EP2004317A1 - Apparatus for enhancing the reaction efficiency, especially the binding efficiency, between molecules and molecular moieties - Google Patents

Apparatus for enhancing the reaction efficiency, especially the binding efficiency, between molecules and molecular moieties

Info

Publication number
EP2004317A1
EP2004317A1 EP07718377A EP07718377A EP2004317A1 EP 2004317 A1 EP2004317 A1 EP 2004317A1 EP 07718377 A EP07718377 A EP 07718377A EP 07718377 A EP07718377 A EP 07718377A EP 2004317 A1 EP2004317 A1 EP 2004317A1
Authority
EP
European Patent Office
Prior art keywords
micro
reaction
array
magnetic
magnetic coil
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.)
Granted
Application number
EP07718377A
Other languages
German (de)
French (fr)
Other versions
EP2004317B1 (en
Inventor
Rudolf Heer
Christa NÖHAMMER
Jörg SCHOTTER
Moritz Eggeling
Rudolf Pichler
Markus Mansfeld
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.)
AIT Austrian Institute of Technology GmbH
Tecnet Equity NO Technologiebeteiligungs Invest GmbH
Original Assignee
Tecnet Capital Technologiemanagement GmbH
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 Tecnet Capital Technologiemanagement GmbH filed Critical Tecnet Capital Technologiemanagement GmbH
Publication of EP2004317A1 publication Critical patent/EP2004317A1/en
Application granted granted Critical
Publication of EP2004317B1 publication Critical patent/EP2004317B1/en
Not-in-force legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F33/00—Other mixers; Mixing plants; Combinations of mixers
    • B01F33/45—Magnetic mixers; Mixers with magnetically driven stirrers
    • B01F33/451—Magnetic mixers; Mixers with magnetically driven stirrers wherein the mixture is directly exposed to an electromagnetic field without use of a stirrer, e.g. for material comprising ferromagnetic particles or for molten metal
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F33/00—Other mixers; Mixing plants; Combinations of mixers
    • B01F33/30—Micromixers
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip

Definitions

  • the invention is concerned with the targeted increase in the hit or hit probability between reactive partners capable of binding together, which on the one hand increases the amount of binding products and, on the other hand, significantly shortens the reaction time.
  • bioanalyzer arrays such as in e.g. is shortened in DNA / RNA, protein or immuno-microarrays, thereby simultaneously achievable increase in the evaluable fluorescence signals.
  • This is achieved by agitation of the hybridization solution using magnetic micro- or nanoparticles, which are moved in a controlled manner by the hybridization solution and transport the target molecules to the individual spots, ie binding sites.
  • the range of action of the individual spots which is diffusion-limited in conventional analytical methods, is thus substantially increased or expanded.
  • the invention has the further advantage that it can be combined in a simple manner with already existing bioanalytical array systems, so that it makes it possible to retrofit already existing reaction systems, thus in particular already existing bioanalysis systems.
  • DNA microarrays represent a real technological leap in the detection of genes and gene defects, in gene expression analysis, and in the understanding of gene functions.
  • a microbio array or biochip typically consists of a specially chemically coated glass slide containing up to several thousand microscopically differently functionalized spots, known as spots.
  • spots In the case of DNA microarrays or DNA chips, each of these spots consists of numerous copies of a distinct DNA segment or gene, which serve as "capture molecules" or probes for corresponding specific DNA or mRNA present in a sample to be analyzed.
  • the target molecules are previously labeled with fluorescent particles in order to be able to detect them after binding to the corresponding chip probes by means of fluorescence scanners.
  • the factors specificity and sensitivity play in the biological or
  • Microarray analysis plays a big role. While the specificity is essentially of the
  • Hybridization buffer and the reaction temperature the sensitivity depends on
  • Fluorescence labeling of the targets to some extent also on the amount of DNA present on the chip and the efficiency of binding between the target and probe.
  • SlideBooster Surface acoustic waves or surface acoustic waves are used in an already commercially available product, called SlideBooster from Advalytix AG, Eugen-Shacker-Strasse 53.0, D-85649 Brunnthal, Germany, www.advalytix.de, SlideBooster SB400, for the mixing of thin liquid films used.
  • Bubble-induced acoustic micromixing Lab Chip, 2002, 2, 151-157. To realize the mixing effect are induced by bubbles, acoustic
  • Microstreams used which are generated by means of a piezoelectric sounder.
  • To increase the efficiency in the mixing chamber microscopic, mechanically manufactured, pockets installed, where gas bubbles form.
  • This modification of the mixing chamber represents a significant overhead in the production of bio-chips and is questionable in terms of reusability due to contamination.
  • a mixing device for bio-sensors in which by means of a stirrer by a mechanical shaft from the outside in a chamber, a homogenization of the sample is effected, described in WO 94/28396. In this case, the stirrer performs a mutual movement normal to the surface of the sensor during the measurement of the signals.
  • Another patent, namely GB 876 070 describes in general the
  • the mixing of thin liquid layers which comprise a suspension of mobile magnetic particles is described in EP 0 240 862 A1.
  • the device also includes magnet systems. The arrangement described there provides a gap for receiving the liquid film with the permanent magnetic particles.
  • WO 97/02357 A1 contemplates a mixing device for use with DNA chips. There are mentioned an acoustic mixture and a magnetic mixture, which is caused by alternating currents in electromagnets.
  • US Pat. No. 6,806,050 B2 discloses an electromagnetic chip or biochip which comprises a matrix of individually-susceptible microelectromagnet units and on whose surface special molecules are immobilized. By means of the magnet units, molecules bound to small magnetic particles are moved essentially in the plane of the biochip, and in this way an increase in the number of bonds is achieved.
  • the newly developed device according to the invention is based on increasing the range of action of the individual binding sites, for example DNA spots, by adding magnetic micro- or nanoparticles to the hybridization solution, which are moved by means of an externally generated magnetic field and, for example, the DNA targets the probes of the individual spots are much more targeted approachable. It be carried by the or by means of the magnetic particles, the DNA targets in a microflow and transported in this way.
  • the present invention is an apparatus for increasing the frequency or frequency of contact between two reactants capable of binding together, preferably between the analyzer molecule or molecular moiety and the analyte molecule or moiety, in particular to increase the binding efficiency and to reduce the binding efficiency Binding times required in a bioanalysis arrays according to the terms of claim 1, which have the features mentioned in the characterizing part of this claim.
  • the combination of the matrix-like arrangement of the micro or milli-magnetic coils with only one opposing central magnetic coil allows a targeted, precise movement of the magnetic particles within the reaction liquid, ie in particular of the hybridization liquid film and thus in a controlled controlled approaching of target DNA to the immobilized probes on the DNA Bio-Chip.
  • the special arrangement of the micro or Milli coils and the "pattern" for their supply with magnetizing current prevent accumulation of the magnetic particles and thus represents a significant advantage over the previously known, above-based, based on the movement of magnetic particles, mixing devices
  • the use of magnetic fields also allows for easy combination with a sample chamber in which the humidity and temperature are controlled, allowing a high degree of system integration.
  • the single magnetic coil above the chip serves to cause the magnetic particles to move upwards. If this coil is no longer magnetic by switching off the current, the magnetic particles begin to sink again. A drop in the same path as the lift path is prevented by the fact that the micro-magnetic coils of the micro-magnetic matrix below the chip, eg wave-like magnetized and thus the magnetic particles are displaced sideways during their sinking and thus ultimately a kind of oblique flow in the reaction liquid is induced, by which the target molecules more moved and thus more probe molecules are supplied.
  • the particles are each moved laterally in the direction of the center of the correspondingly connected micro-magnetic matrix.
  • the inventively provided arrangement allows in contrast to the aforementioned arrangements according to the prior art, which deal with the non-directional or at most in one direction mixing of liquid films, a preprogrammed, very targeted movement of the micro-magnetic particles used.
  • the device according to the invention in particular the clumping and accumulation of the magnetic particles.
  • the invention has the advantage that it can be easily combined with already established, existing bio-analysis arrays and further, that when integrating the magnetic coils in a corresponding carrier, the temperature at the interface with the bio-chip can be precisely adjusted, which later to be discussed integrated cooling / heating circuit is made possible.
  • the device according to the invention thus differs in particular from an arrangement which exclusively aims at possible process steps in an integrated microfluidic biochip and is not suitable for retrofitting already existing DNA microarrays.
  • a driving device for the micro-magnetic coils on the one hand and the single solenoid on the other hand according to A nspruch 2 is provided by means of which each individual micro-solenoid is individually controlled and by means of which each time-dependent magnetization pattern on the magnet Matrix surface can be imprinted.
  • optical control is a
  • the invention relates to a specific arrangement of the components of the device according to the invention.
  • the invention relates to a preferred embodiment of the device with a receiving chamber for the reaction vessel, in particular for the bio-analysis array microchip.
  • FIG. 1 shows the basic arrangement of the essential components of the device according to the invention
  • FIG. 2 shows schematically the processes during
  • FIG. 3 shows schematically the control of the new device
  • Fig. 4 shows an example of a real embodiment of the new device
  • Fig. 5 shows schematically a diagram of the path which a magnetic particle travels within the sample liquid
  • Fig 6a shows a diagram which shows the signal increase when using the new device in comparison
  • Fig. 6b shows a diagram in which the signal increase as a function of the hybridization time is also shown in comparison with the prior art:
  • Fig. 1 In the oblique view of Fig. 1 is the basic structure of the device 1 according to the invention for a magnet-induced intensification of the contacts between two reaction, in particular binding partners, such. Target molecule and probe molecule shown.
  • On the glass slide 4 is below the coverslip 5 of the
  • Bioanalysis array 6 with arranged in regularly applied fine spots with the probe molecules or the bio-chip 6. Between slide 4 and coverslip 5 is the reaction, in particular hybridization liquid with the target molecules and provided for the agitation of the liquid micro-magnetic particles.
  • the here flat ring-like or toriodförmige single magnetic coil 3 without core so with a free central opening 36th arranged, through which the review of the reaction on the slide 4 and below the coverslip 5 is kept free.
  • micro-magnet coils 2 with coil cores 21, preferably in a hexagonal matrix 20, can be arranged underneath the microscope slide 4.
  • FIG. 2 shows - with otherwise constant reference numerals - as between the slide 4 with the spots with the probe molecules 63 of the bio-chip 6, the reaction liquid 70 is located, and as in the same - from the previously described electro Magnets 2 and 20, 3 and their variable magnetic fields acted upon, the magnetic particles 75 shown here circular move approximately in the symbolized by arrows movement directions and the symbolized by asterisks - fluorescent dye 72 acted upon target molecules 73 with the immobilized probe molecules 63 in contact bring and bind there much more often than this, for example would be achievable by diffusion.
  • the control device 8 shown very schematically in FIG. 3 - with otherwise identical reference numerals - is used for individual energization of the micro-magnetic coils 2 of the micro-magnetic matrix 20 with variable magnetizing currents as well as for the likewise variable supply of the individual magnetic coil 3 with current.
  • the central control unit (PC control) 81 is connected to a control unit, eg D / A card 82, which is connected on the one hand to the power supply unit (power supply 1) 83 for the micro-solenoid matrix 20 and on the other hand to the power supply unit (FIG. Power Supply 2) 84 for the single solenoid 3 is connected.
  • the power supply unit (power supply 1) 83 is connected to a - connected directly to the D / A card 82 relay matrix unit 85, from which the time-dependent variable individual power supply of each individual connected to the same single micro-magnetic coil 2 of the magnetic matrix 20 according to a is performed by the central control unit 81 program. It is also planned to control the temperature of the sample.
  • thermocontrol temperature control unit
  • a temperature control unit thermocontrol
  • thermosensors not shown here, which are arranged in the sample area or in the vicinity of the magnets 2, 20 and 3
  • Fig. 4 shows - with otherwise identical reference numerals - in each case in sectional views from the side and from above, the real configuration of the components in the new improved analysis device 1.
  • cover unit II and a Basic block unit I shown, between which the slide not shown here is arranged with the reaction to be carried out.
  • the basic block unit I consists - as can be seen from FIG.
  • Matrix 20 is another small, also from a cooling or Schumediumskanal 225 traversed aluminum block 25 is arranged.
  • the aluminum block 21 Covered at the top is the aluminum block 21 with a thin film 23, on which peripherally surrounding rubber rings 26,27 rest, within which there is a space 230 for the placement of the sample.
  • the cover unit II shown in Fig. 4c comprises a - here with the center a recess 34 having stainless steel top surface 35 - limited upwards aluminum block 31, which is also crossed by cooling / Schumediumskanälen 32 and in which the annular single solenoid 3 is housed ,
  • the aluminum block 31 is penetrated by a central opening 34 ', through which the view of the sample is kept free, and closed at the bottom with a glass plate 33.
  • thermocouples 300 which deliver the current temperature data to the above-mentioned thermal control unit.
  • FIG. 4d provides an overview of the course of the
  • Fig. 5 shows - with otherwise constant reference numerals - schematically the path or course of motion of the micro-magnetic particles 75 in
  • Magnetic particle 75 deflected laterally and then follows about the web B, etc.
  • FIGS. 6a and 6b show the signal increases achieved with the device according to the invention as a function of the concentration of magnetic particles after hybridization: On the ordinate of the diagram Fig. 6a is the intensity of the fluorescence indication and on the abscissa is the concentration of the magnetic particles M-PVA 13 bead (5-8 ⁇ m) (Chemagen AG, Arnold-Sommerfeld-Ring 2, D-52499 Baesweiler) in ⁇ g / ⁇ l.
  • Mean is shown in the right column as a small red cross. In the middle
  • the intensities I of the fluorescence signals obtained using the device according to the invention are also compared with the intensities of the signals obtained in a previously known manner as a function of the hybridization time, with otherwise identical reference numerals.
  • the hybridization time in minutes is plotted on the abscissa and the concentration of micro-magnetic particles or beads M-PVA 13 bead 5-8 ⁇ m is kept constant at [1.8 ⁇ g / ⁇ l]. It shows up after 5 min, a very large signal gain when using the device according to the invention.
  • SlideBooster SB400 (2) R.H. Liu et al. Bubble-induced acoustic micromixing, Lab Chip, 2002, 2, 151-157

