WO2013072110A1 - Élément filtrant microfluidique permettant de séparer des constituants d'un échantillon de fluide biologique - Google Patents

Élément filtrant microfluidique permettant de séparer des constituants d'un échantillon de fluide biologique Download PDF

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Publication number
WO2013072110A1
WO2013072110A1 PCT/EP2012/068901 EP2012068901W WO2013072110A1 WO 2013072110 A1 WO2013072110 A1 WO 2013072110A1 EP 2012068901 W EP2012068901 W EP 2012068901W WO 2013072110 A1 WO2013072110 A1 WO 2013072110A1
Authority
WO
WIPO (PCT)
Prior art keywords
filter element
microfluidic
filter
sample fluid
biological sample
Prior art date
Application number
PCT/EP2012/068901
Other languages
German (de)
English (en)
Inventor
Martina Daub
Thomas BRETTSCHNEIDER
Christian Dorrer
Original Assignee
Robert Bosch 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 Robert Bosch Gmbh filed Critical Robert Bosch Gmbh
Publication of WO2013072110A1 publication Critical patent/WO2013072110A1/fr

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Classifications

    • 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
    • B01L3/502753—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 characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation
    • 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
    • B01L3/502761—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 specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/06—Fluid handling related problems
    • B01L2200/0631—Purification arrangements, e.g. solid phase extraction [SPE]
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/06—Fluid handling related problems
    • B01L2200/0647—Handling flowable solids, e.g. microscopic beads, cells, particles
    • B01L2200/0668—Trapping microscopic beads
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/0627—Sensor or part of a sensor is integrated
    • B01L2300/0636—Integrated biosensor, microarrays
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/0681—Filter
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/40—Concentrating samples
    • G01N1/4077—Concentrating samples by other techniques involving separation of suspended solids
    • G01N2001/4088—Concentrating samples by other techniques involving separation of suspended solids filtration

