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WO1999044046A1 - Cartouche equipee d'un detecteur de proteine et dispositif de lecture associe - Google Patents

Cartouche equipee d'un detecteur de proteine et dispositif de lecture associe Download PDF

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Publication number
WO1999044046A1
WO1999044046A1 PCT/NL1999/000085 NL9900085W WO9944046A1 WO 1999044046 A1 WO1999044046 A1 WO 1999044046A1 NL 9900085 W NL9900085 W NL 9900085W WO 9944046 A1 WO9944046 A1 WO 9944046A1
Authority
WO
WIPO (PCT)
Prior art keywords
contact
sensor
analyte
reagent
sensor device
Prior art date
Legal status (The legal status 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 status listed.)
Ceased
Application number
PCT/NL1999/000085
Other languages
English (en)
Inventor
Piet Bergveld
Gerardus Antonius Johannes Besselink
Sebastian BÖHM
Gijsbert Frans Dirks
Wouter Olthuis
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
TECHNOLOGIESTICHTING STW
Original Assignee
TECHNOLOGIESTICHTING STW
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 TECHNOLOGIESTICHTING STW filed Critical TECHNOLOGIESTICHTING STW
Priority to AU26438/99A priority Critical patent/AU2643899A/en
Publication of WO1999044046A1 publication Critical patent/WO1999044046A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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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