Abstract

An apparatus for enhancing the contact frequency between two reactants capable of binding to one another, preferably between an analyzer molecule and an analyte molecule, especially for enhancing the binding effectiveness in bioanalysis arrays with the aid of micro- or nanomagnetic particles (75) set in motion in a controlled manner in the fluid reaction medium by means of magnetic fields generated by variably feedable electromagnets (3, 2, 20) arranged on both sides of the reaction fluid film. On one side of a reaction fluid film, especially of a microscope slide (4) comprising the reactants (63), the reaction liquid (70) comprising the reactants (73) and preferably a glass plate (5) covering the microbioanalysis array (6), is arranged close to a two-dimensional matrix (20) having a multitude of miniature or millimagnetic coils (2) feedable individually with magnetization current of variable strength and/or voltage as a function of time-corresponding to a time-dependent variable magnetization pattern desired in each case-and, on the other side of the reaction vessel, especially the microscope slide (4) with the (micro)bioanalysis array (6), in whose vicinity is positioned only one magnetic coil (3) likewise feedable with variable magnetization current and whose magnetic field permeates the entire reaction vessel, especially the microbioanalysis array (6).

Description

Vorrichtung zur Steigerung der Reaktions-, insbesondere der Anbindungseffizienz zwischen Molekülen bzw. Molekülteilen Device for increasing the reaction, in particular the binding efficiency between molecules or molecular parts
Die Erfindung befasst sich mit der gezielten Erhöhung der Treffer bzw. Trefferwahrscheinlichkeit zwischen miteinander bindungsfähigen Reaktionspartnern, wodurch einerseits die Menge an Bindungsprodukten erhöht und anderseits die Reaktionszeit wesentlich verkürzt wird.The invention is concerned with the targeted increase in the hit or hit probability between reactive partners capable of binding together, which on the one hand increases the amount of binding products and, on the other hand, significantly shortens the reaction time.
Sie ist insbesondere darauf gerichtet, dass die erforderlichen Hybridisierungszeiten in Bioanalyse-Arrays, wie in z.B. in DNA/RNA-, Protein- oder Immuno-Mikroarrays, bei dadurch gleichzeitig erreichbarer Steigerung der auswertbaren Fluoreszenzsignale verkürzt wird. Dies wird durch Agitation der Hybridisierungs-Lösung mit Hilfe magnetischer Mikro- oder Nanopartikel, welche in kontrollierter Weise durch die Hybridisierungslösung bewegt werden und die Target Moleküle zu den einzelnen Spots, also Bindungsstellen transportieren, erreicht. Der Wirkungsbereich der einzelnen Spots, welcher in herkömmlichen Analyseverfahren diffusions-limitiert ist, wird somit wesentlich vergrößert bzw. erweitert. Die Erfindung hat den weiteren Vorteil, dass sie in einfacher Weise mit schon vorhandenen Bioanalyse-Array-Systemen kombiniert werden kann, dass sie also ein Nachrüsten schon bestehender Reaktionssysteme, also insbesondere schon vorhandener Bioanalyse-Systeme, ermöglicht.It is particularly directed to ensuring that the required hybridization times in bioanalyzer arrays, such as in e.g. is shortened in DNA / RNA, protein or immuno-microarrays, thereby simultaneously achievable increase in the evaluable fluorescence signals. This is achieved by agitation of the hybridization solution using magnetic micro- or nanoparticles, which are moved in a controlled manner by the hybridization solution and transport the target molecules to the individual spots, ie binding sites. The range of action of the individual spots, which is diffusion-limited in conventional analytical methods, is thus substantially increased or expanded. The invention has the further advantage that it can be combined in a simple manner with already existing bioanalytical array systems, so that it makes it possible to retrofit already existing reaction systems, thus in particular already existing bioanalysis systems.
Als neue Werkzeuge der Molekularbiologie stellen beispielsweise DNA- Microarrays einen echten Technologiesprung beim Nachweis von Genen und Gendefekten, bei der Genexpressions-Analyse und bei der Vertiefung des Verständnisses von Genfunktionen dar.As new tools of molecular biology, for example, DNA microarrays represent a real technological leap in the detection of genes and gene defects, in gene expression analysis, and in the understanding of gene functions.
Ein Mikrobio-Array oder Biochip besteht üblicherweise aus einem, in spezieller Weise chemisch beschichteten, Glas-Objektträger, der bis zu einige Tausend mikroskopisch kleine, unterschiedlich funktionalisierte Punkte, so genannte Spots, enthält. Im Falle der DNA-Microarrays oder DNA-Chips besteht jeder einzelne dieser Spots aus zahlreichen Kopien eines eindeutig bestimmten DNA-Abschnitts bzw. Gens, die als „Fängermoleküle" oder Sonden für entsprechende, in einer zu analysierenden Probe vorhandene, spezifische DNA oder mRNA-Moleküle, also Targets, fungieren. Die Targetmoleküle werden zuvor mit Fluoreszenzpartikeln markiert, um sie nach erfolgter Bindung an die entsprechenden Chip-Sonden mittels Fluoreszenzscanner detektieren zu können. Die Faktoren Spezifizität und Sensitivität spielen in der Bio- bzw.A microbio array or biochip typically consists of a specially chemically coated glass slide containing up to several thousand microscopically differently functionalized spots, known as spots. In the case of DNA microarrays or DNA chips, each of these spots consists of numerous copies of a distinct DNA segment or gene, which serve as "capture molecules" or probes for corresponding specific DNA or mRNA present in a sample to be analyzed. The target molecules are previously labeled with fluorescent particles in order to be able to detect them after binding to the corresponding chip probes by means of fluorescence scanners. The factors specificity and sensitivity play in the biological or
Mikroarrayanalyse eine große Rolle. Während die Spezifizität im Wesentlichen von derMicroarray analysis plays a big role. While the specificity is essentially of the
Auswahl bzw. Sequenzabfolge der DNA-Sonden am Chip abhängig ist und jenenSelection or sequence sequence of the DNA probes depends on the chip and those
Bedingungen, unter denen man den Andockprozess bzw. Hybridisierung zwischen Targets und Sonden ablaufen lässt, also z.B. die Salzkonzentration desConditions under which docking or hybridization between targets and probes occurs, e.g. the salt concentration of the
Hybridisierungspuffers und die Reaktionstemperatur, hängt die Sensitivität imHybridization buffer and the reaction temperature, the sensitivity depends on
Wesentlichen von der vorhandenen Menge des jeweiligen Targets, von der Effizienz derSubstantially from the existing quantity of the respective target, from the efficiency of the
Fluoreszenzmarkierung der Targets, bis zu einem gewissen Grad auch von der Menge der am Chip vorhandenen DNA-Sonde sowie von der Effizienz der Bindung zwischen Target und Sonde ab.Fluorescence labeling of the targets, to some extent also on the amount of DNA present on the chip and the efficiency of binding between the target and probe.
Im klassischen Chip-Experiment ist der Transport der Targets zu den Sonden im wässrigen Milieu allein durch die Diffusion gesteuert. In der Praxis wird eine wässrige Pufferlösung mit den fluoreszenz-markierten Targetmolekülen auf den Chip mit den DNA- Sonden aufgebracht und danach wird ein dünnes Glasplättchen bzw. Deckgläschen darüber gelegt. Dadurch wird ein dünner Flüssigkeitsfilm zwischen Chip und Deckgläschen ausgebildet, innerhalb dessen sich die Targetmoleküle durch freie Diffusion bewegen.In the classic chip experiment, the transport of the targets to the probes in an aqueous environment is controlled solely by the diffusion. In practice, an aqueous buffer solution containing the fluorescence-labeled target molecules is applied to the chip with the DNA probes, and then a thin glass slide or cover slip is placed over it. As a result, a thin liquid film is formed between the chip and the coverslip, within which the target molecules move by free diffusion.
Bisher wurden unterschiedliche Versuche zur Steigerung der auswertbaren Signale in DNA-Microarrays mit unterschiedlichen Misch- oder Pumpvorrichtungen unternommen, welche sich in folgende 3 Gruppen einteilen lassen.So far, different attempts have been made to increase the evaluable signals in DNA microarrays with different mixing or pumping devices, which can be divided into the following 3 groups.
Mechanische Mischung durch Schütteln und/oder Rotieren des Chips auf einer dafür vorgesehene EinrichtungMechanical mixing by shaking and / or rotating the chip on a dedicated device
Durchpumpen und Rezirkulieren der Hybridisierungs-Lösung am Chip mittels eines externen Fluidiksystems - Mischung der Flüssigkeit am Chip selbstPumping and recirculating the hybridization solution on the chip by means of an external fluidic system - mixing the liquid on the chip itself
Nachfolgend werden Beispiele zu den genannten, dem Stand der Technik entsprechenden Verfahren beschrieben, um Unterscheidungsmerkmale zur vorliegenden Erfindung herausarbeiten zu können.In the following, examples of the aforementioned prior art methods will be described in order to elaborate distinguishing features of the present invention.
Surface Acoustic Waves bzw. akustische Oberflächenwellen werden in einem bereits kommerziell erhältlichen Produkt, mit der Bezeichnung SlideBooster der Firma Advalytix AG, Eugen-Sänger-Straße 53.0, D-85649 Brunnthal, www.advalytix.de, SlideBooster SB400, zur Durchmischung von dünnen Flüssigkeitsfilmen eingesetzt.Surface acoustic waves or surface acoustic waves are used in an already commercially available product, called SlideBooster from Advalytix AG, Eugen-Sänger-Strasse 53.0, D-85649 Brunnthal, Germany, www.advalytix.de, SlideBooster SB400, for the mixing of thin liquid films used.
Eine aktive Durchmischung von Flüssigkeiten wurde von R. H. Liu R. H. Liu et al.Active mixing of liquids was reported by R.H. Liu R.H. Liu et al.
Bubble-induced acoustic micromixing, Lab Chip, 2002, 2, 151-157 vorgestellt. Zur Realisierung der Mischwirkung werden durch Bläschen induzierte, akustischeBubble-induced acoustic micromixing, Lab Chip, 2002, 2, 151-157. To realize the mixing effect are induced by bubbles, acoustic
MikroStrömungen verwendet, welche mittels eines piezoelektrischen Schallgebers erzeugt werden. Zur Steigerung der Effizienz sind in der Mischungskammer mikroskopisch kleine, mechanisch gefertigte, Taschen eingebaut, an denen sich Gasbläschen bilden. Diese Modifikation der Mischungskammer stellt einen beachtlichen Mehraufwand bei der Fertigung von Bio-Chips dar und ist hinsichtlich Wiederverwendbarkeit aus Gründen der Kontamination fragwürdig. Weiters ist eine Mischvorrichtung für Bio-Sensoren, bei der mittels Rührer durch eine mechanische Welle von außen in einer Kammer eine Homogenisierung der Probe bewirkt wird, in der WO 94/28396 beschrieben. Dabei wird vom Rührer eine wechselseitige Bewegung normal zur Oberfläche des Sensors während der Messung der Signale durchgeführt. Eine weitere Patenschrift, nämlich GB 876 070 beschreibt ganz allgemein dieMicrostreams used, which are generated by means of a piezoelectric sounder. To increase the efficiency in the mixing chamber microscopic, mechanically manufactured, pockets installed, where gas bubbles form. This modification of the mixing chamber represents a significant overhead in the production of bio-chips and is questionable in terms of reusability due to contamination. Furthermore, a mixing device for bio-sensors, in which by means of a stirrer by a mechanical shaft from the outside in a chamber, a homogenization of the sample is effected, described in WO 94/28396. In this case, the stirrer performs a mutual movement normal to the surface of the sensor during the measurement of the signals. Another patent, namely GB 876 070 describes in general the
