WO2002088738A2 - Device and method for the assay of a specific particle in a liquid sample - Google Patents

Device and method for the assay of a specific particle in a liquid sample Download PDF

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Publication number
WO2002088738A2
WO2002088738A2 PCT/NL2002/000172 NL0200172W WO02088738A2 WO 2002088738 A2 WO2002088738 A2 WO 2002088738A2 NL 0200172 W NL0200172 W NL 0200172W WO 02088738 A2 WO02088738 A2 WO 02088738A2
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WO
WIPO (PCT)
Prior art keywords
specific type
particles
binding
liquid
liquid sample
Prior art date
Application number
PCT/NL2002/000172
Other languages
French (fr)
Other versions
WO2002088738A3 (en
Inventor
Richard Bernardus Maria Schasfoort
Original Assignee
Ibis Technologies B.V.
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 Ibis Technologies B.V. filed Critical Ibis Technologies B.V.
Priority to AU2002239166A priority Critical patent/AU2002239166A1/en
Publication of WO2002088738A2 publication Critical patent/WO2002088738A2/en
Publication of WO2002088738A3 publication Critical patent/WO2002088738A3/en

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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
    • G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
    • G01N33/585—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with a particulate label, e.g. coloured latex
    • G01N33/587—Nanoparticles
    • 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 device for the assay of at least one specific type of particle in a liquid sample.
  • the invention also relates to a method for the assay of at least one specific type of particle in a liquid sample by means of such a device.
  • Bio-diagnostic techniques based on affinity reactions are known in the art, e.g. antibody/antigen and receptor/ligand reactions. These techniques are used for biochemical analyses of samples of body liquids such as blood, serum and urine.
  • the 'sandwich method' is often used, whereby a ligand is adsorbed onto a fixed carrier.
  • the particle to be assayed specifically binds to the ligand.
  • a conjugate is added that in turn binds specifically to the particle to be assayed.
  • This conjugate in turn can be linked to a fluorescent particle.
  • the presence of the particle can be dete ⁇ nined, e.g by means of a fluorescence microscope.
  • the purpose of the present invention is to provide a device and a method with which a more complicated bio-diagnostic assay with all necessary steps can be performed automatically according to an optimised process in relatively little time, with only relatively simple operations.
  • the invention provides a device for the assay of at least one specific type of particle in a liquid sample, the device comprising:
  • a hydraulic network comprising at least one capillary duct and at least one liquid reservoir; - at least two electrodes for the application of an electrical potential difference for the purpose of generating an electro-osmotic flow in at least a part of the hydraulic network;
  • the necessary liquids such as the liquid sample, rinsing liquid, conjugate and waste liquid can be stored separately in the reservoirs.
  • the liquid movements to and from the binding surface, from and to the reservoirs and through the ducts can be controlled by the application of electrical potential differences by means of the electrodes.
  • the whole of hydraulic network with inlets and outlets, electrodes and binding surface can be manufactured integrated and miniaturised by means of methods known from the fields of 'precision technology' and 'microsystem technology'.
  • the capillary duct is made in a substrate and has a width and depth of between 1 and 1,000 ⁇ m and between 1 and 1,000 ⁇ m respectively, preferably between 10 and 100 ⁇ m and 10 and 100 ⁇ m respectively.
  • the substrate can be glass or a plastic such as PMMA or polystyrene.
  • the making of the ducts can be carried out by means of an etching technique, an abrasive technique or a technique like hot-embossing, possibly in combination with a photolithographical technique.
  • a relatively complex hydraulic network of capillary ducts can be manufactured in an integrated fashion.
  • the walls of the ducts can optionally be provided with a suitable coating.
  • the dimensions of a duct should not be too large so as to be able to generate sufficient electro-osmotic flow, or too small for sufficient throughput.
  • the invention also provides a method for the assay of at least one specific type of particle in a liquid sample using a device according to the invention, the method comprising the following steps:
  • a biochemical analysis based on an affinity reaction can be performed automatically and in an optimised way, whereby the liquid movements from and to the binding surface, from and to the reservoirs and through the ducts can be programmed, and directed by applying electrical potential differences by means of the electrodes on the appropriate places and moments.
  • the liquid sample is a body liquid, and the specific type of particle a bio-organic molecule present in the body liquid.
  • the specific type of particle a bio-organic molecule present in the body liquid.
  • Use is made of e.g. an antibody/antigen or receptor/ligand reaction.
  • the method can also comprise the step: X. application of a marker on at least a part of the specific particles bound to the binding sites on the binding surface.
  • the specific type of particles bound to the binding surface can be better and more easily detected and assayed.
  • the marker is fluorescent.
  • the particles of the specific type bound to the binding surface, provided with such a marker can thus be made easily visible and be assayed by means of e.g. fluorescence imaging microscopy.
  • the fluorescent marker can be a latex sphere, preferably a polystyrene latex one, preferably with a diameter of between 1 and 100,000 nm, preferably between 10 and 10,000 nm.
  • a fluorescent marker proves to be suitable.
  • FIG. 1 shows a schematic top view of a part of a preferred embodiment of a device according to the invention.
  • Fig.l shows a 'chip' 1 provided with a network of capillary ducts 2.
  • the reservoirs 3-5 contain a blood sample 6, a rinsing liquid 7 and a fluorescent conjugate 8, respectively.
  • Reservoir 9 serves for the storage of waste liquid 10.
  • a part 11 of the chip 1 serves as a 'detection surface'.
  • a part 12 of this has been provided with a coating that constitutes a fixed carrier.
  • On a part 13 of this a suitable ligand has been adsorbed in order to provide a number of binding sites.
  • a part 14 of the detection surface 11 has not been provided with a coating and serves as a reference.
  • Part of the blood sample 6 is directed through the capillary duct network 2 by means of electro-osmosis to the detection surface 11.
  • part 13 a number of the particles to be assayed, if present in the blood sample 6, will bind to the ligand on the binding sites.
  • part of the rinsing liquid 7 is directed from reservoir 4 along the detection surface 11 and disposed of in reservoir 9.
  • the fluorescent conjugate 8 from reservoir 5 is directed past the detection surface 11 and disposed of in reservoir 9.
  • the entire detection surface 11 can be inspected with a fluorescence imaging microscope. If the particle to be assayed is present the part 13 of the detection surface will light up. Thus the presence of the particle concerned in the blood sample 6 can be demonstrated.
  • use can be made of the other parts 11, 12 and 14 as a reference.
  • the detection surface consist of a number of parts that are used as a reference