Definitions

  • the present invention relates to a sensor device for measuring the concentration of a substance, for instance a protein, present in a fluid according to a method wherein this substance is brought as an ionizable analyte into contact with sensor means, comprising a first reagent container for a first reagent which on contact with the analyte causes a first change in electrical charge of the analyte, sensor means for detecting a change in the electrical charge of the analyte and a sample container for holding a sample of the fluid and bringing into contact with the sensor means the substance present in this fluid in the form of an ionizable analyte .
  • analyte is here understood to mean in principle any substance of which the molecules contain an electrically charged group or an ionizable group, such as proteins, or of which the molecules allow linking, for instance by cross-linking, to a polyelectrolyte functioning as so-called charge label, such as heparin or polylysine.
  • Ionizable analyte is understood to mean an analyte of which the electrical charge can be influenced.
  • Such a sensor is for instance important for determining the concentration of particular proteins in human or animal blood.
  • Known from the American patent no. 4.151.049 is a device in which an electrode coated with a layer of a reactive polymer composition specific to a determined protein forms in co-action with a reference electrode an electrically sensitive system for measuring a change in the concentration of that protein in solution. Operation by a highly trained person, for instance a laboratory technician, is a prerequisite to enable use of the known device for determining a protein concentration in a blood sample, and this of course has a cost-increasing effect on such a determination. Application of the known device is further limited to a laboratory environment .
  • Another object is to provide a sensor device which can also be used on a large scale outside a laboratory, for instance at a hospital bed, in a general medical practice, in the home care sector, in situations requiring first aid or in determining contaminants in air, water or ground.
  • Another object is to provide a sensor device which has a higher accuracy than the known device.
  • Providing a second reagent container in a sensor device according to the invention makes this device suitable for performing dynamic measurements, whereby the sensitivity is greatly increased relative to the static measurement which can be performed with a sensor device having only one reagent container.
  • a dynamic measurement in a sensor device according to the invention an analyte system on a sensor surface is stimulated by for instance a change in concentration of an electrolyte and the response to this stimulus is measured. The amplitude of this response is related to the type and concentration of the immobilized analyte .
  • An advantageous embodiment of a sensor device comprises a cartridge which is suitable for insertion into a contact device provided with connection contacts for the sensor means.
  • a sensor device comprising a cartridge makes it possible for filling of the sample container (for instance with a drop of blood from a bedridden patient) and the analysis of that sample (for instance in a hospital laboratory) to be performed in simple manner by the same person not trained in laboratory techniques .
  • first and second reagent containers can be opened (preferably individually) by exerting a determined pressure thereon.
  • the sensor means in a sensor device according to the invention comprise a potentiometric sensor.
  • this potentiometric sensor comprises an ion-sensitive field transistor (ISFET) , the surface of which can be modified by absorption thereon of analyte, and the first and the second reagent are chosen such that on contact with the analyte they cause a different change in electrical charge of the analyte .
  • ISFET ion-sensitive field transistor
  • This latter sensor device is preferably adapted to place the first and the second reagent into alternating contact with the ISFET in order to cause respective different changes in electrical charge of the analyte for measuring by the ISFET.
  • the first and the second reagent for instance contain respectively a first and a second analyte of different concentration.
  • the alternating contact of analyte from the sample container with respectively the first and the second reagent causes in each case a so-called ion step in the ISFET.
  • the former further comprises a reference electrode. So as to ensure that the voltage on this reference electrode is not influenced by an ion step, the first and the second electrolyte in an advantageous embodiment contain different components .
  • a sensor device preferably comprises an overflow container for fluid.
  • a fluid sample or components thereof and excess first and second reagent are collected in this overflow container, so that the carrier respectively the cartridge remain clean and dry during reading of the sensor means and the sample fluid, for instance a blood sample, cannot come into contact with the person carrying out the relevant measurement .
  • the senor means comprise an ISFET
  • they further comprise a reference electrode which in an advantageous embodiment is provided in an overflow container for sample fluid.
  • the invention further relates to a contact device for an above described sensor device, which contact device is provided according to the invention with actuator means for bringing the first respectively the second reagent into contact with analyte placed in contact with the sensor means . 5
  • the contact device is provided with pressing means for exerting a determined pressure on the first and second reagent container.
  • pressing means comprise for instance a pressing roller movable over the surface of the cartridge or a pressing stamp placeable on the surface of the cartridge.
  • the actuator means in the contact device are adapted to individually place the first respectively the second reagent into contact with fluid in the sample container.
  • the contact device is provided with contact means for an ion-sensitive field effect transistor (ISFET) , wherein the actuator means are for instance adapted to bring the first and the second reagent into alternating contact with analyte placed in contact with the sensor means .
  • ISFET ion- sensitive field effect transistor
  • a cartridge according to the invention can be manufactured in simple manner from sheet materials such as cardboard, plastics or laminates, wherein the reagent containers can be formed from plastic foils in a manner which is per se known from the manufacture of reservoirs for developer for instant photographic paper or so-called plastic "blister" materials known from the packaging industry.
  • a thus manufactured cartridge for once-only use is for instance suitable for, but not limited to, performing medical, bio-technological, environmental or veterinary determinations.
  • FIG. 1 shows in perspective view a first embodiment of a cartridge and a schematic representation of a reading device according to the invention
  • Fig. 2 shows in top view a second embodiment of a cartridge according to the invention
  • Fig. 3 shows a longitudinal section of the cartridge of Fig. 2 inserted into a second embodiment of a reading device under a pressing stamp