Durchmischung von Flüssigkeiten mittels rotierender Gitter.Mixing of liquids by means of rotating grids.
Das grenzflächennahe Mischen einer zu untersuchenden Flüssigkeit in Sensor- Vorrichtungen wird in der WO 00/09991 A1 beschrieben. Hier erfolgt das Durchmischen mittels Bewegung von magnetischen Kügelchen oder mittels beweglicher Netze, wobei im ersteren Fall die magnetischen Kügelchen in einer Flüssigkeit abwechselnd zwischen zwei Elektromagneten nach oben und nach unten gezogen werden.The near-surface mixing of a liquid to be examined in sensor devices is described in WO 00/09991 A1. Here, the mixing takes place by means of movement of magnetic beads or by means of movable nets, wherein in the former case, the magnetic beads are alternately pulled in a liquid between two electromagnets up and down.
Speziell die Durchmischung von dünnen Flüssigkeitsschichten, welche eine Suspension beweglicher magnetischer Partikel beinhalten, wird in der EP 0 240 862 A1 beschrieben. Die Vorrichtung umfasst ebenfalls Magnetsysteme. Die dort beschriebene Anordnung stellt einen Spalt für die Aufnahme des Flüssigkeitsfilms mit den permanentmagnetischen Partikeln zur Verfügung.In particular, the mixing of thin liquid layers which comprise a suspension of mobile magnetic particles is described in EP 0 240 862 A1. The device also includes magnet systems. The arrangement described there provides a gap for receiving the liquid film with the permanent magnetic particles.
In einem weiteren Patent, nämlich WO 97/02357 A1 ist eine Mischvorrichtung für den Einsatz bei DNA Chips in Erwägung gezogen. Dort werden eine akustische Mischung und eine magnetische Mischung, welche durch Wechselströme in Elektromagneten bewirkt wird, erwähnt.Another patent, WO 97/02357 A1, contemplates a mixing device for use with DNA chips. There are mentioned an acoustic mixture and a magnetic mixture, which is caused by alternating currents in electromagnets.
Aus der US 6,806,050 B2 ist ein elektromagnetischer Chip bzw. Bio-Chip bekannt, welcher eine Matrix von individuell anspeisbaren Micro-Elektromagneteinheiten umfasst und an dessen Oberfläche Sonder-Moleküle immobilisiert sind. Mittels der Magneteinheiten werden an kleine Magnetpartikel gebundene Moleküle im Wesentlichen in der Ebene des Bio-Chips bewegt und es wird auf diese Weise eine Erhöhung der Zahl der Bindungen erreicht.US Pat. No. 6,806,050 B2 discloses an electromagnetic chip or biochip which comprises a matrix of individually-susceptible microelectromagnet units and on whose surface special molecules are immobilized. By means of the magnet units, molecules bound to small magnetic particles are moved essentially in the plane of the biochip, and in this way an increase in the number of bonds is achieved.
Die neu entwickelte Vorrichtung gemäß der Erfindung beruht auf der Vergrößerung des Wirkungsbereichs der einzelnen Bindungsstellen, also z.B. DNA-Spots, durch Beimengung magnetischer Mikro- oder Nanopartikel zur Hybridisierungs-Lösung, welche mittels eines extern erzeugten Magnetfeldes bewegt werden und wobei z.B. die DNA Targets an die Sonden der einzelnen Spots wesentlich gezielter heranführbar sind. Es werden von den bzw. mittels der magnetischen Partikel die DNA-Targets in einer Mikroströmung mitgeführt und auf diese Weise transportiert.The newly developed device according to the invention is based on increasing the range of action of the individual binding sites, for example DNA spots, by adding magnetic micro- or nanoparticles to the hybridization solution, which are moved by means of an externally generated magnetic field and, for example, the DNA targets the probes of the individual spots are much more targeted approachable. It be carried by the or by means of the magnetic particles, the DNA targets in a microflow and transported in this way.
Gegenstand der vorliegenden Erfindung ist eine Vorrichtung zur Steigerung der Kontaktfrequenz bzw. -häufigkeit zwischen zwei miteinander zur Bindung befähigten Reaktionspartnem, vorzugsweise zwischen Analysator-Molekül bzw. -Molekülteil und Analyt-Molekül bzw. -Molekülteil, insbesondere zur Steigerung der Bindungseffektivität und zur Reduktion der für einen Nachweis erforderlichen Bindungszeiten in Bioanalyse- Arrays gemäß dem O b e r b e g r i f f des A n s p r u c h e s 1, welche die im k e n n z e i c h n e n d e n T e i l dieses Anspruches genannten M e r k m a l e aufweist.The present invention is an apparatus for increasing the frequency or frequency of contact between two reactants capable of binding together, preferably between the analyzer molecule or molecular moiety and the analyte molecule or moiety, in particular to increase the binding efficiency and to reduce the binding efficiency Binding times required in a bioanalysis arrays according to the terms of claim 1, which have the features mentioned in the characterizing part of this claim.
Insbesondere die Kombination der matrixförmigen Anordnung der Mikro- bzw. Milli-Magnetspulen mit nur einer gegenüberliegenden zentralen Magnetspule gestattet eine gezielte, präzise Bewegung der magnetischen Partikel innerhalb der Reaktionsflüssigkeit, also insbesondere des Hybridisierungs-Flüssigkeitsfilms und somit im Konkreten ein kontrolliertes Heranführen von Target-DNA an die immobilisierten Sonden auf dem DNA-Bio-Chip.In particular, the combination of the matrix-like arrangement of the micro or milli-magnetic coils with only one opposing central magnetic coil allows a targeted, precise movement of the magnetic particles within the reaction liquid, ie in particular of the hybridization liquid film and thus in a controlled controlled approaching of target DNA to the immobilized probes on the DNA Bio-Chip.
Die spezielle Anordnung der Mikro- bzw. Milli-Spulen und die "Muster" zu deren Anspeisung mit Magnetisierungsstrom verhindern ein Anhäufen der magnetischen Partikel und stellt somit einen wesentlichen Vorteil gegenüber den bisher bekannten, oben genannten, auf der Bewegung magnetischer Partikel beruhender, Mischvorrichtungen dar. Die Verwendung magnetischer Felder ermöglicht außerdem die einfache Kombination mit einer Probenkammer, in welcher die Feuchte und Temperatur kontrolliert wird, so dass ein hoher Grad an Systemintegration ermöglicht wird.The special arrangement of the micro or Milli coils and the "pattern" for their supply with magnetizing current prevent accumulation of the magnetic particles and thus represents a significant advantage over the previously known, above-based, based on the movement of magnetic particles, mixing devices The use of magnetic fields also allows for easy combination with a sample chamber in which the humidity and temperature are controlled, allowing a high degree of system integration.
Die matrix-artige bzw. array-förmige Anordnung der Milli-Magnetspulen mit oder ohne Magnetkerne(n), z.B. unterhalb des DNA-Chips und nur einer Magnet-Spule oberhalb des DNA-Chips ermöglicht eine ganz gezielte Bewegung der magnetischen Partikel.The matrix-like arrangement of the Milli magnet coils with or without magnetic cores (n), e.g. Below the DNA chip and only one magnetic coil above the DNA chip allows a very targeted movement of the magnetic particles.
So dient die einzelne Magnetspule oberhalb des Chips dazu, die magnetischen Partikel zu einer Aufwärtsbewegung zu veranlassen. Wenn diese Spule durch Abschalten des Stromes nicht mehr magnetisch ist, beginnen die Magnetpartikel wieder abzusinken. Ein Absinken in der gleichen Bahn wie die Anhebungsbahn wird dadurch verhindert, dass die Mikro-Magnetspulen der Mikro-Magnet-Matrix unterhalb des Chips, z.B. wellenartig, magnetisiert werden und somit die Magnetpartikel während ihres Absinkens seitwärts verschoben werden und somit letztlich eine Art Schrägströmung in der Reaktionsflüssigkeit induziert wird, durch welche die Target-Moleküle mehr bewegt und dadurch auch mehr Sonden-Molekülen zugeführt werden. Es werden also mittels des erfindungsgemäß vorgesehenen Aufbaus die Partikel jeweils lateral in Richtung des Zentrums der entsprechend beschalteten Mikro-Magnet- Matrix bewegt. Durch Variation der Dauer und der relativen Stärke der Impulse von jeweils benachbarten Mikro-Magnetspulen und der oberhalb des Chips angeordneten Magnetspule lassen sich nach Belieben flexible Bewegungsmuster programmieren, um einen gezielten lateralen und vertikalen Transport der in dem Hybridisierungs- Flüssigkeitsfilm gelösten Target-Moleküle zu ermöglichen.Thus, the single magnetic coil above the chip serves to cause the magnetic particles to move upwards. If this coil is no longer magnetic by switching off the current, the magnetic particles begin to sink again. A drop in the same path as the lift path is prevented by the fact that the micro-magnetic coils of the micro-magnetic matrix below the chip, eg wave-like magnetized and thus the magnetic particles are displaced sideways during their sinking and thus ultimately a kind of oblique flow in the reaction liquid is induced, by which the target molecules more moved and thus more probe molecules are supplied. Thus, by means of the structure provided according to the invention, the particles are each moved laterally in the direction of the center of the correspondingly connected micro-magnetic matrix. By varying the duration and relative strength of the pulses from each adjacent micro-magnetic coil and the magnetic coil located above the chip, flexible motion patterns can be programmed as desired to enable targeted lateral and vertical transport of the target molecules dissolved in the hybridization liquid film ,
Die erfindungsgemäß vorgesehene Anordnung gestattet im Gegensatz zu den eingangs erwähnten Anordnungen gemäß dem Stand der Technik, welche sich mit der ungerichteten oder höchstens in einer Richtung erfolgenden Durchmischung von Flüssigkeitsfilmen befassen, eine vorprogrammierte, ganz gezielte Bewegung der eingesetzten Mikro-Magnetpartikel. Im Gegensatz zu den erwähnten Patenten Veröffentlichungen kann mit der erfindungsgemäßen Einrichtung insbesondere das Verklumpen und das Ansammeln der Magnetpartikel verhindert werden. Außerdem bietet die Erfindung den Vorteil, dass sie problemlos mit schon etablierten, bestehenden Bioanalyse-Arrays kombinierbar ist und weiters, dass bei Integration der Magnetspulen in einen entsprechenden Träger die Temperatur an der Grenzfläche zum Bio-Chip präzise eingestellt werden kann, was durch einen später noch zu erörternden integrierten Kühl/Heizkreislauf ermöglicht wird. Die Vorrichtung gemäß der Erfindung unterscheidet sich dadurch insbesondere von einer Anordnung, welche ausschließlich auf mögliche Prozessschritte in einem integrierten Mikrofluidik-Biochip abzielt und für eine Nachrüstung schon existierender DNA-Mikro-Arrays nicht geeignet ist.The inventively provided arrangement allows in contrast to the aforementioned arrangements according to the prior art, which deal with the non-directional or at most in one direction mixing of liquid films, a preprogrammed, very targeted movement of the micro-magnetic particles used. In contrast to the cited patents publications can be prevented with the device according to the invention, in particular the clumping and accumulation of the magnetic particles. In addition, the invention has the advantage that it can be easily combined with already established, existing bio-analysis arrays and further, that when integrating the magnetic coils in a corresponding carrier, the temperature at the interface with the bio-chip can be precisely adjusted, which later to be discussed integrated cooling / heating circuit is made possible. The device according to the invention thus differs in particular from an arrangement which exclusively aims at possible process steps in an integrated microfluidic biochip and is not suitable for retrofitting already existing DNA microarrays.
