes in a multiplexed array.
  • a biosensor cell 1 for detecting the coagulation of blood in an analyte.
  • the cell is has a volume 3_ for holding a sample of blood products 5 subject to testing.
  • a first electrode 7 is in contact with the sample.
  • a second electrode 9, is below the volume 3_, and below a membrane VI.
  • the electrodes are connected to a voltmeter or impedance measuring device 13 for responding to the change of conduction between said electrodes.
  • the membrane 11 is preferably a amphiphilic bilayer membrane. It comprises membrane spanning lipids 15, half membrane spanning lipids 17 and ionophores 19 below the volume 3_.
  • the ionophores are preferably dimers such as gramicidin A which comprises two monomers that organise into the corresponding layers of the bilayer membrane.
  • the membrane is located between the electrodes so that its electrical impedance properties can be measured by the electrodes.
  • the membrane has its ion channels 29 covalently attached to detector molecules 2_1 at an upper surface that bind to epitopic sites 23 on blood components.
  • the ion channels have a conductivity to ions that changes upon the binding of the molecules 23_ to the blood components.
  • a key component of the invention is a filter layer between the membrane ⁇ and the volume 3. The blood components if uncoagulated will pass through the filter and contact the membrane.
  • the membrane preferably also comprises membrane spanning lipids 25 attached to the same type molecules ⁇ that bind to sites on blood components or to a different type.
  • the presence of a blood component at the surface of the membrane crosslinks the upper portion of an ion channels to a membrane spanning species of molecules.
  • the molecules that binds to the blood cells are an antibody to glycoprotein, a fragment of a derivative thereof.
  • the membrane spanning components 25 may be tethered by linking and spacing molecules to the second electrode 9.
  • the linking and spacing molecules may be those known in the prior art to provide an ionic reservoir region.
  • antibodies to multiple epitopes on a large antigen such as a viral particle, a protein fragment from a cell or a virus or an intact cell are depicted.
  • the sample is introduced as one side of the ICS and allowed to percolate laterally.
  • the presence of one antibody is detected by anti-Fc Fab' being attached to both the ion channel and the MSL (membrane spanning lipid) within the sensor membrane.
  • the unbound Ab is scavenged by an excess of anti-Fc antibodies extending along the electrode surface.
  • the Abs bound to the large Ag are spatially separated from the unbound free Abs.
  • the unbound Abs pass through the bead bed more readily than the Ab-Ag complex. In this way the bound Abs reach a portion of the membrane uncluttered by the unbound Abs. Since the unbound Abs do not cross link the ionophores of the membrane, the change in ion current though the membrane in a cell such as that shown in Figure 1 is due entirely to the presence of the bound Ab.
  • tests may be made for the presence of the antibody to Hepatitis A virus, Hepatitis B core viral proteins, Hepatitis C virus, HTV virus types 1 and 2, HTLV types 1 and 2, p24 antigen, and Hepatitis B surface antigen.
  • Figure 3 depicts the procedures for detecting the presence of Abs to epitopic sites on large protein Ags. This method is appropriate whenever the Ag fragments are characterised by a unique structure.
  • Figure 4 is a perspective view of an array of cells for determining the coagulation states of a population of erythrocytes.
  • the individual cells are constructed as in Figure 1, except that their upper and lower electrodes are connected respectively to the side and bottom edges of the array where they can be polled to determine the conductivity of the individual cells using standard multiplexing electronics. In this manner, the presence of binding on a scale of a tens of microns is achieved. DETECTION OF ANTI-D BLOOD TYPE ANTIGEN
  • Electrodes and electrodes were prepared as described in PCT/AU95/00763. Briefly, electrodes with freshly evaporated gold (lOOoA) on chrome adhesion layer (20 ⁇ A on glass microscope slides) were dipped into an ethanolic solution of the above components, rinsed with ethanol, then stored at 4°C under ethanol until used for impedance measurements. The slide was clamped into a block containing teflon coated wells which defined the area of the working electrode as approximately 16mm z .
  • the 2nd layer was added from an ethanolic solution then phosphate buffered saline (PBS) was added, and the electrode washed 4 times.
  • Streptavidin was added to each well (5 ⁇ l O.lmg/ml in PBS) and incubated 5 minutes before washing out excess unbound streptavidin with PBS.
  • Fab' fragments biotinylated at the free thiol group (Fab' were prepared from IgG antibodies raised against human anti-D blood type antigen) were then added (5 ⁇ l 0.05mg/ml in PBS) and rinsed with PBS after 10 minutes.
  • Reagent red blood cells (3% cell suspension from the "Surgiscreen” kit by Ortho Diagnostic Systems Inc) with and without the anti-D blood type antigen, were added to different electrodes. A change in impedance was observed only in the presence of red cells which possessed the anti-D blood type antigen.