  • Fig. 4 shows a cross-section of the cartridge of Fig. 2 inserted into a second embodiment of a reading device
  • Fig. 5 is a top view of a third embodiment of a cartridge according to the invention
  • Fig. 6 is a graphic representation of the progression of the current of the drive motor of the pressing stamp of Fig. 3 as a function of time during driving of this stamp.
  • Fig. 1 shows a cartridge 1 and a reading apparatus 3 suitable therefore .
  • a first reservoir 4 filled with a solution of KCl (concentration lOmM) and a second reservoir 8, filled with a solution of KCl (concentration 10 mM) and KN0 3 (concentration 90 mM) , each manufactured from a flexible foil material and closed at the location of respective channels 9, 10 by respective welds 11, 12 which burst open when a pressure is exerted on the reagents in reservoirs 4, 8.
  • Channels 9, 10 are formed by carrier plate 7, intermediate segments 13 arranged thereon and a cover plate
  • the schematically shown reading apparatus 3 comprises pressing rollers 17, 17' and 18, 18' for respectively the first 4 and second reservoir 8 and contact strips 14' -16' which correspond with the respective contact strips 14-16 on cartridge 1.
  • the shown cartridge 1 can for instance be used as immunosensor, suitable for determining an antigen concentration.
  • an antibody protein
  • ligand protein
  • a sample with antigens, the concentration of which must be determined, is introduced into sample container 6.
  • Cartridge 1 is subsequently pushed into reading apparatus 3 in the direction of arrow 19.
  • a contact is first created herein between contact strips 14-16 and the corresponding contact strips 14 ' -16', the weld 11 of the first reservoir 4 is then opened under pressure of pressing rollers 17, 17' and the first reagent flows out of first reservoir 4 via channel 9 to ISFET 5, whereafter weld 12 of second reservoir 8 is opened under pressure of pressing rollers 18, 18' and the second reagent flows out of second reservoir 8 via channel 10 to ISFET 5.
  • the concentration of the sample with antigens can subsequently be determined from a measurement of the response of ISFET 5 to the presented step in ion concentration of the reagents.
  • the response of ISFET 5 is a result of the temporary change in acidity (pH) of the membrane as a consequence of a so-called proton release of titratable groups of the protein molecules immobilized on the ISFET. This rapid and temporary pH change is measured by ISFET 5 with integrated reference electrode.
  • Ligand/analyte couples which display a suitable stimulus response for instance comprise progesterone/anti-progesterone and protamine/heparin (provided with a suitable charge label) .
  • a ligand required for the determination of an analyte does not always have to be immobilized on a membrane, but that some ligands, for instance the protein protamine, can be immobilized directly on the surface of an ISFET.
  • Protamine is for instance suitable as ligand in a sensor device for the determination of the concentration of heparin in a blood sample.
  • Fig. 2 shows a second embodiment of a cartridge 2.
  • a carrier plate 7 manufactured from polycarbonate and having an area of about 60 x 70 mm are a first reservoir 4, filled with a solution of KCl (concentration 10 mM) , and a second reservoir 8, filled with 8 a solution of KCl (concentration 11.3 mM) and KN0 3 (concentration 88.7 mM) , each manufactured from a flexible foil material and closed at the location of respective channels 9, 10 by respective welds 11, 12 which burst open when a pressure is exerted on the reagents in reservoirs 4, 8.
  • Channels 9, 10, which are formed by carrier plate 7, intermediate segments 13 arranged thereon and a cover plate (26, shown in fig.
  • ISFET 5 has a cross-section of about 1000 x 150 ⁇ m and lead to an ISFET 5.
  • ISFET 5 is manufactured in a per se known manner with a gate of Ta 2 0 5 provided with contact strips 20, 21 for respectively the source and the drain of ISFET 5 and is fixed onto carrier plate 7 with epoxy resin in a manner such that the gate is placed with its largest dimension transversely of the direction of flow of sample fluid and reagents .
  • Formed in the cover plate is an opening which provides access to a sample container 6 above ISFET 5.
  • Sample container 6 is closed on its underside by a membrane 32 of nitrocellulose which acts as filter.
  • Overflow channel 22 leads to an overflow reservoir 24 provided with a venting opening 23 and having an Ag/AgCl reference electrode 25 present therein.
  • reference electrode 25 can be placed by way of alternative in one of the channels 9, 10 in which the concentration of electrolyte is constant.
  • Fig. 3 is a longitudinal section of cartridge 2 of Fig. 2 along the line III-III, shown in a situation in which cartridge 2 is inserted into a second embodiment of a reading device under a pressing stamp 27. Shown is how the first reservoir 4 is adhered fixedly between carrier plate 7 and cover plate 26 at the position of weld 11 using a silicone rubber 28, which plates 6, 27 are separated by intermediate segments 13 and between which are formed spaces for an overflow reservoir 24 and (not shown in this section) channels 9, 10 and sample container 6.
  • Fig. 4 is a cross-section of cartridge 2 of Fig. 2 along the line IV-IV, shown in a situation in which cartridge 2 is inserted into a second embodiment of a reading device under contact pins 20', 21' for the corresponding contact strips 20, 21 of respectively the source and drain of ISFET 5. Shown is how ISFET 5 is fixed in a cavity in the polycarbonate carrier plate 7 using an epoxy resin 30.
  • the reading apparatus for the cartridge 2 shown in figures 2-4 contains two stamps (27, Fig. 3) driven by a direct-current motor, and an interface.
  • the motor current is measured in order to enable detection of the opening of reservoirs 4, 8 and to enable regulation of the fluid flows.
  • the need for pressure and conductivity sensors is hereby dispensed with.
  • Both the angular speed of the motor (which is a measure for the fluid flow) and the torque (which is a measure for the pressure exerted by the stamp) can be derived from the motor current and voltage.
  • a first stamp 27 is pressed downwards as according to arrow 29 (Fig. 3) in order to open the first reservoir 4.
  • Bursting open of the first weld 11 is detected as a sudden drop in the motor current caused by a sudden decrease in the load. This sudden current drop is converted by a band-pass filter into a voltage peak which actuates a Schmitt trigger which in turn operates a relay to switch off the motor.
  • both reservoirs 4, 8 are opened and stamps 27 have been stopped, the channels 9, 10 are completely filled with fluid from reservoirs 4, 8.
  • the load which stamps 27 undergo from that moment is practically constant. As a result it is possible to suffice with a control system for displacement of stamps 27 so as to ensure that the fluid flow rate for each subsequent ion step for performing is constant .
  • the interface in the reading apparatus comprises the amplifiers for the drive motors for stamps 27, the ⁇ ⁇ to t M H c ⁇ o c ⁇ o en O in rt CQ rt to CQ rt PJ ⁇ 0 rt ⁇ PJ •d PJ rt •n 0 rt CQ ⁇ ⁇ cr 3 H- ⁇ 3 ⁇ ⁇ ⁇ . ft L ⁇ rt tJ ft ft

