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WO2003012389A2 - Procede pour analyser des macromolecules, dispositif d'analyse et procede pour produire un dispositif d'analyse - Google Patents

Procede pour analyser des macromolecules, dispositif d'analyse et procede pour produire un dispositif d'analyse Download PDF

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Publication number
WO2003012389A2
WO2003012389A2 PCT/EP2002/007644 EP0207644W WO03012389A2 WO 2003012389 A2 WO2003012389 A2 WO 2003012389A2 EP 0207644 W EP0207644 W EP 0207644W WO 03012389 A2 WO03012389 A2 WO 03012389A2
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WIPO (PCT)
Prior art keywords
macromolecules
liquid
drop
spots
liquid drop
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PCT/EP2002/007644
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German (de)
English (en)
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WO2003012389A3 (fr
Inventor
Jürgen SCRIBA
Christoph Gauer
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Advalytix Ag
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Application filed by Advalytix Ag filed Critical Advalytix Ag
Priority to AU2002328867A priority Critical patent/AU2002328867A1/en
Priority to EP02764659A priority patent/EP1412533A2/fr
Publication of WO2003012389A2 publication Critical patent/WO2003012389A2/fr
Publication of WO2003012389A3 publication Critical patent/WO2003012389A3/fr

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Classifications

    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366Apparatus specially adapted for solid-phase testing
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027Containers 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
    • B01L3/502769Containers 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 characterised by multiphase flow arrangements
    • B01L3/502784Containers 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 characterised by multiphase flow arrangements specially adapted for droplet or plug flow, e.g. digital microfluidics
    • B01L3/502792Containers 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 characterised by multiphase flow arrangements specially adapted for droplet or plug flow, e.g. digital microfluidics for moving individual droplets on a plate, e.g. by locally altering surface tension
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01JCHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00277Apparatus
    • B01J2219/00497Features relating to the solid phase supports
    • B01J2219/00527Sheets
    • B01J2219/00533Sheets essentially rectangular
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01JCHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00583Features relative to the processes being carried out
    • B01J2219/00603Making arrays on substantially continuous surfaces
    • B01J2219/00659Two-dimensional arrays
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01JCHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/0068Means for controlling the apparatus of the process
    • B01J2219/00702Processes involving means for analysing and characterising the products
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00Additional constructional details
    • B01L2300/06Auxiliary integrated devices, integrated components
    • B01L2300/0627Sensor or part of a sensor is integrated
    • B01L2300/0636Integrated biosensor, microarrays
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00Additional constructional details
    • B01L2300/08Geometry, shape and general structure
    • B01L2300/089Virtual walls for guiding liquids
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00Moving or stopping fluids
    • B01L2400/04Moving fluids with specific forces or mechanical means
    • B01L2400/0403Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/0433Moving fluids with specific forces or mechanical means specific forces vibrational forces
    • B01L2400/0436Moving fluids with specific forces or mechanical means specific forces vibrational forces acoustic forces, e.g. surface acoustic waves [SAW]
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00Moving or stopping fluids
    • B01L2400/04Moving fluids with specific forces or mechanical means
    • B01L2400/0403Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/0433Moving fluids with specific forces or mechanical means specific forces vibrational forces
    • B01L2400/0439Moving fluids with specific forces or mechanical means specific forces vibrational forces ultrasonic vibrations, vibrating piezo elements
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00Moving or stopping fluids
    • B01L2400/04Moving fluids with specific forces or mechanical means
    • B01L2400/0493Specific techniques used
    • B01L2400/0496Travelling waves, e.g. in combination with electrical or acoustic forces
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6813Hybridisation assays
    • C12Q1/6834Enzymatic or biochemical coupling of nucleic acids to a solid phase
    • C12Q1/6837Enzymatic or biochemical coupling of nucleic acids to a solid phase using probe arrays or probe chips