Zum Nachweis der Wirkungsweise der erfindungsgemäßen Vorrichtung wurden Hybridisierungs-Experimente auf gleichartigen DNA-Bio-Chips mit Unterstützung durch die erfindungsgemäße Vorrichtung und parallel dazu auf konventionelle Art durchgeführt. Hierbei waren konstante Temperaturverhältnisse (65 °C), Hybridisierungszeiten (25 min.) und Auswerteverfahren in den jeweiligen Experimenten sichergestellt; es zeigten sich beim Einsatz der erfindungsgemäßen Vorrichtung über sämtliche Experimente ein mittlerer Signalgewinn von etwa 150 % Signalgewinn =((Vorrichtungssignale- Referenzsignale)/Referenzsignale).100) gegenüber dem entsprechenden Ergebnissen bei herkömmlicher Hybridisierung.To demonstrate the mode of action of the device according to the invention, hybridization experiments were carried out on similar DNA biochips with support by the device according to the invention and in a conventional manner in parallel thereto. Constant temperature conditions (65 ° C), hybridization times (25 min.) And evaluation procedures in the respective experiments were ensured; When using the device according to the invention over all experiments, a mean signal gain of about 150% signal gain = ((device signal reference signals) / reference signals) .100) was found over the corresponding results in conventional hybridization.
Gemäß einer vorteilhaften Ausführungsform der Erfindung ist eine Ansteuerungs- Einrichtung für die Mikro-Magnetspulen einerseits und die Einzel-Magnetspule andererseits gemäß A n s p r u c h 2 vorgesehen, mittels welcher jede einzelne Mikro- Magnetspule individuell ansteuerbar ist und mittels welcher jedes zeitabhängige Magnetisierungs-Muster auf die Magnet-Matrixfläche aufprägbar ist. Insbesondere im Sinne einer ungestörten, optischen Kontrolle ist eineAccording to an advantageous embodiment of the invention, a driving device for the micro-magnetic coils on the one hand and the single solenoid on the other hand according to A nspruch 2 is provided by means of which each individual micro-solenoid is individually controlled and by means of which each time-dependent magnetization pattern on the magnet Matrix surface can be imprinted. Especially in terms of undisturbed, optical control is a
Ausführungsform der Einzelmagnetspule der erfindungsgemäßen Vorrichtung ohne Kern, also mit freigelassener mittlerer Ausnehmung, gemäß A n s p r u c h 3 günstig, wodurch eine freie Sicht auf das Reaktionsgeschehen, insbesondere auf den Bioanalyse-Chip bzw. -Array ermöglicht ist.Embodiment of the single magnetic coil of the device according to the invention without core, so with released middle recess, according to A n s p r u c h 3 low, whereby a clear view of the reaction process, in particular on the bio-analysis chip or -Array is enabled.
Zur Erhöhung der Effektivität der Anordnung der Mikro-Magnetspulen in der Magnetspulen-Matrix dienen die Merkmale des A n s p r u c h s 4 .To enhance the effectiveness of the arrangement of the micro-magnetic coils in the magnetic coil matrix serve the characteristics of the A n s p r u c h s 4th
Weiters ist eine Anordnung der Analyse-Vorrichtung in einer Art klimatisierter Kammer gemäß A n s p r u c h 5, insbesondere im Hinblick auf genau einzuhaltende stabile Umgebungsbedingungen bevorzugt.Furthermore, an arrangement of the analysis device in a kind of air-conditioned chamber according to A n s p r u c h 5, in particular with regard to stable environmental conditions to be observed exactly preferred.
Der A n s p r u c h 6 betrifft eine konkrete Anordnung der Komponenten der erfindungsgemäßen Vorrichtung.The invention relates to a specific arrangement of the components of the device according to the invention.
Schließlich hat der A n s p r u c h 7 eine bevorzugte Ausführungsvariante der Vorrichtung mit einer Aufnahmekammer für das Reaktionsgefäß, insbesondere für den Bio-Analysearray-Mikrochip, zum Gegenstand.Finally, the invention relates to a preferred embodiment of the device with a receiving chamber for the reaction vessel, in particular for the bio-analysis array microchip.
Anhand der Zeichnung wird die Erfindung näher erläutert:Reference to the drawing, the invention is explained in more detail:
Es zeigen die Fig. 1 die grundsätzliche Anordnung der wesentlichen Komponenten der erfindungsgemäßen Vorrichtung, die Fig. 2 schematisch die Vorgänge während der1 shows the basic arrangement of the essential components of the device according to the invention, the Fig. 2 shows schematically the processes during
Magnetisierung auf dem Objektträger, die Fig. 3 schematisch die Steuerung der neuen Vorrichtung, die Fig. 4 ein Beispiel für eine reale Ausführungsform der neuen Vorrichtung, die Fig. 5 schematisch ein Diagramm des Weges, welchen ein Magnetpartikel innerhalb der Probeflüssigkeit zurücklegt, die Fig. 6a ein Diagramm, welches die Signalsteigerung bei Einsatz der neuen Vorrichtung im Vergleich zeigt und die Fig. 6b ein Diagramm, in welchem die Signalsteigerung in Abhängigkeit von der Hybridisierungszeit ebenfalls im Vergleich zum Stand der Technik dargestellt ist:Magnetization on the slide, Fig. 3 shows schematically the control of the new device, Fig. 4 shows an example of a real embodiment of the new device, Fig. 5 shows schematically a diagram of the path which a magnetic particle travels within the sample liquid, Fig 6a shows a diagram which shows the signal increase when using the new device in comparison, and Fig. 6b shows a diagram in which the signal increase as a function of the hybridization time is also shown in comparison with the prior art:
In der Schrägrissdarstellung der Fig. 1 ist der prinzipielle Aufbau der erfindungsgemäßen Vorrichtung 1 für eine magnet-induzierte Intensivierung der Kontakte zwischen zwei Reaktions-, insbesondere Bindungspartnern, wie z.B. Target-Molekül und Sonden-Molekül gezeigt. Auf dem Objektträger 4 aus Glas ist unterhalb des Deckgläschens 5 derIn the oblique view of Fig. 1 is the basic structure of the device 1 according to the invention for a magnet-induced intensification of the contacts between two reaction, in particular binding partners, such. Target molecule and probe molecule shown. On the glass slide 4 is below the coverslip 5 of the
Bioanalyse-Array 6 mit in regelmäßig aufgetragenen feinen Spots mit den Sonden- Molekülen bzw. der Bio-Chip 6 angeordnet. Zwischen Objektträger 4 und Deckgläschen 5 befindet sich die Reaktions-, insbesondere Hybridisierungsflüssigkeit mit den Target- Molekülen und den für die Agitation der Flüssigkeit vorgesehenen Mikro-Magnetpartikel. In der Fig. 1 oberhalb dieser Anordnung ist der hier flachringartige bzw. toriodförmige Einzelmagnetspule 3 ohne Kern, also mit freier zentraler Öffnung 36 angeordnet, durch welche die Durchsicht auf die Reaktion auf dem Objektträger 4 bzw. unterhalb des Deckgläschens 5 freigehalten ist.Bioanalysis array 6 with arranged in regularly applied fine spots with the probe molecules or the bio-chip 6. Between slide 4 and coverslip 5 is the reaction, in particular hybridization liquid with the target molecules and provided for the agitation of the liquid micro-magnetic particles. In Fig. 1 above this arrangement, the here flat ring-like or toriodförmige single magnetic coil 3 without core, so with a free central opening 36th arranged, through which the review of the reaction on the slide 4 and below the coverslip 5 is kept free.
Unterhalb des Bio-Chips 6 und seinem Nahbereich ist unterhalb des Objektträgers 4 eine große Anzahl von individuell mit Magnetisierungsstrom versorgbaren Mikro- Magnetspulen 2 mit Spulenkernen 21 , vorzugsweise in einer Sechseck-Matrix 20 angeordnet.Below the bio-chip 6 and its vicinity, a large number of micro-magnet coils 2 with coil cores 21, preferably in a hexagonal matrix 20, can be arranged underneath the microscope slide 4.
Die schematische Darstellung der Fig. 2 zeigt - bei sonst gleichbleibenden Bezugszeichenbedeutungen - wie sich zwischen dem Objektträger 4 mit den Spots mit den Sonden-Molekülen 63 des Bio-Chips 6 die Reaktionsflüssigkeit 70 befindet, und wie sich in derselben - von den vorher erläuterten Elektro-Magneten 2 bzw. 20, 3 und deren variablen Magnetfeldern beaufschlagten, die hier kreisrund dargestellten Magnetpartikel 75 etwa in den durch Pfeile symbolisierten Bewegungsrichtungen bewegen und die durch Sternchen symbolisierten - Fluoreszenzfarbstoff 72 beaufschlagten Target-Moleküle 73 mit den immobilisierten Sonden-Molekülen 63 in Kontakt bringen und dort wesentlich öfter binden, als dies z.B. durch Diffusion erreichbar wäre.The schematic representation of Fig. 2 shows - with otherwise constant reference numerals - as between the slide 4 with the spots with the probe molecules 63 of the bio-chip 6, the reaction liquid 70 is located, and as in the same - from the previously described electro Magnets 2 and 20, 3 and their variable magnetic fields acted upon, the magnetic particles 75 shown here circular move approximately in the symbolized by arrows movement directions and the symbolized by asterisks - fluorescent dye 72 acted upon target molecules 73 with the immobilized probe molecules 63 in contact bring and bind there much more often than this, for example would be achievable by diffusion.
Die in der Fig. 3 - bei sonst gleichbleibenden Bezugszeichenbedeutungen - sehr schematisch dargestellten Steuerungs-Einrichtung 8 dient zur individuellen Anspeisung der Mikro-Magnetspulen 2 der Mikro-Magnetmatrix 20 mit variablen Magnetisierungsströmen sowie zur ebenfalls variablen Anspeisung der Einzel- Magnetspule 3 mit Strom.The control device 8 shown very schematically in FIG. 3 - with otherwise identical reference numerals - is used for individual energization of the micro-magnetic coils 2 of the micro-magnetic matrix 20 with variable magnetizing currents as well as for the likewise variable supply of the individual magnetic coil 3 with current.