Abstract

Un capteur à canal ionique détermine la compatibilité des produits sanguins à partir de deux échantillons. On introduit les produits sanguins dans une cellule à bicapteur qui comprend un ICS dans lequel l'ionophore et les lipides enjambant la membrane (MSL) présentent des molécules capables de se fixer sur des sites épitopes d'un globule rouge, de façon que l'arrivée de celui-ci au niveau de la membrane réticule l'ionophore et les MSL et affecte le courant ionique traversant la membrane. Les produits sanguins doivent franchir une fine couche de billes de percolation pour atteindre la membrane. Les molécules préférées capables de se fixer sur des sites épitopes d'un globule rouge sont des anticorps qui fixent sélectivement les molécules de glycoprotéines présentes à la furface du globule rouge. Dans un autre mode de réalisation, on détecte la présence d'anticorps de multiples épitopes sur un grand antigène grâce aux fragments Fab' fixés à la fois sur le canal ionique et le MSL à l'intérieur de la membrane du capteur. On détecte la présence d'anticorps d'une variété de sites épitopes sur de grands antigènes protéiques en amarrant le grand antigène ou fragment d'antigène à la fois au MSL et aux canaux ioniques mobiles de la couche externe. On détermine la classe d'un antigène, par exemple s'il s'agit d'une IgG ou d'une IgM, en faisant passer les antigènes par un lit de billes marquées soit par des IgG soit par des IgM empêchant le passage de l'anticorps lié vers la surface de l'électrode et les antigènes reporters. L'invention concerne en outre la détermination de l'état de coagulation d'une population d'érythrocytes à l'aide d'une rangée d'électrodes d'or de petit diamètre (typiquement 1-100 ν, de préférence 10-90 ν) portant une membrane de biocapteur dans laquelle l'élément de reconnaissance des Fab' est typiquement une anti-glycoprotéine. Chaque électrode est indépendamment adressable à l'aide d'un dispositif multiplexé.
PCT/AU1997/000014 1996-01-11 1997-01-13 Groupage sanguin par capteur a canal ionique WO1997025616A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU13602/97A AU1360297A (en) 1996-01-11 1997-01-13 Ion channel sensor typing

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US989996P 1996-01-11 1996-01-11
US60/009,899 1996-01-11

Publications (1)

Publication Number Publication Date
WO1997025616A1 true WO1997025616A1 (fr) 1997-07-17

Family

ID=21740368

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/AU1997/000014 WO1997025616A1 (fr) 1996-01-11 1997-01-13 Groupage sanguin par capteur a canal ionique

Country Status (2)

Country Link
AU (1) AU1360297A (fr)
WO (1) WO1997025616A1 (fr)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999066329A1 (fr) * 1998-06-12 1999-12-23 Cenes Limited Dispositif de criblage a grande capacite
WO2002085926A2 (fr) * 2001-04-19 2002-10-31 GESELLSCHAFT FüR BIOTECHNOLOGISCHE FORSCHUNG MBH (GBF) Procede de production de series d'anticorps stables regenerables
US7384733B1 (en) 1998-12-05 2008-06-10 Xention Discovery Limited Interface patch clamping
US8242077B2 (en) 2001-01-18 2012-08-14 Newcastle University Ventures Limited Biosensor