Definitions

  • Microfluidic filter element for separating sample components from a biological sample fluid
  • the present invention relates to a microfluidic filter element for
  • compositions of the sample fluid can be separated from this. However, the often low concentration of these ingredients is one
  • the prior art uses a multiplicity of different filters which, on the one hand, are selected depending on the size of the constituents to be separated on the one hand and the remaining constituents contained in the sample fluid on the other hand.
  • the size of the components of the sample to be detected must therefore be sufficiently different from the remaining components of the sample liquid. In the case of bacteria in whole blood, this requirement is not met because the size of the bacteria is comparable to the size of the cellular components of the blood.
  • microchannel-based separation techniques which are the different hydrodynamic ones
  • GB-2401942 A discloses a filter element for use in a
  • Detection assay in which specific binding molecules are immobilized on the surface of a porous membrane.
  • the desired target components are trapped using the binding molecules and thus fixed on the underside of the membrane.
  • Another possibility is the separation of specific sample components from a sample fluid with the help of functionalized beads of a few
  • Micrometer diameter For this purpose, for example, antibodies are immobilized on the bead surface. After addition of the beads to the sample fluid, they bind to the associated antigens and in this way collect the desired sample constituents, such as, for example, pathogenic cells. The separation of the beads including the pathogenic cells from the sample fluid is then carried out by filtration or, in the case of magnetic beads, by external magnetic fields.
  • the subject of the present invention is thus a microfluidic
  • Filter element for separating certain sample components from a biological sample fluid, comprising
  • filter molecules immobilized on the filter material for the particular sample components, wherein the filter material is interspersed with the capture molecules.
  • the specific capture molecules such as antibodies or enzymes, but also polynucleic acids, polymers, etc. are on the surface of the
  • Filter material immobilized so that the desired sample components are bound to the capture molecules when flowing through the sample liquid.
  • the deposition of the sought sample components is distributed over the entire filter material, since the filter material is interspersed with the capture molecules and thus faces the sought sample components an extremely high number of possible binding sites.
  • the filter element of the invention operates extremely efficient and highly specific.
  • the pores of the filter material are dimensioned so that components of the sample liquid, which do not interact with the catcher molecules, can pass the filter substantially. A mechanical filtration through the pure filter material therefore plays a minor role.
  • sample constituents is understood to mean all desired specific constituents of the biological sample fluid which can be purposefully separated with the aid of the microfluidic filter element. These are in particular both pathogenic and non-pathogenic cells or viruses and their constituents, such as, for example, endotoxins, nucleic acids, proteins or other cellular constituents.
  • the sample components can be separated very efficiently and highly specifically from a biological sample fluid.
  • Sample components and the remaining components of the sample be comparable. This is partly due to the fact that a mechanical filtration is completely dispensed with and thus the pores of the filter can be chosen to be relatively large, so that a much higher throughput is possible. In addition, due to the large pores, filtration of large quantities of sample fluid is possible without the filter tending to clog.
  • Microfluidic filter element according to the invention thus represents a clear further development of known microfluidic filter systems, since they were hitherto only suitable for low throughput quantities.
  • the use of beads can be dispensed with.
  • the use of immobilized capture molecules that permeates the entire filter material enables highly specific filtration compared to conventional filters or methods based on hydrodynamic effects. On a dilution of the sample fluid in advance of the filtration can be dispensed with.
  • the binding of the particular sample components to the capture molecules may be accomplished by suitable solutions, e.g. appropriate buffer solutions are repealed.
  • suitable solutions e.g. appropriate buffer solutions are repealed.
  • the filter element can be rinsed both with the original filter flow and in the opposite direction.
  • other buffer solutions that allow the specific binding of the desired
  • the term substrate is understood in the following to mean a carrier material in which the individual constituents of the microfluidic filter element are arranged.
  • the microfluidic filter element has at least one filter chamber arranged in the substrate, which forms a cavity in which the actual filter material is arranged. This filter material in turn is with the immobilized capture molecules to bind the desired
  • microfluidic filter element this is composed of a plurality of superimposed substrates in which at least one filter chamber and the channels for supplying the biological sample fluid and for deriving the remaining after the filtering process
  • Filter material in the filter chamber can be flowed transversely over a large area.
  • the multi-layer substrate structure is also advantageous in the production, since the individual chambers and channel components can be better matched by their arrangement in different layers.
  • the microfluidic filter element is constructed from a substrate having at least one filter chamber and channels for supplying the biological sample fluid and for deriving the remaining components of the biological sample fluid after the filtering process, and the filter material is flowed laterally.
  • the microfluidic filter element has at least one further channel, at least one valve and / or at least one collecting chamber for the remaining components of the biological sample fluid.
  • the further channels are used, for example, for rinsing with buffer solutions or for eluting the bound sample components.
  • the valves regulate the flow rates within the channels.