Die Zentral-Steuerungseinheit (PC-Control) 81 ist mit einer Steuereinheit, z.B. D/A- card 82 verbunden, welche einerseits mit der Stromversorgungseinheit (Power-Supply 1) 83 für die Mikro-Magnetspulen-Matrix 20 und anderseits mit der Stromversorgungseinheit (Power Supply 2) 84 für die Einzel-Magnetspule 3 verbunden ist. Die Stromversorgungseinheit (Power Supply 1) 83 ist mit einer - selbst direkt mit der D/A-Card 82 verbundenen Relaismatrix-Einheit 85 verbunden, von den aus die zeitabhängig variable individuelle Stromversorgung jeder einzelnen mit derselben verbundenen einzelnen Mikromagnetspule 2 der Magnetmatrix 20 gemäß einem von der Zentral-Steuereinheit 81 gelieferten Programm erfolgt. Vorgesehen ist weiters auch eine Kontrolle der Temperatur an der Probe. Diese erfolgt mittels einer direkt mit der Zentral-Steuereinheit 81 verbundenen Temperatur- Kontrolleinheit (Thermocontrol) 86, welche von hier nicht gezeigten Thermosensoren, welche im Probebereich bzw. in Nähe der Magnete 2, 20 und 3 angeordnet sind, mit den aktuellen Temperaturdaten versorgt werden. Die Fig. 4 zeigt - bei sonst gleichbleibenden Bezugszeichenbedeutungen - jeweils in Schnittansichten von der Seite und von oben, die reale Konfiguration der Komponenten in der neuen verbesserten Analyse-Vorrichtung 1. Es ist hier eine Deckeinheit Il und eine Grundblockeinheit I gezeigt, zwischen welchen der hier nicht gezeigte Objektträger mit der durchzuführenden Reaktion angeordnet wird.The central control unit (PC control) 81 is connected to a control unit, eg D / A card 82, which is connected on the one hand to the power supply unit (power supply 1) 83 for the micro-solenoid matrix 20 and on the other hand to the power supply unit (FIG. Power Supply 2) 84 for the single solenoid 3 is connected. The power supply unit (power supply 1) 83 is connected to a - connected directly to the D / A card 82 relay matrix unit 85, from which the time-dependent variable individual power supply of each individual connected to the same single micro-magnetic coil 2 of the magnetic matrix 20 according to a is performed by the central control unit 81 program. It is also planned to control the temperature of the sample. This takes place by means of a temperature control unit (thermocontrol) 86 which is connected directly to the central control unit 81 and which is supplied with the current temperature data by thermosensors, not shown here, which are arranged in the sample area or in the vicinity of the magnets 2, 20 and 3 , Fig. 4 shows - with otherwise identical reference numerals - in each case in sectional views from the side and from above, the real configuration of the components in the new improved analysis device 1. Here is a cover unit II and a Basic block unit I shown, between which the slide not shown here is arranged with the reaction to be carried out.
Die Grundblockeinheit I besteht - wie aus Fig. 4a ersichtlich - aus einem von Kühl/The basic block unit I consists - as can be seen from FIG.
Heizmediumskanälen 22, 22' durchzogenen Aiuminiumblock 21 , in welchem mittig die Magnetspulen-Matrix 20 mit den Mikro-Magnetspulen 2 angeordnet ist. Unterhalb dieserHeizmediumskanälen 22, 22 'traversed Aiuminiumblock 21, in which the center of the magnetic coil array 20 is arranged with the micro-magnetic coils 2. Below this
Matrix 20 ist ein weiterer kleiner, ebenfalls von einem Kühl- bzw. Heizmediumskanal 225 durchzogener Aluminiumblock 25 angeordnet.Matrix 20 is another small, also from a cooling or Heizmediumskanal 225 traversed aluminum block 25 is arranged.
Den Verlauf der Kühl/Heizmediumskanäle 22, 22' im Grundriss erläutert die Fig. 4b, in welcher die Zu- und Abflüsse durch Pfeile gekennzeichnet sind. Durch die Fertigung der Kanäle im Block 21 bedingte Öffnungen sind mit Stopfen verschlossen.The course of the cooling / Heizmediumskanäle 22, 22 'in plan explains the Fig. 4b, in which the inflows and outflows are indicated by arrows. Due to the production of the channels in block 21 conditional openings are closed with plugs.
Nach oben hin abgedeckt ist der Aluminiumblock 21 mit einer dünnen Folie 23, auf welcher randseitig umlaufende Gummiringe 26,27 aufliegen, innerhalb welcher sich ein Raum 230 für die Platzierung der Probe befindet.Covered at the top is the aluminum block 21 with a thin film 23, on which peripherally surrounding rubber rings 26,27 rest, within which there is a space 230 for the placement of the sample.
Die in der Fig. 4c gezeigte Deckeinheit Il umfasst einen - hier mit der mittig eine Ausnehmung 34 aufweisenden Edelstahldeckfläche 35 - nach oben hin begrenzten Aluminiumblock 31 , welcher ebenfalls von Kühl/Heizmediumskanälen 32 durchzogen ist und in welchem die ringförmige Einzel-Magnetspule 3 untergebracht ist.The cover unit II shown in Fig. 4c comprises a - here with the center a recess 34 having stainless steel top surface 35 - limited upwards aluminum block 31, which is also crossed by cooling / Heizmediumskanälen 32 and in which the annular single solenoid 3 is housed ,
Der Aluminiumblock 31 ist von einer mittigen Öffnung 34' durchsetzt, durch welche der Blick auf die Probe freigehalten ist, und nach unten hin mit einer Glasplatte 33 abgeschlossen.The aluminum block 31 is penetrated by a central opening 34 ', through which the view of the sample is kept free, and closed at the bottom with a glass plate 33.
Die Temperaturkontrolle erfolgt mittels Thermoelementen 300, welche die aktuellen Temperaturdaten an die schon oben genannte Thermokontrolleinheit abgeben.The temperature control is carried out by means of thermocouples 300, which deliver the current temperature data to the above-mentioned thermal control unit.
Die Fig. 4d ermöglicht eine Übersicht über den Verlauf derFIG. 4d provides an overview of the course of the
Kühl/Heizmediumskanäle 32 im Aluminiumblock 31 der Deckeinheit Il in Draufsicht. Die Fig. 5 zeigt - bei sonst gleichbleibenden Bezugszeichenbedeutungen - schematisch den Weg- bzw. Bewegungsverlauf der Mirko-Magnetpartikel 75 imCooling / heating medium channels 32 in the aluminum block 31 of the cover unit II in plan view. Fig. 5 shows - with otherwise constant reference numerals - schematically the path or course of motion of the micro-magnetic particles 75 in
Flüssigkeitsfilm 70 zwischen Objektträger 4 bzw. Bio-Chip 6 und Deckglas 5. BeiLiquid film 70 between slide 4 or Bio-chip 6 and cover glass 5. Bei
Aktivieren der oberen Einzel-Magnetspule 3 wird das einzelne Magnetpartikel 75 entlangActivation of the upper single solenoid 3, the single magnetic particle 75 along
Bahn A etwa senkrecht aufwärts gezogen und durch das Magnetfeld einer etwas außerhalb der Aufstiegs-Bahn A liegenden jetzt magnetisierten Mikro-Magnetspule 2 und beim - durch Abschalten der Einzel-Magnetspule 3 erfolgenden Absinken wird dasPath A pulled approximately vertically upwards and by the magnetic field of a slightly out of the ascending path A lying now magnetized micro-magnetic coil 2 and - taking place by switching off the single solenoid 3 dropping is the
Magnetpartikel 75 seitlich abgelenkt und folgt dann etwa der Bahn B usw.Magnetic particle 75 deflected laterally and then follows about the web B, etc.
In der Fig. 6a und 6b sind die mit der erfindungsgemäßen Vorrichtung erreichten Signalsteigerungen als Funktion der Konzentration an magnetischen Partikeln nach der Hybridisierung gezeigt: Auf der Ordinate des Diagramms Fig.δaist die Intensität der Floureszenz-Indikation und auf der Abzisse ist die Konzentration der Magnetparikel M-PVA 13 bead (5-8 μm) (Chemagen AG, Arnold-Sommerfeld-Ring 2, D-52499 Baesweiler) in μg/μl aufgetragen.FIGS. 6a and 6b show the signal increases achieved with the device according to the invention as a function of the concentration of magnetic particles after hybridization: On the ordinate of the diagram Fig. 6a is the intensity of the fluorescence indication and on the abscissa is the concentration of the magnetic particles M-PVA 13 bead (5-8 μm) (Chemagen AG, Arnold-Sommerfeld-Ring 2, D-52499 Baesweiler) in μg / μl.
Die in Quadratform dargestellten Signalwerte wurden mit der beschriebenen Vorrichtung erzielt, die mit Kreuzen dargestellten auf konventionelle Weise. Es zeigt sich ein hoher mittlerer Signalgewinn. Die Fig. 6a zeigt weiters in der linken Spalte unten dieThe square-wave signal values were obtained with the described device, cross-hatched, in a conventional manner. It shows a high mean signal gain. Fig. 6a further shows in the left column below the
Referenzsignale der DNA-Sonde Ec. faecium 2 als kleine Kreuze. Der entsprechendeReference signals of the DNA probe Ec. faecium 2 as small crosses. The corresponding
Mittelwert ist in der rechten Spalte als kleines rotes Kreuz eingezeichnet. In der mittlerenMean is shown in the right column as a small red cross. In the middle
Spalte sind zu vier unterschiedlichen bead-Konzentrationen die Signale der Sonde Ec. faecium 2 bei Einsatz der neuen Vorrichtung dargestellt. Jeder der hier gezeigtenColumns are at four different bead concentrations the signals of the probe Ec. Faecium 2 shown when using the new device. Everyone shown here
Datenpunkte (kleine Quadrate) entspricht einem Experiment, die Fehlerbalken ergeben sich dabei durch die Auswertung von je 6 Replikaten der Sonde Ec. faecium 2 proData points (small squares) correspond to an experiment, the error bars result from the evaluation of 6 replicates of the probe Ec. faecium 2 per
Experiment. Die Hybridisierungszeit betrug in den jeweiligen Experimenten 25 min.Experiment. The hybridization time in the respective experiments was 25 min.
Es zeigte sich bei Einsatz der neuen Vorrichtung bei dieser Sonde über sämtliche Experimente ein mittlerer Signalgewinn von ca. 150 %, gegenüber einer herkömmlich durchgeführten Hybridisierung. Die Berechnung erfolgte durch arithmetisches Mitteln der Signale für die DNA-Sonde Ec. faecium 2 bei unterschiedlichen bead-Konzentrationen.It was found when using the new device in this probe over all experiments, a mean signal gain of about 150%, compared to a conventionally performed hybridization. The calculation was carried out by arithmetic means of the signals for the DNA probe Ec. Faecium 2 at different bead concentrations.
In der Fig. 6b sind - bei sonst gleichbleibenden Bezugszeichenbedeutungen - ebenfalls die unter Einsatz der erfindungsgemäßen Vorrichtung erzielten Intensitäten I der Fluoreszenz-Signale den Intensitäten der auf bisher bekannter Weise erhaltenen Signalen als Funktion der Hybridisierungszeit gegenübergestellt.In FIG. 6b, the intensities I of the fluorescence signals obtained using the device according to the invention are also compared with the intensities of the signals obtained in a previously known manner as a function of the hybridization time, with otherwise identical reference numerals.
Hier ist auf der Abzisse die Hybridisierungszeit in min aufgetragen und die Konzentration an Mikro-Magnetpartikeln bzw. -beads M-PVA 13 bead 5-8 μm ist mit [1,8 μg/μl ] konstant gehalten. Es zeigt sich schon nach 5 min ein sehr großer Signalgewinn bei Einsatz der erfindungsgemäßen Vorrichtung.Here, the hybridization time in minutes is plotted on the abscissa and the concentration of micro-magnetic particles or beads M-PVA 13 bead 5-8 μm is kept constant at [1.8 μg / μl]. It shows up after 5 min, a very large signal gain when using the device according to the invention.
(1) Advalytix AG, Eugen-Sänger-Straße 53.0, D-85649 Brunnthal, www.advalytix.de ,(1) Advalytix AG, Eugen-Sänger-Strasse 53.0, D-85649 Brunnthal, www.advalytix.de,
SlideBooster SB400 (2) R.H. Liu et.al. Bubble-induced acoustic micromixing, Lab Chip, 2002, 2, 151-157SlideBooster SB400 (2) R.H. Liu et al. Bubble-induced acoustic micromixing, Lab Chip, 2002, 2, 151-157
(3) WO 94/28396(3) WO 94/28396
(4) GB 876,070 A(4) GB 876,070 A
(5) WO 00/09991 A1(5) WO 00/09991 A1
(6) EP 0 240 862 A1 (7) WO 97/02357 A1(6) EP 0 240 862 A1 (7) WO 97/02357 A1
(8) US 6,806,050 B2 (8) US 6,806,050 B2