Citations (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1987000168A2 (fr) * 1985-06-27 1987-01-15 Willis John P Ionophores et membranes selectives aux ions contenant lesdits ionophores
US4713347A (en) * 1985-01-14 1987-12-15 Sensor Diagnostics, Inc. Measurement of ligand/anti-ligand interactions using bulk conductance
GB2195450A (en) * 1986-09-22 1988-04-07 Atomic Energy Authority Uk Sensor
EP0342382A2 (fr) * 1988-05-20 1989-11-23 General Electric Company Détection électrique de la réaction immunologique
AU4078789A (en) * 1988-08-18 1990-03-23 Ambri Limited Improvements in sensitivity and selectivity of ion channel membrane biosensors
AU5033490A (en) * 1989-01-27 1990-08-24 Ambri Limited Receptor membranes and ionophore gating
AU6924591A (en) * 1990-01-09 1991-07-11 Yeda Research And Development Co. Ltd. Biosensors
AU5144493A (en) * 1992-10-01 1994-04-26 Ambri Limited Improved sensor membranes
AU5618894A (en) * 1992-12-03 1994-06-22 Ambri Limited Analyte detection by competitive inhibition of ion channel gating
US5368712A (en) * 1989-11-02 1994-11-29 Synporin Technologies, Inc. Biologically mimetic synthetic ion channel transducers
WO1995008637A1 (fr) * 1993-09-21 1995-03-30 Washington State University Research Foundation Dosage immunologique comportant un recepteur de canal d'ions conjugue a un ligand immobilise dans un film de lipides
WO1995016206A1 (fr) * 1993-12-06 1995-06-15 Biosystems Technology Corporation Composition et procede servant a detecter des analytes
WO1996012957A1 (fr) * 1994-10-19 1996-05-02 Fritz Pittner Biodetecteur de modulation de flux d'ions, regule par bioreconnaissance
AU3864395A (en) * 1994-11-16 1996-06-06 Ambri Limited Detection device and method
AU5640396A (en) * 1995-05-17 1996-11-29 Ambri Limited Improvement in ionic reservoir through application of an electrical potential

Patent Citations (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4713347A (en) * 1985-01-14 1987-12-15 Sensor Diagnostics, Inc. Measurement of ligand/anti-ligand interactions using bulk conductance
WO1987000168A2 (fr) * 1985-06-27 1987-01-15 Willis John P Ionophores et membranes selectives aux ions contenant lesdits ionophores
GB2195450A (en) * 1986-09-22 1988-04-07 Atomic Energy Authority Uk Sensor
EP0342382A2 (fr) * 1988-05-20 1989-11-23 General Electric Company Détection électrique de la réaction immunologique
AU4078789A (en) * 1988-08-18 1990-03-23 Ambri Limited Improvements in sensitivity and selectivity of ion channel membrane biosensors
AU5033490A (en) * 1989-01-27 1990-08-24 Ambri Limited Receptor membranes and ionophore gating
US5368712A (en) * 1989-11-02 1994-11-29 Synporin Technologies, Inc. Biologically mimetic synthetic ion channel transducers
AU6924591A (en) * 1990-01-09 1991-07-11 Yeda Research And Development Co. Ltd. Biosensors
AU5144493A (en) * 1992-10-01 1994-04-26 Ambri Limited Improved sensor membranes
AU5618894A (en) * 1992-12-03 1994-06-22 Ambri Limited Analyte detection by competitive inhibition of ion channel gating
WO1995008637A1 (fr) * 1993-09-21 1995-03-30 Washington State University Research Foundation Dosage immunologique comportant un recepteur de canal d'ions conjugue a un ligand immobilise dans un film de lipides
WO1995016206A1 (fr) * 1993-12-06 1995-06-15 Biosystems Technology Corporation Composition et procede servant a detecter des analytes
WO1996012957A1 (fr) * 1994-10-19 1996-05-02 Fritz Pittner Biodetecteur de modulation de flux d'ions, regule par bioreconnaissance
AU3864395A (en) * 1994-11-16 1996-06-06 Ambri Limited Detection device and method
AU5640396A (en) * 1995-05-17 1996-11-29 Ambri Limited Improvement in ionic reservoir through application of an electrical potential