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

Abstract

On décrit un dispositif de détection pour mesurer la concentration d'une substance (une protéine, par exemple) présente dans un liquide, suivant un procédé au moyen duquel cette substance est placée, comme analysat ionisable, au contact d'un détecteur. Le dispositif comprend une cartouche munie d'un support sur lequel est placé un premier contenant de réactif pour un premier réactif qui, au contact de l'analysat, induit un premier changement dans la charge électrique de l'analysat, un capteur potentiométrique pour détecter un changement dans la charge électrique de l'analysat, un contenant d'échantillon pour contenir un échantillon du liquide et amener au contact du capteur la substance présente dans le liquide sous la forme d'un analysat ionisable, et un deuxième contenant de réactif pour un deuxième réactif qui, au contact de l'analysat, induit un deuxième changement dans la charge électrique de l'analysat. Le premier et le deuxième contenants de réactif peuvent s'ouvrir individuellement lorsqu'on y exerce une pression déterminée, et la cartouche peut s'insérer efficacement dans un dispositif de contact muni de contacts de connexion pour le capteur, et un dispositif de contact pour ledit dispositif de détection.
PCT/NL1999/000085 1998-02-25 1999-02-17 Cartouche equipee d'un detecteur de proteine et dispositif de lecture associe Ceased WO1999044046A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU26438/99A AU2643899A (en) 1998-02-25 1999-02-17 Cartridge with protein-sensor and reading device therefore

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
NL1008411 1998-02-25
NL1008411A NL1008411C2 (nl) 1998-02-25 1998-02-25 Cassette met proteïne-sensor en uitleesinrichting daarvoor.