Claims

Patentansprüche: claims:
1. Vorrichtung zur Steigerung der Kontaktfrequenz bzw. -häufigkeit zwischen zwei miteinander zur Bindung befähigten Reaktionspartnern, vorzugsweise zwischen Analysator-Molekül bzw. -Molekülteil und Analyt-Molekül bzw. -Molekülteil, insbesondere zur Steigerung der Bindungseffektivität und zur Reduktion der für einen Nachweis erforderlichen Bindungszeiten in Bioanalyse-Arrays, wie z.B. DNA/RNA-, Protein- oder Immuno-Mikroarrays mit Hilfe von - mittels beidseitig des Reaktionsvolumens bzw. Reaktionsfluidfilms angeordneter, mit veränderlichen Strömen anspeisbarer Elektromagneten (3, 2, 20) - extern generierten Magnetfeldern berührungslos im fluiden Reaktionsmedium, insbesondere mit in der den Bioanalyse-Array umgebenden Flüssigkeit gezielt in Bewegung gesetzten para-, superpara- oder ferromagnetischen, sphärisch oder unregelmäßig geformten, gegebenenfalls mit einem der Reaktionspartner gecoateten, Mikro- oder Nanopartikeln (75) d a d u r c h g e k e n n z e i c h n e t , dass auf einer Seite des Reaktionsgefäßes bzw. Reaktionsfluidfilms, insbesondere eines Objektträgers (4) mit - den bzw. die Reaktionspartner (73) und die Reaktionsflüssigkeit (70) und vorzugsweise ein Mikro-Bioanalyse-Array (6) abdeckendem - transparentem Deckglas (5), in der unmittelbarer Nähe eine zweidimensional flächige Matrix (20) bzw. ein Array mit einer Mehrzahl von individuell mit Magnetisierstrom jeweils vorgegebener, zeitabhängig variabler Stärke und/oder Spannung - einem jeweils gewünschten zeitabhängig variablen bzw. veränderlichen Magnetisierungs- bzw. Feldstärke-Muster entsprechend - anspeisbaren Miniatur- bzw. Milli-Magnetspulen (2) gegebenenfalls mit oder ohne feldverstärkendem Kern (21), angeordnet ist und dass auf der anderen Seite des Reaktionsgefäßes, insbesondere Objektträger (4) mit dem Mikro-Bio-Analysearray (6), in dessen unmittelbarer Nähe nur eine, ebenfalls mit variablen Magnetisierstrom anspeisbare Magnetspule (3) positioniert ist, deren Magnetfeld das gesamte Reaktionsgefäß oder wesentliche Teile desselben, insbesondere den gesamten (Mikro)-Bioanalysearray (6), durchsetzt.1. A device for increasing the frequency or frequency of contact between two reactants capable of binding with one another, preferably between analyzer molecule or molecule part and analyte molecule or molecule part, in particular for increasing the binding efficiency and for reducing the amount required for detection Binding times in bioanalysis arrays, such as DNA / RNA, protein or immuno-microarrays by means of - arranged on both sides of the reaction volume or reaction fluid film, with variable currents anspeisbarer electromagnets (3, 2, 20) - externally generated magnetic fields without contact in the fluid reaction medium, in particular with in the Bio-analytic array surrounding liquid specifically set in motion para-, superpara- or ferromagnetic, spherical or irregularly shaped, optionally coated with one of the reactants, micro or nanoparticles (75) characterized in that on one side of the reaction vessel or reaction fluid film, in particular a Object carrier (4) with - the or the reaction partners (73) and the reaction liquid (70) and preferably a micro-bioanalysis array (6) covering - transparent cover glass (5), in the immediate vicinity of a two-dimensional area matrix (20) or an array with a plurality of individually with Magnetizing each predetermined, time-dependent variable strength and / or voltage - a respectively desired time-dependent variable or variable magnetization or field strength pattern according to - anspeisbaren miniature or Milli magnetic coils (2) optionally with or without feldverstärkendem core (21), is arranged and that on the other side of the reaction vessel, in particular slide (4) with the micro-bio-analysis array (6), in the immediate vicinity only one, also with variable magnetizing anspeisbare magnetic coil (3) is positioned, the magnetic field of the entire Reaction vessel or essential parts thereof, in particular the entire (micro) -Bioanalysearray (6), interspersed.
2. Vorrichtung nach Anspruch 1 , d a d u r c h g e k e n n z e i c h n e t , dass sie eine Magneten-Ansteuerungseinrichtung (8) mit Zentral-Steuereinheit (81) aufweist, mittels welcher die Einzel-Magnetspule (3) und jede der Miniatur-Magnetspulen (2) der Magnetspulen-Matrix bzw. des Magnetspulen-Arrays (20) jeweils individuell und unabhängig von den anderen Miniatur-Magnetspulen, mit insbesondere hinsichtlich Stromart, wie DC oder AC, mit beliebiger Frequenz, Wellenform, Amplitude und/oder Phasenverschiebung, variablen Magnetisierungsstrom anspeisbar bzw. bestrombar sind, sodass - einem jeweils gewählten Programm entsprechend - jeweils örtlich, in allen drei Raumrichtungen, verschieden - eine ortsvariable Bewegung der Magnet-Partikel (75) mit jeweils gewünschter Bewegungsrichtung, -bahn und -geschwindigkeit in der Reaktionsflüssigkeit im Reaktionsfluidfilm bzw. in der den (Mikro-)Bioanalyse-Array bzw. Bio-Chip (6) umgebenden Flüssigkeit induzierbar ist.2. Device according to claim 1, characterized in that it comprises a magnet drive means (8) with central control unit (81), by means of which the single solenoid coil (3) and each of the miniature magnetic coils (2) of the magnetic coil matrix or of the magnetic coil array (20) each individually and independently of the other miniature magnetic coils, in particular with respect to current, such as DC or AC, with any frequency, waveform, amplitude and / or Phase shift, variable magnetizing current are anspeisbar or energized, so - according to a selected program - each locally, in all three spatial directions, different - a spatially variable movement of the magnetic particles (75) each having the desired direction of movement, trajectory and speed in the Reaction fluid in the reaction fluid film or in the (micro) Bioanalyse array or bio-chip (6) surrounding liquid is inducible.
3. Vorrichtung nach einem der Ansprüche 1 oder 2, dadurch gekennzeichnet, dass die Einzel-Magnetspule (3) ringförmig und ohne Kern ausgebildet ist, um die Sicht auf das Reaktionsgefäß, insbesondere auf den (Mikro-)Bioanalyse-Array bzw. Mikro- Biochip (6), freizuhalten.3. Device according to one of claims 1 or 2, characterized in that the single magnetic coil (3) is annular and without a core formed to the view of the reaction vessel, in particular on the (micro) bioanalysis array or micro Biochip (6), keep clear.
4. Vorrichtung nach einem der Ansprüche 1 bis 3, dadurch gekennzeichnet, dass die Mikro-Magnetspulen (2) der Mikro-Magnetspulen-Matrix (20) mit möglichst kleinen Zwischenräumen zwischen einander mit jeweils kreisförmigem, quadratischem oder hexagonalem Querschnitt ausgebildet und in einer Quadrat-Matrix oder bienenwabenartigen Sechseck-Matrix (20) angeordnet sind.4. Device according to one of claims 1 to 3, characterized in that the micro-magnetic coils (2) of the micro-magnetic coil matrix (20) formed with the smallest possible spaces between each other with each circular, square or hexagonal cross-section and in a square Matrix or honeycomb hexagon matrix (20) are arranged.
5. Vorrichtung nach einem der Ansprüche 1 bis 4, dadurch gekennzeichnet, dass sie insgesamt in einer Kammer angeordnet ist, in welcher die Feuchte des das Reagenzgefäß, insbesondere den (Mikro-)Bioanalyse-Array (6), umgebenden Gases, sowie vorzugsweise die bzw. dessen Temperatur und gegebenenfalls Druck kontrollier- und einstellbar ist.5. Device according to one of claims 1 to 4, characterized in that it is arranged in a total chamber in which the humidity of the reagent vessel, in particular the (micro) bioanalysis array (6), surrounding gas, and preferably the or whose temperature and optionally pressure can be controlled and adjusted.
6. Vorrichtung nach einem der Ansprüche 1 bis 5 dadurch gekennzeichnet,6. Device according to one of claims 1 to 5 characterized in that
- dass die Einzel-Magnetspule (3) in einem Block (31) aus gut wärmeleitfähigem Material, insbesondere aus Aluminium, angeordnet ist und dass in demselben ein diese Einzel- Magnetspule (3) im Nahbereich ihrer dem Reaktionsgefäß, insbesondere dem (Mikro-)Bioanalyse-Array (6), nahen Oberfläche außen umringender und ein vorzugsweise gesonderter, oberhalb derselben im Nahbereich von deren Innenrand angeordneten, mit einer Kühl- oder Heizflüssigkeit durchströmbarer Fluidmediums-Kanal (32, 32') vorgesehen ist. - dass gegebenenfalls auch die Mikro-Magnetspulen-Matrix (20) in einem Aluminiumblock oder -tisch (21) angeordnet ist und seitlich rundum von einem mit einer Kühl- oder Heizflüssigkeit durchströmbaren Fluidmediums-Kanal (22, 22') umgeben ist und- That the single magnetic coil (3) is arranged in a block (31) made of good thermally conductive material, in particular aluminum, and that in the same a single solenoid coil (3) in the vicinity of their the reaction vessel, in particular the (micro-) Bioanalysis array (6), the surrounding surface on the outside surrounding and preferably a separate, arranged above the same in the vicinity of the inner edge, with a cooling or heating fluid permeable fluid medium channel (32, 32 ') is provided. - That, where appropriate, the micro-magnetic coil matrix (20) in an aluminum block or table (21) is arranged and laterally surrounded all around by a through-flow of a cooling or heating fluid fluid channel (22, 22 ') and
- dass gegebenenfalls zusätzlich unterhalb der Mikro-Magnetspulen-Matrix (20) ein Kühlkanal-System (225) in einem gesonderten Metallblock (25) angeordnet ist.- That optionally additionally below the micro-magnetic coil matrix (20), a cooling channel system (225) in a separate metal block (25) is arranged.
7. Vorrichtung nach Anspruch 6, d a d u r c h g e k e n n z e i c h n e t ,7. The device according to claim 6, wherein said device is a device of claim 6, wherein
- dass der Metall-, insbesondere Aluminiumblock (31) mit der Einzelmagnetspule (3) zum Reaktionsgefäß, insbesondere zum Bioanalyse-Array (6) bzw. Mikro-Biochip, hin mit einer- That the metal, in particular aluminum block (31) with the single magnetic coil (3) to the reaction vessel, in particular to the bio-analysis array (6) or micro-biochip, out with a
Glasplatte (33) abgeschlossen ist,Glass plate (33) is completed,
- dass gegebenenfalls der Metall-, insbesondere Aluminiumblock (21) mit der Mikro- Magnetspulen-Matrix (20) mittels Aluminium- oder Kunststoff-Deckfolie (23) abgeschlossen ist und - dass gegebenenfalls die beiden Metall-, insbesondere Aluminiumblöcke (21 , 31) mittels mindestens eines randnah angeordneten geschlossenen Gummi-Ringelements (26, 27) zwischen der oben genannten Glasplatte (23) und der Deckfolie (23) unter Ausbildung einer von der Umgebung und deren Einflüssen abgeschotteten Probenkammer (230) voneinander beabstandet gehalten sind. - That optionally the metal, in particular aluminum block (21) with the micro-magnetic coil matrix (20) by means of aluminum or plastic cover film (23) is completed and - that optionally the two metal, in particular aluminum blocks (21, 31) by means of at least one rubber ring element (26, 27) arranged close to the edge between the above-mentioned glass plate (23) and the cover film (23), while keeping a sample chamber (230) sealed off from the environment and its influences.
EP07718377A 2006-04-13 2007-04-11 Apparatus for enhancing the reaction efficiency, especially the binding efficiency, between molecules and molecular moieties Not-in-force EP2004317B1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
AT0064806A AT503573B1 (en) 2006-04-13 2006-04-13 DEVICE FOR INCREASING THE RESPONSE, ESPECIALLY THE CONNECTION EFFICIENCY BETWEEN MOLECULES AND MOLECULAR COMPONENTS
PCT/AT2007/000160 WO2007118261A1 (en) 2006-04-13 2007-04-11 Apparatus for enhancing the reaction efficiency, especially the binding efficiency, between molecules and molecular moieties