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999066329A1 (fr) * 1998-06-12 1999-12-23 Cenes Limited Dispositif de criblage a grande capacite
US6936462B1 (en) 1998-06-12 2005-08-30 Xention Discovery Limited High throughput screen
US10006902B2 (en) 1998-06-12 2018-06-26 Sophion Bioscience A/S High throughput screen
US7384733B1 (en) 1998-12-05 2008-06-10 Xention Discovery Limited Interface patch clamping
US8242077B2 (en) 2001-01-18 2012-08-14 Newcastle University Ventures Limited Biosensor
WO2002085926A2 (fr) * 2001-04-19 2002-10-31 GESELLSCHAFT FüR BIOTECHNOLOGISCHE FORSCHUNG MBH (GBF) Procede de production de series d'anticorps stables regenerables
WO2002085926A3 (fr) * 2001-04-19 2003-11-06 Biotechnolog Forschung Gmbh Procede de production de series d'anticorps stables regenerables

Also Published As

Publication number Publication date
AU1360297A (en) 1997-08-01

Similar Documents

Publication Publication Date Title
EP0465266B1 (fr) Immunoessai avec un signal visuel complémentaire
CA2493616C (fr) Dispositif de diagnostic rapide, dosage et tampon multifonctionnel
FI92883C (fi) Testimenetelmä ja reagenssikitti sitä varten
US5827749A (en) Ligand assay method
US5416000A (en) Analyte immunoassay in self-contained apparatus
US6663833B1 (en) Integrated assay device and methods of production and use
JP4851507B2 (ja) 二経路免疫アッセイ装置
KR100575390B1 (ko) 혈액 샘플에서 분석물의 존재를 검출하기 위한 크로마토그래피 분석 장치 및 방법
CA2249303C (fr) Dispositif chromatographique de dosage immunologique et procede d'utilisation de la valence des particules pour la quantification
US5474902A (en) Semi-permeable capillary assay device
US20130102063A1 (en) Small molecules and protein analysis devices based on molecular imprinted polymers
EP2245135A2 (fr) Dispositifs d'analyse de petites molécules et de protéines à partir de polymères à empreintes moléculaires
EP0643834A1 (fr) Analyse simultanee de test anti-dopage et de prise d'empreinte d igitale
US6689317B1 (en) Immunoassay apparatus for diagnosis
JP3135067B2 (ja) 免疫診断検査系およびその利用
WO1997025616A1 (fr) Groupage sanguin par capteur a canal ionique
US5232663A (en) Test carrier for analytical determination having a highly effective fixing layer for flow-through bound/free separation
KR20170005008A (ko) 불완전 항체를 이용하여 혈액형 항원를 검출하기 위한 기기 및 방법
JP3713178B2 (ja) 梅毒抗体検出用免疫分析装置
JP2002214236A (ja) 血中抗原検出方法及び装置
NO153988B (no) Fremgangsmaate for bestemmelse av celletype eller forenlighet paa en fast matrise.
JPS607230B2 (ja) 抗原または抗体を検出する方法
FR2781885A1 (fr) Dispositif et procede electrostatiques de detection immunologique fine
CA2052909A1 (fr) Dispositif servant a executer un dosage manuel rapide

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AL AM AT AU AZ BA BB BG BR BY CA CH CN CU CZ DE DK EE ES FI GB GE HU IL IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MD MG MK MN MW MX NO NZ PL PT RO RU SD SE SG SI SK TJ TM TR TT UA UG US UZ VN AM AZ BY KG KZ MD RU TJ TM

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): KE LS MW SD SZ UG AT BE CH DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
121 Ep: the epo has been informed by wipo that ep was designated in this application
REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

NENP Non-entry into the national phase

Ref country code: JP

Ref document number: 97524685

Format of ref document f/p: F

122 Ep: pct application non-entry in european phase