Definitions

  • the invention relates to methods for analyzing macromolecules with the aid of a microarray and a method for producing an analysis device for macromolecules.
  • sample molecules A liquid with macromolecules (“sample molecules”) is rinsed over the microarray, which forms a specific bond with at least one type of probe molecule on the microarray (hybridization). If the liquid is then removed from the surface again, the sample molecules to be examined remain only at the points of the specific binding. With the help of a spatially resolved measurement, e.g. B.
  • a fluorescence measurement it can be determined at which locations to be examined macromolecules are present. From the known position of the individual probe molecules in the matrix form of the microarray it can thus be determined with which type of macromolecules the macromolecules to be examined have entered into a specific bond.
  • microarrays are e.g. B. used to study macromolecules such as proteins, antigens or antibodies.
  • microarrays are also used to examine DNA, e.g. B. used in DNA screening.
  • the duration of a corresponding analysis experiment is largely determined by the diffusion of the sample molecules to the probe molecules and can therefore take some time. Is z. For example, if the concentration of the macromolecule to be examined in the liquid is only low, it can take a long time before it has found its specific binding partner on the array.
  • the microarrays used are miniaturized arrays, which in the case of e.g. B. of DNA microarrays include individual areas (spots) with probe molecules each with a diameter of about 100 microns. There are many different DNA probes (spots) z. B. applied in a checkerboard pattern on the surface and simultaneously used for hybridization reactions.
  • the densities that can be achieved on such a microarray today are, for. B. for DNA (deoxyribonucleic acid) up to about 250,000 different types of molecules per cm 2 (M. Schena (ed.), "DNA Microarrays", Oxford University Press, 1999).
  • the microarray is washed after hybridization according to a special protocol and then the fluorescence is measured in a spatially resolved manner.
  • One possibility is to measure the fluorescence signal with optics of such a shallow depth of field that only the area of the interface between the liquid film and the chip surface contributes to the signal.
  • the second macromolecules are mainly in the already bound state there. The fluorescence signal of two macromolecules, which are in the unbound state in the liquid film above them, is not detected with such optics.
  • the probe molecules are mostly applied using a printing technique (spotting).
  • a printing technique is used e.g. B. from VG Cheung et al., Nature Genetic Supplement, Volume 21, January 1999, pages 15ff. described.
  • a solution with the desired probe molecule is brought onto the chip surface either in a contactless or contacting manner.
  • the probe molecules can also be e.g. B. can be synthesized photolithographically base by base on the chip. Both at Spotting as well as in photolithographic production is difficult to make a quantitative statement about the number and density of the probe molecules on a spot. That means that e.g. B. when detecting the hybridized sample molecules z. B. only limited quantitative conclusions about the number of sample molecules are possible by fluorescence. The comparability of data obtained on different microarrays with nominally identical spots is considerably restricted.
  • the surface of a solid state chip is first functionalized homogeneously, so that, for. B. by the action of light or by the action of an electric field bind the previously also functionalized probe molecules on the surface.
  • the chip is homogeneously flooded with a solution of the probe molecules (e.g. in the order of magnitude of 100 ⁇ l).
  • Local illumination binds photoactive probe molecules locally to the desired locations.
  • electroactive probe molecules which bind to the surface under the action of an electrical field
  • an electrical field is generated locally which triggers the binding.
  • the object of the present invention is to provide methods and a device with which an accurate and reproducible analysis of macromolecules is possible, which can also be used with limited sample material.
  • the production of corresponding analysis devices should be simple and reproducible.
  • the object is achieved by a method having the features of claim 1 or a method having the features of claim 6.
  • the object is achieved by a method having the features of claim 15.
  • the analysis device the object is achieved with an analysis device with the features of claim 25.
  • Subclaims are each directed to preferred configurations.
  • a microarray is used on a solid surface, preferably a chip. At least one drop of liquid with the macromolecules to be examined is placed on the surface of the solid, the amount of liquid in a drop being so small that it essentially touches only one spot or a group of a few spots of the microarray at a time. Conveniently, no more than 10% of all spots of a microarray should be touched by a liquid drop. In this way, the reaction conditions (e.g. temperature or salt concentration) can be optimized depending on the spots touched.
  • the reaction conditions e.g. temperature or salt concentration
  • the process is preferably carried out in such a way that only one spot is touched by a drop of liquid at a time, so that the reaction conditions can be set individually for each individual spot.
  • the z. B. is determined by the typical reaction time of the macromolecules contained in the liquid with the macromolecules, which are located as probe molecules on a spot, is moved to a next spot with the aid of the impulse transfer of a surface acoustic wave of the liquid drops.
  • a predetermined time is again waited until the typical reaction time has passed.
  • the desired spots or all spots of the microarray used with the liquid can be sequenced be brought into contact.
  • this process can also be parallelized by moving several drops simultaneously over different spots.
  • the spots that have come into contact with the liquid drop can then be examined for whether or to what extent a reaction with the macromolecules in the liquid drop has taken place.
  • e.g. B. serve a spatially resolved fluorescence measurement if the macromolecules to be examined are fluorescence-labeled.
  • Other detection methods, e.g. B. using radioactive sample molecules can also be used.
  • the examination of the spots that have come into contact with the liquid drop to determine whether and / or to what extent a specific reaction with the material of the liquid drop has taken place can be carried out at the end of the entire process, i. H. when the drop of liquid was in contact with all the desired spots.
  • the process is carried out appropriately, it is also possible, for example, to measure the fluorescence of the bound second macromolecules in situ.