Publications (2)

Publication Number Publication Date
EP2004317A1 true EP2004317A1 (en) 2008-12-24
EP2004317B1 EP2004317B1 (en) 2010-08-25

Family

ID=38226496

Family Applications (1)

Application Number Title Priority Date Filing Date
EP07718377A Not-in-force EP2004317B1 (en) 2006-04-13 2007-04-11 Apparatus for enhancing the reaction efficiency, especially the binding efficiency, between molecules and molecular moieties

Country Status (7)

Country Link
US (1) US20090325822A1 (en)
EP (1) EP2004317B1 (en)
CN (1) CN101541410A (en)
AT (2) AT503573B1 (en)
DE (1) DE502007004860D1 (en)
ES (1) ES2348943T3 (en)
WO (1) WO2007118261A1 (en)

Families Citing this family (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2009539100A (en) * 2006-06-02 2009-11-12 コーニンクレッカ フィリップス エレクトロニクス エヌ ヴィ Small electronic sensor device having cleaning means
CH700770A2 (en) 2009-04-15 2010-10-15 Philippe Saint Ger Ag A method for supporting and / or intensifying a physical and / or chemical reaction and a reaction device for performing the method.
US8264224B2 (en) * 2009-10-27 2012-09-11 University Of Seoul Industry Cooperation Foundation Detection of magnetic fields using nano-magnets
US8289022B2 (en) 2010-01-29 2012-10-16 University Of Seoul Industry Cooperation Foundation Magnetic resonance compatible magnetic field detection, based on diffuse reflectance of nano-magnet sets
CN102234611A (en) * 2010-04-27 2011-11-09 杭州绿洁水务科技有限公司 Control device applicable to comprehensive toxicity detection
DE102012210077A1 (en) 2012-06-15 2013-12-19 Siemens Aktiengesellschaft Method and device for labeling cells in a cell suspension
DE102013009773B4 (en) 2013-06-05 2016-02-11 Technische Universität Dresden Device and method for increasing the binding efficiency of binding capable target structures
CN103398252B (en) * 2013-08-07 2016-09-28 苏州扬清芯片科技有限公司 A kind of electromagnetic type chip connector
CN103439506B (en) * 2013-08-12 2015-04-15 杭州戈登生物科技有限公司 Rapid detection device and method for interaction of biomacromolecules
JP6908624B2 (en) * 2016-04-27 2021-07-28 ベンタナ メディカル システムズ， インコーポレイテッド Systems and methods for real-time volume control
CN106179544B (en) * 2016-07-14 2018-07-06 大连海事大学 Portable immunomagnetic beads three-dimensional hybrid device and application method based on micro-fluidic chip
CN107811747A (en) * 2017-10-31 2018-03-20 姚森 A kind of arthritis physiotherapy Diannuanbao of consistent heat generation

Family Cites Families (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5222808A (en) * 1992-04-10 1993-06-29 Biotrack, Inc. Capillary mixing device
CN1185492C (en) * 1999-03-15 2005-01-19 清华大学 Single-point strobed micro electromagnetic units array chip or electromagnetic biologic chip and application thereof
EP1263532A2 (en) * 2000-03-16 2002-12-11 Sri International Microlaboratory devices and methods
EP1287167A2 (en) * 2000-04-11 2003-03-05 Mats Bo Johan Nilsson Nucleic acid detection medium
DE60117556T2 (en) * 2000-06-21 2006-11-02 Bioarray Solutions Ltd. MULTI-ANALYTIC MOLECULAR ANALYSIS THROUGH THE USE OF APPLICATION SPECIFIC RAPID PARTICLE ARRAYS
WO2002012896A1 (en) * 2000-08-08 2002-02-14 Aviva Biosciences Corporation Methods for manipulating moieties in microfluidic systems
US20020166760A1 (en) * 2001-05-11 2002-11-14 Prentiss Mara G. Micromagentic systems and methods for microfluidics
US7195913B2 (en) * 2001-10-05 2007-03-27 Surmodics, Inc. Randomly ordered arrays and methods of making and using
JP2003248008A (en) * 2001-12-18 2003-09-05 Inst Of Physical & Chemical Res Method of stirring reaction liquid
WO2005072855A1 (en) * 2004-01-28 2005-08-11 Drexel University Magnetic fluid manipulators and methods for their use

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO2007118261A1 *

Also Published As

Publication number Publication date
ES2348943T3 (en) 2010-12-17
AT503573B1 (en) 2007-11-15
DE502007004860D1 (en) 2010-10-07
WO2007118261A1 (en) 2007-10-25
ATE478727T1 (en) 2010-09-15
AT503573A4 (en) 2007-11-15
EP2004317B1 (en) 2010-08-25
US20090325822A1 (en) 2009-12-31
CN101541410A (en) 2009-09-23