Abstract

A device and method for the assay of at least one specific type of particle in a liquid sample. The device comprising: - a hydraulic network comprising at least one capillary duct and at least one fluid reservoir; - at least two electrodes for the application of an electric potential difference for the purpose of generating an electro-osmotic flow in at least a part of the hydraulic network; - at least one binding surface connected to the hydraulic network along which surface a liquid can flow, the surface being suitable to be provided with binding sites where a selective bonding can take place with particles of the specific type, and - a detection system for the assaying of the particles of the specific type. The method comprising the following steps: A. the application of a number of binding sites onto the binding surface; B. conducting of at least a part of the liquid sample to the binding surface by generating an electro-osmotic flow in at least a part of the hydraulic network by the application of electric potential differences by means of electrodes, whereby a bonding takes place between particles of the specific type and at least a part of the binding sites applied on the binding surface, and C. assaying the particles of the specific type by means of a detection system.

Description

Device and method for the assay of at least one specific type of particle in a liquid sample
Field of the Invention
The present invention relates to a device for the assay of at least one specific type of particle in a liquid sample. The invention also relates to a method for the assay of at least one specific type of particle in a liquid sample by means of such a device.
Description of the Prior Art
Bio-diagnostic techniques based on affinity reactions are known in the art, e.g. antibody/antigen and receptor/ligand reactions. These techniques are used for biochemical analyses of samples of body liquids such as blood, serum and urine.
The 'sandwich method' is often used, whereby a ligand is adsorbed onto a fixed carrier. The particle to be assayed specifically binds to the ligand. After rinsing, a conjugate is added that in turn binds specifically to the particle to be assayed. This conjugate in turn can be linked to a fluorescent particle. After a further rinse, the presence of the particle can be deteπnined, e.g by means of a fluorescence microscope.
The necessary number of steps for incubation, concentration and rinsing with the abovementioned diagnostical methods is often large. Consequently, the assays often are complex and time consuming, and for a correct execution skilled staff is needed. For certain applications when a quick result is desired, e.g. in a first aid centre, an ambulance or during an operation, or when an unskilled person should be able to perform the assay, the 'dipstick' method has been developed. The required liquid transport takes place through capillary movement whereby a number of subsequent reactions are passed automatically. A known example is the pregnancy test, whereby the dipstick only needs to be dipped in the urine, after which the result can be read from local discolorations. For more complicated assays with relatively many steps and liquid movements this dipstick method is not adequate. Summary of the Invention
The purpose of the present invention is to provide a device and a method with which a more complicated bio-diagnostic assay with all necessary steps can be performed automatically according to an optimised process in relatively little time, with only relatively simple operations.
To this end, the invention provides a device for the assay of at least one specific type of particle in a liquid sample, the device comprising:
- a hydraulic network comprising at least one capillary duct and at least one liquid reservoir; - at least two electrodes for the application of an electrical potential difference for the purpose of generating an electro-osmotic flow in at least a part of the hydraulic network;
- at least one binding surface connected to the hydraulic network along which surface a liquid can flow, the surface being suitable to be provided with binding sites where a selective bonding can take place with particles of the specific type, and
- a detection system for the assaying of the particles of the specific type.
The necessary liquids such as the liquid sample, rinsing liquid, conjugate and waste liquid can be stored separately in the reservoirs. The liquid movements to and from the binding surface, from and to the reservoirs and through the ducts can be controlled by the application of electrical potential differences by means of the electrodes. The whole of hydraulic network with inlets and outlets, electrodes and binding surface can be manufactured integrated and miniaturised by means of methods known from the fields of 'precision technology' and 'microsystem technology'. Preferably the capillary duct is made in a substrate and has a width and depth of between 1 and 1,000 μm and between 1 and 1,000 μm respectively, preferably between 10 and 100 μm and 10 and 100 μm respectively.