  • optics with shallow depth of field or appropriately arranged waveguides and evanescent waves can be used, which only detect the interface between the liquid and the chip surface.
  • the liquid with the sample molecules is not applied to the entire surface of the microarray, but rather only to individual spots which are separated from one another. While in classic microarray experiments liquid volumes of about 10-100 ⁇ l are necessary to cover a total sample area of 1 cm 2 , the amount of liquid required can be reduced to a few nl with the method according to the invention if, for. B.
  • Spots with an extension of 100 microns can be used.
  • the small sample drop is gradually moved from spot to spot.
  • the diffusion length is not in the order of 1 cm, but only in the order of 100 ⁇ m.
  • a quick and efficient hybridization reaction then takes place at the spots themselves.
  • the concentration of the starting materials can be much higher even with the same amount of starting material, because instead of about 10-100 ⁇ l, for. B. only about 10 nl are needed.
  • the reaction equilibrium can thus be pushed to the side of the hybridized products, so that a much more precise measurement is possible even with little material to be examined.
  • the time that is waited before the liquid drop is removed again from a spot can be shortened by moving the liquid drop within itself by means of the impulse transmission of a surface sound wave, that is to say “stirring” it.
  • a surface sound wave of lower intensity is used for this. than that for transporting the liquid drop so that the liquid drop does not move away from the spot.
  • the method according to the invention allows a freely programmable path of the sample liquid over the surface.
  • an experimental strategy can be followed in order to first approach those spots for which a positive result is expected.
  • the reaction questions can be conditions can be selected differently. So z. B. the temperature can be set differently using resistance heaters.
  • the chip are heated homogeneously, since the spatial resolution is guaranteed by the limited expansion of the drop.
  • the nature of the liquid to be examined can be changed from one spot to the next. While in a spot z. B. the salt concentration in the liquid has a first value, it can be changed to a second value at a second spot by adding appropriate buffer solution.
  • the buffer solution can also be moved in the direction of the corresponding spot with the aid of the impulse transmission of a surface sound wave.
  • the solid surface z. B. is freed from the liquid in a separate washing step before examining whether and to what extent a reaction of the second macromolecules in the liquid with the first macromolecules of the spots has taken place.
  • the liquid drop is moved away from the corresponding spot with the aid of the surface sound wave, so that an investigation can be carried out immediately afterwards to determine whether bound second macromolecules have remained on the spot.
  • the evaluation can take place parallel to the hybridization. For example, a drop is moved from a fluorescence-labeled sample molecule type in the sample liquid to a first spot. After the typical hybridization time, the drop is moved to the next spot and the fluorescence measured in parallel at the first spot. At the second spot z. For example, to change the reaction conditions, change the temperature or add a small volume of buffer solution to ensure the stringency of the reaction.
  • the amount of second macromolecules bound to the respective spot is determined by the damping and / or change in speed of a surface sound wave determines the presence of the second macromolecules.
  • a surface sound wave is sent in the direction of the spot with the second macromolecules bound to the first macromolecule.
  • the mass on the surface which is formed by the hybridized second macromolecules, dampens the surface sound wave in its intensity or changes its speed.
  • a detector for the surface sound wave its intensity can be measured before and after the hybridization of the second macromolecules, and the number of bound second macromolecules can be deduced from the damping or change in speed determined thereby. If necessary, a calibration measurement can be carried out with known parameters.
  • At least one drop of liquid with second macromolecules is brought onto the surface of the solid body with the microarray, the amount of liquid in a drop being so small that essentially only one spot or a group of a few spots of the microarray at a time with the Drops of liquid come into contact.
  • the drop is moved back and forth with the help of the impulse transmission of two essentially opposite surface sound waves, which are sent alternately in the direction of the liquid drop.
  • the intensity of the surface sound waves is chosen so that the drop is not completely removed.
  • the liquid drop is moved with the aid of the impulse transmission of surface sound waves.
  • a surface acoustic wave is generated on the surface of the chip on which the microarray is located.
  • the momentum transfer is then achieved either by the mechanical deformation of the solid surface or by the force effect of the accompanying electric fields on charged or polarizable matter in the liquid.
  • the strength of the force acting on the liquid can be set in a wide range via the amplitude of the surface sound wave.
  • the surface sound wave can be generated in pulses with pulses of different lengths or continuously.
  • Surface acoustic waves can be on piezoelectric substrates or substrates with piezoelectric areas, e.g. B. piezoelectric coatings.
  • the movement with the help of the impulse transfer of surface sound waves is particularly precise.
  • the surface acoustic wave can be generated with the aid of at least one surface wave generating device.
  • An interdigital transducer known per se is advantageously used for this purpose.
  • such an interdigital transducer has two electrodes that interlock like fingers.
  • a high-frequency alternating field e.g. B. in the order of 10 to a few 100 MHz
  • a surface acoustic wave is excited in a piezoelectric substrate or in a piezoelectric region of the substrate, the Wavelength is the quotient of the surface sound velocity and the frequency.
  • the direction of propagation is perpendicular to the interdigitated finger electrode structures.
  • Interdigital transducers can also, e.g. B. can be controlled wirelessly by irradiation of an alternating electromagnetic field in an antenna device connected to the interdigital transducer.
  • interdigital transducers can be used in the desired arrangement, which are selectively controlled.
  • a laterally limited sound path of the surface sound wave is generated with the aid of so-called tapered interdigital transducers.
  • the finger spacing of the interdigitated finger electrodes changes along the transducer axis.
  • the resonance condition that the frequency is equal to the quotient of the surface sound velocity and the finger spacing is only met in a laterally spatially limited area.