Similar Documents

Publication Publication Date Title
AT503573B1 (en) DEVICE FOR INCREASING THE RESPONSE, ESPECIALLY THE CONNECTION EFFICIENCY BETWEEN MOLECULES AND MOLECULAR COMPONENTS
US20190324026A1 (en) Droplet Actuator Devices and Methods Employing Magnetic Beads
EP1843854B1 (en) Device and method for the elimination of magnetic or magnetizable particles from a liquid
DE102009012169B3 (en) Apparatus and method for making a replica or derivative from an array of molecules and applications thereof
DE60110256T2 (en) METHOD FOR CONCENTRATING ANALYTES IN SAMPLES
DE60013848T2 (en) MICROFLUID DEVICES FOR THE CONTROLLED HANDLING OF SMALL VOLUMES
DE10156790A1 (en) Device and method for treating magnetic particles
CN103930210A (en) Microfluidic system
CA2424312A1 (en) Individually addressable micro-electromagnetic unit array chips in horizontal configurations
DE60023185T2 (en) APPARATUS AND METHOD FOR MIXING AND DISCONNECTING USING MAGNETIC PARTICLES
EP2030689A1 (en) Microplate support with magnets
EP1880766A1 (en) Analysis system based on porous material for highly-parallel single cell detection
DE60214155T2 (en) METHOD FOR ACCELERATING AND REINFORCING THE BINDING OF TARGET COMPONENTS TO RECEPTORS AND DEVICE THEREFOR
DE102010032203A1 (en) Method and apparatus for the passive separation and sorting of drops, in particular in a microfluidic system, by using non-optical markers for reactions within the drops
US10710079B2 (en) Electro-kinectic device for species exchange
WO2019057345A1 (en) Device and method for immobilising biomolecules using magnetic particles
DE10136008B4 (en) Method for analyzing macromolecules and method for producing an analysis device
EP2591086B1 (en) Method for active hybridization in microarrays with denaturing function
DE102020209001B4 (en) Lysis of a sample using magnetic elements and rotary relative movement
DE10231925A1 (en) Reactor system for use in (bio)chemical research, has a miniaturized reactor array, with a magnetic field array under and parallel to it, fitted with magnets to manipulate magnetic particles in reaction clusters at the reaction sites
DE10322942A1 (en) Device for positioning and discharging fluid compartments embedded in the separation medium
DE19955169A1 (en) Solenoids with air gap to influence magnetic particles in fluid in a titration plate, is useful for mixing, vibration, precipitation and filtration
EP1507591A1 (en) Method for transferring molecules from a chemically reacting first flow into an adjacent chemically second reacting flow
DE102004018971A1 (en) Microfluidic apparatus producing micro-flows through magnetohydrodynamic principles, subjects electrolyte fluid volume to magnetic and electrical fields
AU2013267077A1 (en) Droplet Actuator Devices And Methods Employing Magnetic Beads

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 20081029

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IS IT LI LT LU LV MC MT NL PL PT RO SE SI SK TR

RAP1 Party data changed (applicant data changed or rights of an application transferred)

Owner name: TECNET CAPITAL TECHNOLOGIEMANAGEMENT GMBH

Owner name: AUSTRIAN RESEARCH CENTERS GMBH-ARC

GRAP Despatch of communication of intention to grant a patent

Free format text: ORIGINAL CODE: EPIDOSNIGR1

GRAS Grant fee paid

Free format text: ORIGINAL CODE: EPIDOSNIGR3

GRAA (expected) grant

Free format text: ORIGINAL CODE: 0009210

AK Designated contracting states

Kind code of ref document: B1

Designated state(s): AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IS IT LI LT LU LV MC MT NL PL PT RO SE SI SK TR

REG Reference to a national code

Ref country code: GB

Ref legal event code: FG4D

Free format text: NOT ENGLISH

REG Reference to a national code

Ref country code: CH

Ref legal event code: EP

REG Reference to a national code

Ref country code: IE

Ref legal event code: FG4D

Free format text: LANGUAGE OF EP DOCUMENT: GERMAN

RAP2 Party data changed (patent owner data changed or rights of a patent transferred)

Owner name: TECNET CAPITAL TECHNOLOGIEMANAGEMENT GMBH

Owner name: AIT AUSTRIAN INSTITUTE OF TECHNOLOGY GMBH

REF Corresponds to:

Ref document number: 502007004860

Country of ref document: DE

Date of ref document: 20101007

Kind code of ref document: P

REG Reference to a national code

Ref country code: CH

Ref legal event code: NV

Representative=s name: BUECHEL, VON REVY & PARTNER

REG Reference to a national code

Ref country code: NL

Ref legal event code: T3

REG Reference to a national code

Ref country code: ES

Ref legal event code: FG2A

Effective date: 20101203

LTIE Lt: invalidation of european patent or patent extension

Effective date: 20100825

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: LT

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20100825

Ref country code: FI

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20100825

REG Reference to a national code

Ref country code: NL

Ref legal event code: TD

Effective date: 20110209

Ref country code: NL

Ref legal event code: SD

Effective date: 20110209

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: BG

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20101125

Ref country code: PT

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20101227

Ref country code: PL

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20100825

Ref country code: IS

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20101225

Ref country code: CY

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20100825

Ref country code: SI

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20100825

REG Reference to a national code

Ref country code: GB

Ref legal event code: 732E

Free format text: REGISTERED BETWEEN 20110203 AND 20110209

REG Reference to a national code

Ref country code: IE

Ref legal event code: FD4D

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: LV

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20100825

Ref country code: SE

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20100825

Ref country code: GR

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20101126

REG Reference to a national code

Ref country code: DE

Ref legal event code: R081

Ref document number: 502007004860

Country of ref document: DE

Owner name: AIT AUSTRIAN INSTITUTE OF TECHNOLOGY GMBH, AT

Free format text: FORMER OWNER: AIT AUSTRIAN INSTITUTE OF TECHN, TECNET CAPITAL TECHNOLOGIEMANAG, , AT

Effective date: 20110215

Ref country code: DE

Ref legal event code: R081

Ref document number: 502007004860

Country of ref document: DE

Owner name: TECNET EQUITY NOE TECHNOLOGIEBETEILIGUNGS-INVE, AT

Free format text: FORMER OWNER: AIT AUSTRIAN INSTITUTE OF TECHN, TECNET CAPITAL TECHNOLOGIEMANAG, , AT

Effective date: 20110215

Ref country code: DE

Ref legal event code: R081

Ref document number: 502007004860

Country of ref document: DE

Owner name: AIT AUSTRIAN INSTITUTE OF TECHNOLOGY GMBH, AT

Free format text: FORMER OWNERS: AIT AUSTRIAN INSTITUTE OF TECHNOLOGY GMBH, WIEN, AT; TECNET CAPITAL TECHNOLOGIEMANAGEMENT GMBH, ST. POELTEN, AT

Effective date: 20110215

Ref country code: DE

Ref legal event code: R081

Ref document number: 502007004860

Country of ref document: DE

Owner name: TECNET EQUITY NOE TECHNOLOGIEBETEILIGUNGS-INVE, AT

Free format text: FORMER OWNERS: AIT AUSTRIAN INSTITUTE OF TECHNOLOGY GMBH, WIEN, AT; TECNET CAPITAL TECHNOLOGIEMANAGEMENT GMBH, ST. POELTEN, AT

Effective date: 20110215

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: DK

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20100825

Ref country code: IE

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20100825

REG Reference to a national code

Ref country code: FR

Ref legal event code: CD

Ref country code: FR

Ref legal event code: TQ

Ref country code: CH

Ref legal event code: PUEA

Owner name: AIT AUSTRIAN INSTITUTE OF TECHNOLOGY GMBH

Free format text: TECNET CAPITAL TECHNOLOGIEMANAGEMENT GMBH#WIRTSCHAFTSZENTRUM NIEDEROESTERREICH NIEDEROESTERREICHRING 2, HAUS B#3100 ST. POELTEN (AT) $ AIT AUSTRIAN INSTITUTE OF TECHNOLOGY GMBH#TECH GATE VIENNA WISSENSCH. U. TECH. PARK DONAU-CITY-STRASSE 1#1220 WIEN (AT) -TRANSFER TO- AIT AUSTRIAN INSTITUTE OF TECHNOLOGY GMBH#TECH GATE VIENNA WISSENSCH. U. TECH. PARK DONAU-CITY-STRASSE 1#1220 WIEN (AT) $ TECNET EQUITY NOE TECHNOLOGIEBETEILIGUNGS-INVEST GMBH#WIRTSCHAFTSZENTRUM NIEDEROESTERREICH NIEDEROESTERREICHRING 2, HAUS B, 5. OG#3100 ST. POELTEN (AT)

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: SK

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20100825

Ref country code: RO

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20100825

Ref country code: CZ

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20100825

Ref country code: EE

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20100825

PLBE No opposition filed within time limit

Free format text: ORIGINAL CODE: 0009261

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: NO OPPOSITION FILED WITHIN TIME LIMIT

BECH Be: change of holder

Owner name: AIT AUSTRIAN INSTITUTE OF TECHNOLOGY GMBH

Effective date: 20110706

Owner name: TECNET EQUITY NO TECHNOLOGIEBETEILIGUNGS-INVEST G.

Effective date: 20110706

26N No opposition filed

Effective date: 20110526

REG Reference to a national code

Ref country code: DE

Ref legal event code: R097

Ref document number: 502007004860

Country of ref document: DE

Effective date: 20110526

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: MC

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20110430

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: MT

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20100825

REG Reference to a national code

Ref country code: ES

Ref legal event code: PC2A

Owner name: TECNET EQUITY NOE TECHNOLOGIEBETEILIGUNGS-INVEST G

Effective date: 20120214

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: CH

Payment date: 20120420

Year of fee payment: 6

Ref country code: DE

Payment date: 20120420

Year of fee payment: 6

Ref country code: BE

Payment date: 20120418

Year of fee payment: 6

Ref country code: NL

Payment date: 20120425

Year of fee payment: 6

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: FR

Payment date: 20120507

Year of fee payment: 6

Ref country code: GB

Payment date: 20120419

Year of fee payment: 6

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: IT

Payment date: 20120420

Year of fee payment: 6

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: ES

Payment date: 20120425

Year of fee payment: 6

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: LU

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20110411

REG Reference to a national code

Ref country code: AT

Ref legal event code: MM01

Ref document number: 478727

Country of ref document: AT

Kind code of ref document: T

Effective date: 20120411

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: AT

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20120411

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: TR

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20100825

BERE Be: lapsed

Owner name: AIT AUSTRIAN INSTITUTE OF TECHNOLOGY GMBH

Effective date: 20130430

Owner name: TECNET EQUITY NO TECHNOLOGIEBETEILIGUNGS-INVEST G.

Effective date: 20130430

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: HU

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20100825

REG Reference to a national code

Ref country code: NL

Ref legal event code: V1

Effective date: 20131101

REG Reference to a national code

Ref country code: CH

Ref legal event code: PL

GBPC Gb: european patent ceased through non-payment of renewal fee

Effective date: 20130411

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: GB

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20130411

Ref country code: CH

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20130430

Ref country code: BE

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20130430

Ref country code: DE

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20131101

Ref country code: LI

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20130430

REG Reference to a national code

Ref country code: FR

Ref legal event code: ST

Effective date: 20131231

REG Reference to a national code

Ref country code: DE

Ref legal event code: R119

Ref document number: 502007004860

Country of ref document: DE

Effective date: 20131101

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: FR

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20130430

Ref country code: IT

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20130411

Ref country code: NL

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20131101

REG Reference to a national code

Ref country code: ES

Ref legal event code: FD2A

Effective date: 20140606

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: ES

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20130412