The substrate can be glass or a plastic such as PMMA or polystyrene. The making of the ducts can be carried out by means of an etching technique, an abrasive technique or a technique like hot-embossing, possibly in combination with a photolithographical technique. Thus a relatively complex hydraulic network of capillary ducts can be manufactured in an integrated fashion. The walls of the ducts can optionally be provided with a suitable coating. The dimensions of a duct should not be too large so as to be able to generate sufficient electro-osmotic flow, or too small for sufficient throughput. The invention also provides a method for the assay of at least one specific type of particle in a liquid sample using a device according to the invention, the method comprising the following steps:
A. the application of a number of binding sites onto the binding surface; B. conducting of at least a part of the liquid sample to the binding surface by generating an electro-osmotic flow in at least a part of the hydraulic network by the application of electrical potential differences by means of the electrodes, whereby a bonding takes place between particles of the specific type and at least a part of the binding sites applied on the binding surface, and C. assaying the particles of the specific type by means of a detection system.
With this method a biochemical analysis based on an affinity reaction can be performed automatically and in an optimised way, whereby the liquid movements from and to the binding surface, from and to the reservoirs and through the ducts can be programmed, and directed by applying electrical potential differences by means of the electrodes on the appropriate places and moments.
Preferably the liquid sample is a body liquid, and the specific type of particle a bio-organic molecule present in the body liquid. Use is made of e.g. an antibody/antigen or receptor/ligand reaction.
At that, the method can also comprise the step: X. application of a marker on at least a part of the specific particles bound to the binding sites on the binding surface.
By applying a marker the specific type of particles bound to the binding surface can be better and more easily detected and assayed.
Preferably the marker is fluorescent. The particles of the specific type bound to the binding surface, provided with such a marker can thus be made easily visible and be assayed by means of e.g. fluorescence imaging microscopy.
Here the fluorescent marker can be a latex sphere, preferably a polystyrene latex one, preferably with a diameter of between 1 and 100,000 nm, preferably between 10 and 10,000 nm. In practice such a fluorescent marker proves to be suitable. Preferably use is made of surface plasmon resonance for assaying the particles of the specific type by means of the detection system. This is a very sensitive method that is very well suited to detection and assaying of particles bound to a surface. Description of a Preferred Embodiment
Hereafter the present invention will be illustrated by means of an non-limitative example. To this end Fig. 1 shows a schematic top view of a part of a preferred embodiment of a device according to the invention.
Fig.l shows a 'chip' 1 provided with a network of capillary ducts 2. The reservoirs 3-5 contain a blood sample 6, a rinsing liquid 7 and a fluorescent conjugate 8, respectively. Reservoir 9 serves for the storage of waste liquid 10. A part 11 of the chip 1 serves as a 'detection surface'. A part 12 of this has been provided with a coating that constitutes a fixed carrier. On a part 13 of this a suitable ligand has been adsorbed in order to provide a number of binding sites. A part 14 of the detection surface 11 has not been provided with a coating and serves as a reference.
Part of the blood sample 6 is directed through the capillary duct network 2 by means of electro-osmosis to the detection surface 11. In part 13 a number of the particles to be assayed, if present in the blood sample 6, will bind to the ligand on the binding sites. Next, part of the rinsing liquid 7 is directed from reservoir 4 along the detection surface 11 and disposed of in reservoir 9. After this the fluorescent conjugate 8 from reservoir 5 is directed past the detection surface 11 and disposed of in reservoir 9. After a further rinse the entire detection surface 11 can be inspected with a fluorescence imaging microscope. If the particle to be assayed is present the part 13 of the detection surface will light up. Thus the presence of the particle concerned in the blood sample 6 can be demonstrated. For a quantitative analyse use can be made of the other parts 11, 12 and 14 as a reference.
Special characteristics and advantages of this device and this method are among others: - the ensemble of ducts and detection surface has been integrally made in and on a substrate;
- the liquid movements from and to the detection surface, from and to the reservoirs and in both directions in the ducts, can be controlled ad lib by the application of electrical potential differences by means of the electrodes on appropriate places and moments; - a complete optimised bio-diagnostic assay with all necessary steps can be preprogrammed and be executed automatically;
- the detection surface consist of a number of parts that are used as a reference, and
- the complete process can be observed and studied with a fluorescence microscope.