  • an interdigital transducer can also be used as the surface sound wave detector.
  • the analysis method according to the invention can be carried out on a chip on which the probe molecules are directly bound.
  • the liquid drop with the macromolecules to be examined is moved on a preferred location area of the solid surface, the wetting properties of which differ from its surroundings in such a way that the liquid drop is preferably on it.
  • the probe molecules are then arranged on this preferred location area.
  • the area on the solid body on which the liquid is to move can be determined by defining such a preferred location area.
  • the liquid with the macromolecules to be examined is usually only in the preferred location area, without the need for channels or trenches which would otherwise restrict the movement of the liquid.
  • the surface tension of the liquid drop additionally means that the liquid cannot leave the preferred area of residence without the action of an external force.
  • the different wetting properties can e.g. B. can be realized by an appropriate coating either of the preferred lounge area or its surroundings.
  • hydrophilic or hydrophobic areas can be defined.
  • the preferred location is chosen so that it is more hydrophilic than the surrounding solid surface. This can be achieved either by a hydrophilic coating of the preferred area or by a hydrophobic environment.
  • a hydrophobic environment can e.g. B. can be realized by a silanized surface.
  • the solid surface surrounding the area of residence can also be selected to be hydrophilic, lipophobic or lipophilic in comparison to the surface of the preferred area of residence.
  • the wetting properties can also be modulated by microstructuring, as is the case with the so-called lotus effect, which is based on different roughness of the surface and so causes different wetting properties.
  • Such roughness modulation can e.g. B. be obtained by microstructuring the corresponding surface areas, for. B. by chemical treatment or ion radiation.
  • the production of areas with different wetting properties is simple and inexpensive by using already known lithographic processes and / or coating technologies.
  • the geometry of the preferred lounge area can be adapted to the corresponding application.
  • the preferred location area can be meandered through the microarray, so that the individual positions of the microarray line up along the preferred location area.
  • a crosswise arrangement can be provided, in which the individual positions of the microarray are located in the intersection areas.
  • the method according to the invention is carried out on a preferred location area designed in this way, the requirements for the accuracy in the direction and lateral spread of the surface sound wave are lower, since the movement of the liquid drop is also determined by the shape of the location area.
  • correspondingly defined preferred lounge areas are provided only in the area of the individual spots, without being connected to one another.
  • the location areas then serve to localize the drop of liquid in the area of a spot or a group of fewer spots.
  • the stay areas defined in this way act like an “anchor", which makes it easier to locate the drop of liquid at a spot or at a group of fewer spots.
  • the inventive method can, for. B. can be used advantageously for the analysis of oligonucleotides.
  • it is particularly suitable for DNA screening, with the oligonucleotides to be examined e.g. B. consist of short strands of DNA (deoxyribonucleic acid strands).
  • the oligonucleotides to be examined e.g. B. consist of short strands of DNA (deoxyribonucleic acid strands).
  • different macromolecules with specific binding such as different proteins, different antigens or different antibodies can also be provided or analyzed in the microarray.
  • At least one drop of liquid with at least one type of macromolecule is applied to a solid surface, preferably the surface of a chip, the macromolecules having an active group which is activated in this way when a corresponding external parameter changes that it can bond with the surface of the solid or with macromolecules already bound to the surface.
  • the drop is moved to a desired location on the surface with the aid of the impulse transmission of one or more surface sound waves.
  • the external parameter is then changed homogeneously and not locally in such a way that binding of the macromolecules is made possible.
  • the liquid drop can then be moved several times to a different location of the chip with the aid of the surface sound waves and the parameter changed again in order to successively generate a microarray.
  • the lighting is chosen as an external parameter. Depending on the nature of the photoactive group, either its intensity or its frequency is changed in order to enable the photoactive macromolecules to bind to the surface or to macromolecules that have already been bound.
  • the drop of liquid is moved to the desired location using the surface sound wave and then the lighting is switched on or changed accordingly. Only where the liquid is located will the macromolecules bind to the surface. In this way, a spot of a microarray can be generated very precisely and locally.
  • the drop can also contain macromolecules that respond to changes in temperature. If the drop has been moved to the desired location with the aid of surface acoustic waves, the chip surface is heated in order to enable the reaction of the macromolecules with the surface or already bound macromolecules at this location. The temperature increase need not be local, since a spatial resolution is guaranteed by the limited extent of the drop.
  • interdigital transducers e.g. B. so-called tapered interdigital transducers can be used.
  • tapered interdigital transducers can be used.
  • the use of such interdigital transducers in the production method according to the invention offers the same advantages as in the analysis methods according to the invention.
  • the production method according to the invention can take place on the free surface of a solid-state chip.
  • the solid-state chip is provided with a preferred area of residence before use of the production method according to the invention, the wetting properties of which differ from its surroundings in such a way that the liquid with the macromolecules to be bound is preferably thereon.
  • the requirement for the accuracy of the surface sound wave radiation can be reduced, since the movement of the liquid is mainly limited to the preferred location area.
  • an analysis device manufactured with such a production method can be used advantageously to carry out an analysis method according to the invention, in which a preferred lounge area, as already described above, can also be used advantageously.
  • a preferred lounge area can be created as already described in the analysis methods according to the invention. Different geometries of the lounge area are possible.