Publications (1)

Publication Number Publication Date
WO1999044046A1 true WO1999044046A1 (fr) 1999-09-02

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PCT/NL1999/000085 Ceased WO1999044046A1 (fr) 1998-02-25 1999-02-17 Cartouche equipee d'un detecteur de proteine et dispositif de lecture associe

Country Status (3)

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AU (1) AU2643899A (fr)
NL (1) NL1008411C2 (fr)
WO (1) WO1999044046A1 (fr)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1114995A3 (fr) * 1999-12-30 2003-10-15 Roche Diagnostics Corporation Cellule et méthode pour l'analyse électrochimique d'un échantillon
WO2005066275A3 (fr) * 2003-12-23 2005-09-01 Gen Electric Dispositifs capteurs contenant des substrats a base de copolymeres pour l'analyse d'especes chimiques et biologiques dans l'eau et dans l'air
EP1392860A4 (fr) * 2001-04-23 2006-11-29 Samsung Electronics Co Ltd Puce de detection moleculaire comprenant un transistor mosfet, dispositif de detection moleculaire utilisant ladite puce et procede de detection moleculaire faisant appel au dit dispositif
RU2377571C2 (ru) * 2005-01-07 2009-12-27 Секисуй Кемикал Ко., Лтд. Детекторное устройство с использованием картриджа

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0127958A2 (fr) * 1983-05-05 1984-12-12 MediSense, Inc. Electrodes pour des détecteurs
EP0306158A2 (fr) * 1987-08-29 1989-03-08 THORN EMI plc Cartouche avec capteur
WO1990002938A1 (fr) * 1988-09-15 1990-03-22 I-Stat Corporation Dispositif de detection jetable pour l'analyse de fluides en temps reel
WO1991009373A1 (fr) * 1989-12-15 1991-06-27 Boehringer Mannheim Corporation Machine et instrument de biodetection
GB2293883A (en) * 1994-06-30 1996-04-10 Nok Corp Electrochemical protein biosensor

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0127958A2 (fr) * 1983-05-05 1984-12-12 MediSense, Inc. Electrodes pour des détecteurs
EP0306158A2 (fr) * 1987-08-29 1989-03-08 THORN EMI plc Cartouche avec capteur
WO1990002938A1 (fr) * 1988-09-15 1990-03-22 I-Stat Corporation Dispositif de detection jetable pour l'analyse de fluides en temps reel
WO1991009373A1 (fr) * 1989-12-15 1991-06-27 Boehringer Mannheim Corporation Machine et instrument de biodetection
GB2293883A (en) * 1994-06-30 1996-04-10 Nok Corp Electrochemical protein biosensor

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1114995A3 (fr) * 1999-12-30 2003-10-15 Roche Diagnostics Corporation Cellule et méthode pour l'analyse électrochimique d'un échantillon
EP1392860A4 (fr) * 2001-04-23 2006-11-29 Samsung Electronics Co Ltd Puce de detection moleculaire comprenant un transistor mosfet, dispositif de detection moleculaire utilisant ladite puce et procede de detection moleculaire faisant appel au dit dispositif
US7235389B2 (en) 2001-04-23 2007-06-26 Samsung Electronics Co., Ltd. Molecular detection device and chip including MOSFET
US7781167B2 (en) 2001-04-23 2010-08-24 Samsung Electronics Co., Ltd. Molecular detection methods using molecular detection chips including a metal oxide semiconductor field effect transistor
US7863140B2 (en) 2001-04-23 2011-01-04 Samsung Electronics Co., Ltd. Methods of making a molecular detection chip having a metal oxide silicon field effect transistor on sidewalls of a micro-fluid channel
WO2005066275A3 (fr) * 2003-12-23 2005-09-01 Gen Electric Dispositifs capteurs contenant des substrats a base de copolymeres pour l'analyse d'especes chimiques et biologiques dans l'eau et dans l'air
RU2377571C2 (ru) * 2005-01-07 2009-12-27 Секисуй Кемикал Ко., Лтд. Детекторное устройство с использованием картриджа

Also Published As

Publication number Publication date
NL1008411C2 (nl) 1999-08-26
AU2643899A (en) 1999-09-15

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