Claims

Claims
1. Device for the assay of at least one specific type of particle in a liquid sample, the device comprising: - a hydraulic network comprising at least one capillary duct and at least one liquid reservoir;
- at least two electrodes for the application of an electrical potential difference for the purpose of generating an electro-osmotic flow in at least a part of the hydraulic network; - at least one binding surface connected to the hydraulic network along which surface a liquid can flow, the surface being suitable to be provided with binding sites where a selective bonding can take place with particles of the specific type, and
- a detection system for the assaying of the particles of the specific type.
2. A device according to claim 1, characterised by the capillary duct being made in a substrate, having a width and depth of between 1 and 1,000 μm, and 1 and 1,000 μm respectively, preferably between 10 and 100 μm and 1 and 100 μm respectively.
3. A method for the assay of at least one specific type of particle in a liquid sample by means of a device according to one of the preceding claims, comprising the following steps:
A. the application of a number of binding sites onto the binding surface;
B. conducting of at least a part of the liquid sample to the binding surface by generating an electro-osmotic flow in at least a part of the hydraulic network by the application of electrical potential diffferences by means of the electrodes, whereby a bonding takes place between particles of the specific type and at least a part of the binding sites applied on the binding surface, and
C. assaying the particles of the specific type by means of a detection system.
4. A method according to claim 3, characterised by the fact that the liquid sample is a body liquid, and the specific type of particle is a bio-organic molecule present in the body liquid.
5. A method according to claim 3 or 4, characterised by the fact that the method also comprises the step:
X. application of a marker on at least a part of the specific particles bound to the binding sites on the binding surface.
6. A method according to claim 5, characterised by the fact that the marker is fluorescent.
7. A method according to claim 6, characterised by the fact that the fluorescent marker is a latex sphere, preferably a polystyrene latex sphere, preferably with a diameter between 1 and 100,000 nm, preferably between 10 and 10,000 nm.
8. A method according to one of the conclusions 3-7, characterised by the fact that for the assay of particles of the specific type by means of the detection system use is being made of surface plasmon resonance.
PCT/NL2002/000172 2001-03-16 2002-03-13 Device and method for the assay of a specific particle in a liquid sample WO2002088738A2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU2002239166A AU2002239166A1 (en) 2001-03-16 2002-03-13 Device and method for the assay of a specific particle in a liquid sample

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
NL1017625 2001-03-16
NL1017625A NL1017625C2 (en) 2001-03-16 2001-03-16 Device and method for determining at least one specific type of particle in a liquid sample.

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WO2002088738A2 true WO2002088738A2 (en) 2002-11-07
WO2002088738A3 WO2002088738A3 (en) 2003-05-08

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NL (1) NL1017625C2 (en)
WO (1) WO2002088738A2 (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
NL1027633C2 (en) * 2004-12-01 2006-06-02 Stichting Tech Wetenschapp Determining several types of target particles in a sample comprises loading zones of a carrier with different types of first particles and contacting subzones with mixtures of second particles bound to third particles

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5194300A (en) * 1987-07-15 1993-03-16 Cheung Sau W Methods of making fluorescent microspheres
WO1996002837A1 (en) * 1994-07-20 1996-02-01 Sios, Inc. Apparatus and method for the detection and assay of organic molecules
EP0851230A1 (en) * 1996-12-30 1998-07-01 Diagnostic Products Corporation Method and Apparatus for Immunoassay using Fluorescent Induced Surface Plasma Emission

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5194300A (en) * 1987-07-15 1993-03-16 Cheung Sau W Methods of making fluorescent microspheres
WO1996002837A1 (en) * 1994-07-20 1996-02-01 Sios, Inc. Apparatus and method for the detection and assay of organic molecules
EP0851230A1 (en) * 1996-12-30 1998-07-01 Diagnostic Products Corporation Method and Apparatus for Immunoassay using Fluorescent Induced Surface Plasma Emission

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
NL1027633C2 (en) * 2004-12-01 2006-06-02 Stichting Tech Wetenschapp Determining several types of target particles in a sample comprises loading zones of a carrier with different types of first particles and contacting subzones with mixtures of second particles bound to third particles

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AU2002239166A1 (en) 2002-11-11
WO2002088738A3 (en) 2003-05-08
NL1017625C2 (en) 2002-09-24

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