  • several preferred location areas can also be used, which are arranged at those locations on the surface of the solid body where the spots of the microarray that are produced with the production method according to the invention are to be located. The preferred locations at these locations facilitate the localization of the liquid with the macromolecules to be bound to the spots. By irradiating a surface sound wave of sufficiently high intensity, the liquid drop with the macromolecules which are to form the spots of the microarray can nevertheless be moved from one preferred location area to the next.
  • the amount of macromolecules bound at one location is determined by measuring the attenuation and / or change in speed of at least one surface sound wave which is sent in the direction of the bound macromolecules.
  • a surface sound wave is sent in the direction of the spot or spots and after passing through the corresponding area of the surface, its intensity and / or the speed change is measured.
  • the attenuation or change in speed of the surface sound wave can be determined by the presence of the macromolecules. The amount of bound macromolecules can be deduced from this.
  • the number of macromolecules located there can thus be inferred for each individual spot of the microarray produced. If such a measured microarray for the analysis of macromolecules z. B. used according to the analysis method according to the invention, an accurate quantitative statement is possible. If necessary, a calibration measurement is also carried out.
  • a microarray can be produced in a precise and reproducible manner, which can be used particularly advantageously with the analysis method according to the invention.
  • the devices for generating the surface sound waves preferably the interdigital transducer, can first be used for the production of the microarray and then used to carry out analyzes. Any preferred stay area that may be present can also be advantageously used initially in the manufacture of the microarray and then used in the following analyzes.
  • a small amount of liquid is moved with the aid of surface acoustic waves in order to be able to react locally, either to build up a microarray from individual spots or to react the material in the liquid with individual spots of the microarray.
  • the use of the impulse transmission of surface sound waves enables a gentle and defined movement.
  • FIG. 1 shows an arrangement for carrying out the method according to the invention for analyzing macromolecules
  • FIG. 2 shows another arrangement for carrying out a method according to the invention for analyzing macromolecules
  • FIG. 3 shows a further arrangement for carrying out a method according to the invention for the analysis of macromolecules
  • Figure 4 shows a step in the manufacture of an analysis device according to the invention.
  • Figure 1 shows a device for performing the analysis method according to the invention. 1 identifies the surface of a solid-state chip, similar to how it z. B. is also used for applications in semiconductor technology.
  • the material of the substrate of this surface is piezoelectrically and advantageously coated with silicon dioxide.
  • a silicon dioxide coating guarantees a well-defined and well-known surface chemistry.
  • Interdigital transducers 5, 6, 7 and 8 are in a tapered version on the surface.
  • the interdigital transducers consist of electrodes 9 with finger-like extensions which engage in pairs.
  • other geometries of interdigital transducers are also conceivable, as are known from the technology of surface wave filters.
  • a surface sound wave is generated with a wavelength that corresponds to the finger spacing of the electrodes and whose direction of propagation is essentially perpendicular to the finger electrodes. Due to the tapered design, in which the distance between the fingers differs locally, a laterally limited surface sound wave can be generated.
  • the transducers each include a large number of interlocking fingers, only a few of which are shown schematically and not to scale.
  • the number and arrangement of the interdigital transducers is adapted to the desired circumstances.
  • a preferred lounge area 11 between the interdigital transducers. Its wetting properties differ from its surroundings in such a way that an aqueous solution is preferably located thereon.
  • an aqueous solution is preferably located thereon.
  • e.g. B. silanized the surroundings of the preferred lounge area.
  • Such a preferred lounge area improves the localization of aqueous solution, but is not necessarily provided for carrying out the method according to the invention.
  • Spots 16 made of macromolecules are arranged in the area of the preferred stay area 11. The individual spots 16 each comprise macromolecules, which for each spot 16 z. B. are different.
  • the analysis method according to the invention is carried out as follows.
  • a drop of liquid 3 with macromolecules to be examined is z. B. brought into the area of the spots 16 with the aid of a pipetting robot, as can be seen in FIG. 1.
  • the liquid drop is of the order of a few nl to ⁇ l.
  • a spot has B. an expansion of 100 microns when a drop with a few nl is used.
  • a pulse is generated on the amount of liquid.
  • an electromagnetic alternating field of a few 10 to a few 100 MHz is applied to the corresponding interdigital transducer via the electrodes 9 and corresponding connecting electrodes, not shown.
  • a surface acoustic wave is generated which propagates essentially perpendicular to the fingers of the interdigital transducer. This is indicated by the arrow 14 in FIG. 1 as an example.
  • the surface sound wave transmits an impulse to the liquid quantity 3 and thus moves the liquid drop from one spot to another.
  • the liquid drop 3 can thus be brought to the place of a specific spot 16 in a defined manner.
  • the liquid drop 3 After a residence time, which corresponds approximately to the typical hybridization time of the macromolecules present in the liquid drop 3 with the macromolecules present on the spot, the liquid drop 3 is again removed from the spot with the aid of surface acoustic waves. It can be moved to another spot to enable a corresponding hybridization at the location there. After the desired spots 16 have come into contact with the liquid 3, it is analyzed at which spots a hybridization with the macromolecules present in the liquid has taken place. Are z. B. fluorescence-marked macromolecules in the liquid can be determined by a spatially resolved measurement of the fluorescence, at which spots a reaction has taken place. Is the type of macromolecule that is attached the individual spots 16, known, can be inferred in this way from the type of macromolecules in the liquid drop 3.
  • An in situ measurement can also be carried out.
  • the liquid drop 3 After the liquid drop 3 has been applied to the first spot, it is z. B. with the help of the surface sound wave 14 of the first interdigital transducer 7 moved a little away from the spot.
  • the opposite interdigital transducer 6 With the opposite interdigital transducer 6, a second surface acoustic wave is generated, which acts in alternation with the first surface acoustic wave 14 in the opposite direction on the drop 3.
  • the drop 3 moves backwards over the spot. Due to the temporal change of the two surface sound waves, the drop moves back and forth.
  • the drop 3 touches the spot, macromolecules from the liquid react with macromolecules at the spot.
  • the time course of the reaction can be determined and a statement can be made about the reaction kinetics.
  • the measurement of fluorescence or another detection parameter, such as. B. the electrical charge or the mass, is carried out in a known manner with appropriate measuring and evaluation devices and evaluation programs that are not of interest here.
  • the concentration of the macromolecules to be examined in the liquid drop can be higher, even if only a small amount of starting material is present on the macromolecules to be examined.
  • the concentration is comparable to that of conventional methods, the amount of sample can be much smaller since only nanoliter amounts are required.
  • FIG. 2 shows the geometry of another arrangement for carrying out the analysis method according to the invention.
  • the spots 16 are arranged here along a meandering area 17.
  • a drop of liquid which is located on this meandering location area 17 can be moved along the location area with the aid of the interdigital transducers, so that it touches all spots sequentially.
  • This arrangement of the location area lowers the requirements for the accuracy of the lateral resolution of a single interdigital transducer.
  • a surface sound wave 12 is generated with the aid of the interdigital transducer 8, so that a drop of liquid can be moved upward in the part of the area 17 on the left in the figure. For the sake of clarity, the liquid drop itself is not shown.
  • interdigital transducers Appropriate timing of the interdigital transducers also enables the use of interdigital transducers with an even finger spacing.
  • another interdigital transducer is used synchronously, which enables movement in the bent direction.
  • the evaluation is carried out in the same way as for an arrangement in FIG. 1.
  • the method according to the invention can also be carried out with an analysis device according to FIG. 3.
  • the lounge area 19 is cruciform. Even with such an arrangement, the requirements for the precision of the interdigital transducers are lower.
  • preferred areas of residence are provided only in the area of the individual spots, without being connected to one another. Such location areas serve to localize the drop of liquid in the area of the individual spots. By irradiating a surface sound wave of sufficient intensity, the liquid drop can nevertheless be moved from one area to the next. However, the requirements for the precision of the surface wave radiation are reduced, since the preferred areas of residence at the individual spots support the localization of the drop in their area.
  • the method according to the invention can also be carried out without the use of a preferred location area 11, 17, 19 if the control of the interdigital transducers is precise enough.
  • a number of different and individually controllable transducers can also be used, which always only cover a narrow area of the surface with a surface sound wave.
  • FIG. 4 shows a step in the production method for a microarray according to the invention.
  • a preferred residence area 11 is provided on the solid-state chip 1.
  • a drop of liquid 2 is z. B. applied with the aid of a pipetting robot.
  • the amount of liquid is e.g. B. in the order of 1 nl to a few ⁇ l.
  • the liquid drop contains macromolecules which are intended to serve as probe molecules and are to be bound at predetermined locations for this purpose.
  • the liquid drop is brought to a desired location with the aid of surface sound waves, which are generated with the interdigital transducers 5, 6, 7 and 8.
  • a surface wave 13 is shown as an example, which is generated with the interdigital transducer 7 in a manner which has already been described above for the analysis method according to the invention. is written.
  • the impulse transmission of the surface sound wave 13 to the liquid drop 2 moves it in the direction 15.
  • the sonication is stopped with the surface sound wave, so that the liquid drop 2 comes to rest.
  • an external parameter is changed, to which the material in the liquid drop 2 reacts.
  • the macromolecules contained in the liquid drop can be photoactive.
  • an illumination source of suitable wavelength and intensity is switched on, which is arranged above the chip 1 and illuminates the entire chip homogeneously.
  • the surface chemistry of the chip or the preferred location area 11 is selected such that the macromolecules are bound by the illumination of the photoactive group of the macromolecules to the surface. In this way, a spot is generated at a defined location on the surface, at which the macromolecules from the amount of liquid 2 bind. The amount of liquid 2 can then be moved on to generate another spot at another location.
  • a further amount of liquid can be applied in which there are other macromolecules which are to be bound at other locations on the chip, in order to produce a microarray in which different macromolecules are located at different spots.
  • Liquid drops with different macromolecules can also be moved successively over the same spot. In this way, different sequences can be attached to one another in order to generate desired probe molecules.
  • macromolecules which have an electroactive group which binds to the surface when an electric field is applied, so instead of lighting, a homogeneous electric field of suitable strength, orientation and polarity is created.
  • the entire process can also be parallelized to increase the process speed when manufacturing a microarray.
  • FIGS. 2 and 3 can also be generated in an analogous manner.
  • the location of the liquid droplets that are sent over the surface to form the microarray can be supported by the corresponding preferred location areas. It is thus also possible to provide an arrangement of preferred areas of residence on the surface of the solid body at which the spots are to be located according to the manufacturing process. Such localization areas, which are not connected to one another, support the localization of the liquid in the formation of the microarray, so that requirements for the precision of the movement induced by the surface sound waves are reduced.
  • the external parameter in the examples described the lighting or the electrical field, does not have to be generated locally, but can be homogeneous over the entire chip surface.
  • the spatially resolved binding is ensured by the fact that only a very small part of the surface is connected to the liquid drop 2 in which the macromolecules to be bound are located.

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Abstract

La présente invention concerne un procédé pour analyser des macromolécules à l'aide d'un microréseau constitué de plusieurs taches situées selon une disposition connue. Ces taches comprennent une pluralité de premières macromolécules au moins partiellement différentes. Ledit microréseau se trouve sur une surface solide, de préférence sur une puce. Selon ce procédé, au moins une goutte de liquide présentant des macromolécules est appliquée sur ladite surface solide. La quantité de liquide est si faible que seulement une tache ou un groupe de quelques taches du microréseau vient en contact avec la goutte de liquide. Après un intervalle de temps prédéfini, la goutte de liquide est déplacée vers une tache suivante ou vers un autre groupe de quelques taches, à l'aide d'un report d'impulsion d'une onde sonore de surface. On étudie ensuite si une réaction avec la matière de la goutte de liquide a eu lieu dans les taches qui sont venues en contact avec la goutte de liquide. La présente invention concerne également un procédé pour produire un dispositif d'analyse pour macromolécules. Selon ce procédé, au moins une goutte de liquide présentant au moins un type de macromolécule est appliquée sur une surface solide, de préférence sur une puce. Ces macromolécules présentent un groupe actif qui est activé lors de la modification d'un paramètre extérieur correspondant, afin de permettre une liaison à la surface ou à des molécules déjà liées. La goutte est déplacée à un endroit souhaité sur la surface, à l'aide du report d'impulsion d'une ou de plusieurs ondes sonores de surface. Le paramètre extérieur est modifié de manière homogène et non locale de façon à permettre une liaison des macromolécules, formant la tache d'un microréseau dans laquelle des macromolécules de la goutte de liquide sont liées à la surface. En outre, cette invention concerne un dispositif d'analyse permettant de mettre en oeuvre le procédé d'analyse de la présente invention.
PCT/EP2002/007644 2001-07-24 2002-07-09 Procede pour analyser des macromolecules, dispositif d'analyse et procede pour produire un dispositif d'analyse WO2003012389A2 (fr)

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EP02764659A EP1412533A2 (fr) 2001-07-24 2002-07-09 Procede pour analyser des macromolecules, dispositif d'analyse et procede pour produire un dispositif d'analyse

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DE10136008A DE10136008B4 (de) 2001-07-24 2001-07-24 Verfahren zur Analyse von Makromolekülen und Verfahren zur Herstellung einer Analysevorrichtung

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Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE102004037348A1 (de) * 2004-08-02 2006-03-16 Infineon Technologies Ag Fluid-Transport-Vorrichtung, Sensor-Anordnung, Fluid-Misch-Vorrichtung und Verfahren zum Herstellen einer Fluid-Transport-Vorrichtung
WO2007059839A1 (fr) * 2005-11-28 2007-05-31 Advalytix Ag Procede, dispositif et trousse pour l'analyse de macromolecules dans un echantillon
WO2008064730A2 (fr) * 2006-11-30 2008-06-05 Olympus Life Science Research Europa Gmbh Procédé pour effectuer une réaction enzymatique
WO2008080531A2 (fr) * 2007-01-05 2008-07-10 Advalytix Ag Procédé, dispositif et kit pour l'examen d'un échantillon de liquide
US20110244598A1 (en) * 2008-12-03 2011-10-06 Roche Diagnostics Operations, Inc. Test Element Having Combined Control and Calibration Zone
US8062904B2 (en) 2005-01-05 2011-11-22 Beckman Coulter, Inc. Method and device for dosing and mixing small amounts of liquid
US8186869B2 (en) 2005-01-05 2012-05-29 Beckman Coulter, Inc. Method and device for dosing and mixing small amounts of liquid
EP2809428A4 (fr) * 2012-01-31 2015-11-04 Penn State Res Found Manipulation microfluidique et tri de particules à l'aide d'une onde acoustique de surface stationnaire et synchronisable

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE10328624A1 (de) * 2003-06-25 2005-01-20 Advalytix Ag Verfahren zur Hybridisierung
DE102004051394B4 (de) * 2004-10-21 2006-08-17 Advalytix Ag Verfahren zur Bewegung von kleinen Flüssigkeitsmengen in Mikrokanälen und Mikrokanalsystem

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1994027719A1 (fr) * 1993-05-27 1994-12-08 Protogene Laboratories, Inc. Procede et appareil pour realiser une serie de reactions chimiques sur une surface de support
US5674742A (en) * 1992-08-31 1997-10-07 The Regents Of The University Of California Microfabricated reactor
US6010316A (en) * 1996-01-16 2000-01-04 The Board Of Trustees Of The Leland Stanford Junior University Acoustic micropump
DE19836110A1 (de) * 1998-08-10 2000-02-24 Biotul Bio Instr Gmbh Vorrichtung und Verfahren zur grenzflächennahen Mischung von Proben in Biosensorsystemen

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5846708A (en) * 1991-11-19 1998-12-08 Massachusetts Institiute Of Technology Optical and electrical methods and apparatus for molecule detection
JPH0927590A (ja) * 1995-07-10 1997-01-28 Nippon Inter Electronics Corp 複合半導体装置
DE19827900A1 (de) * 1997-10-21 1999-04-22 Roche Diagnostics Gmbh Verfahren zur Beschichtung von Oberflächen
DE59915204D1 (de) * 1998-08-28 2010-10-28 Febit Holding Gmbh Verfahren zur herstellung von biochemischen reaktionsträgern
FR2783179B1 (fr) * 1998-09-16 2000-10-06 Commissariat Energie Atomique Dispositif d'analyse chimique ou biologique comprenant une pluralite de sites d'analyse sur un support, et son procede de fabrication

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5674742A (en) * 1992-08-31 1997-10-07 The Regents Of The University Of California Microfabricated reactor
WO1994027719A1 (fr) * 1993-05-27 1994-12-08 Protogene Laboratories, Inc. Procede et appareil pour realiser une serie de reactions chimiques sur une surface de support
US6010316A (en) * 1996-01-16 2000-01-04 The Board Of Trustees Of The Leland Stanford Junior University Acoustic micropump
DE19836110A1 (de) * 1998-08-10 2000-02-24 Biotul Bio Instr Gmbh Vorrichtung und Verfahren zur grenzflächennahen Mischung von Proben in Biosensorsystemen

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
WANG J: "SURVEY AND SUMMARY FROM DNA BIOSENSORS TO GENE CHIPS" NUCLEIC ACIDS RESEARCH, OXFORD UNIVERSITY PRESS, SURREY, GB, Bd. 28, Nr. 16, 2000, Seiten 3011-3016, XP001127052 ISSN: 0305-1048 *
WIXFORTH A., SCRIBA J.: "Nanopumpen für das Chiplabor" GIT LABOR-FACHZEITSCHRIFT, Bd. 05, 2002, Seite 594 XP009009065 *

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE102004037348A1 (de) * 2004-08-02 2006-03-16 Infineon Technologies Ag Fluid-Transport-Vorrichtung, Sensor-Anordnung, Fluid-Misch-Vorrichtung und Verfahren zum Herstellen einer Fluid-Transport-Vorrichtung
US8062904B2 (en) 2005-01-05 2011-11-22 Beckman Coulter, Inc. Method and device for dosing and mixing small amounts of liquid
US8186869B2 (en) 2005-01-05 2012-05-29 Beckman Coulter, Inc. Method and device for dosing and mixing small amounts of liquid
WO2007059839A1 (fr) * 2005-11-28 2007-05-31 Advalytix Ag Procede, dispositif et trousse pour l'analyse de macromolecules dans un echantillon
WO2008064730A2 (fr) * 2006-11-30 2008-06-05 Olympus Life Science Research Europa Gmbh Procédé pour effectuer une réaction enzymatique
WO2008064730A3 (fr) * 2006-11-30 2008-07-24 Advalytix Ag Procédé pour effectuer une réaction enzymatique
WO2008080531A2 (fr) * 2007-01-05 2008-07-10 Advalytix Ag Procédé, dispositif et kit pour l'examen d'un échantillon de liquide
WO2008080531A3 (fr) * 2007-01-05 2008-12-31 Advalytix Ag Procédé, dispositif et kit pour l'examen d'un échantillon de liquide
US20110244598A1 (en) * 2008-12-03 2011-10-06 Roche Diagnostics Operations, Inc. Test Element Having Combined Control and Calibration Zone
US8569073B2 (en) * 2008-12-03 2013-10-29 Roche Diagnostics Operations Inc. Test element having combined control and calibration zone
EP2809428A4 (fr) * 2012-01-31 2015-11-04 Penn State Res Found Manipulation microfluidique et tri de particules à l'aide d'une onde acoustique de surface stationnaire et synchronisable

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WO2003012389A3 (fr) 2003-07-17

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