  • individual or all channels can be equipped separately with valves. Since the filter effect does not depend crucially on the flow rate, it is possible for generating the flow use cheap, integrated pumps, such as peristaltic pumps that produce, for example, no continuous flow. Alternatively, however, are also external, ie outside the
  • Microfluidic system mounted, pumps for controlling the flow rate possible. Furthermore, within the substrate one or more collection chambers for the remaining components of the biological
  • the microfluidic filter element has channels with a height and a width of in each case 5 ⁇ m to 5 mm, preferably 50 ⁇ m to 2 mm, and the filter element is altogether for receiving a biological sample fluid having a volume of 1 ⁇ m to 20 ml , preferably 10 ⁇ to 10 ml.
  • the capture molecules are selected from the group of proteins, preferably antibodies or enzymes, polynucleic acids, preferably DNA or RNA, and polymers.
  • proteins preferably antibodies or enzymes, polynucleic acids, preferably DNA or RNA, and polymers.
  • catcher molecules are possible.
  • the catcher molecules are, for example, by silanization,
  • APTES Aminopropyltriethoxysilane
  • the catcher molecules are in each case specific to the sought
  • Sample components are matched and enter into a reversible or irreversible binding with these.
  • a direct implementation of the sought sample components by the capture molecules is possible. This is the case, for example, when the capture molecule is an enzyme which filters a particular enzyme substrate from the sample fluid.
  • Immobilization of the catcher molecules is carried out according to common methods such as Example of silanization.
  • the filter material is complete with the
  • Filter material selected from the group of fibers, preferably silica fibers; Membranes, preferably capillary membranes of a polymer material or cellulose; Beds of particles, preferably of colloids, more preferably of silica, polymethyl methacrylate or melamine; and metal meshes, preferably transmission electron microscopy grids.
  • Filter material to be immobilized.
  • different methods are used depending on the filter material.
  • the surface available for the interaction must be as large as possible.
  • every material comes with controllable
  • beds of particles such as colloids for use in the filter element according to the invention are suitable.
  • the function of the colloids is to produce a filter with adjustable pore size.
  • the actual separation of the desired sample components continues to take place via the catcher molecules.
  • silica polymethyl methacrylate or melamine are used.
  • the filter material consists of metal nets or metal grids in the manner as they are for example for
  • TEM grids are commercially available and have defined hole sizes of approx.
  • the filter element several 100 ⁇ to about 10 ⁇ on.
  • the superimposition of several networks or grids is provided in a further embodiment of the filter element.
  • the particles and metal mesh are coated with gold or platinum.
  • the filter material has a thickness of 100 ⁇ to 10 mm, preferably 30 ⁇ to 7 mm, more preferably 50 ⁇ to 5 mm and a pore size of 0.5 to 1000 ⁇ , preferably 1 to 500 ⁇ , particularly preferably 5 to 300 ⁇ , on.
  • the minimum thickness of the filter material is dictated by the thickness of a single TEM grid.
  • the pores of the filter material are dimensioned so that components of the sample liquid that do not contain any of the sample components sought can pass the filter substantially. In the case of bacteria in whole blood, this results in e.g. a minimum pore size of several ⁇ . Larger pores reduce the fluid resistance. At the same time, however, the efficiency of the filter material is lowered. This results depending on the application
  • the size of the filter chamber is adjusted in advance to the desired filter material and its extent. However, the diameter of the filter chamber is generally between 1 and 30 mm.
  • the substrate consists of a polymer, preferably a polycarbonate,
  • the substrate has a thickness of 0.2 to 8 mm, preferably 0.5 to 3 mm and an outer dimension of 10 x 10 to 100 x 100 mm 2 . Again, the dimensions of the substrate must be adapted to use in a microfluidic system.
  • microfluidic filter element takes place in a lab-on-chip system or in a purification tube.
  • Sample volumes (microliters) facilitated.
  • the application not only relates to the field of classical microfluidics, but also to other applications in which such a purification is to be achieved, e.g. by means of purification tubes.
  • Such art-known kit systems based on purification tubes are often used in in vitro diagnostics.
  • Fig. 1 shows schematically the filter principle of the filter element according to the invention
  • Fig. 2 shows the filter element according to the invention in plan view
  • FIG. 3 shows the microfluidic filter element as a layer system comprising a plurality of substrates 1
  • Fig. 1 shows schematically the filter principle of the filter element according to the invention using the example of a fiber filter.
  • catcher molecules 13 are immobilized.
  • the particular sample components 14 are separated by binding from the sample fluid.
  • Other sample components 15 pass through the filter material 12 due to the large pores in the filter material 12 substantially unhindered.
  • the figure is based on the example of a 13-functionalized fiber filter. In doing so, a biological sample fluid, e.g. Blood, bacteria 14 filtered.
  • the filter material 12 can pass largely unhindered. However, the principle can be applied to a variety of applications.
  • Fig. 2 shows a simple realization of the filter element according to the invention.
  • the channels for supplying the biological sample fluid 2 and for deriving the remaining after the filtering process components of the biological sample fluid 3, more channels 4/5 for the supply and removal of the buffer solution, as well as in the channels 21 3, 4/5 placed valves 6 / II 8/9 arranged.
  • the substrate 1 has a filter chamber 1 1 with therein befindlichem filter material 12 and a collecting chamber 10. With closed valves 8/9 sample fluid is introduced via channel 2 into the filter chamber 1 1 and flows through the
  • Filter material 12 laterally. While the specific sample components 14 bind to the respective capture molecules 13, the remaining
  • Sample liquid from the filter material 12 to remove can with a appropriate buffer solution rinsed.
  • the valves 8/9 are opened and the valves 6/7 are closed so that the channels 4/5 are opened.
  • the other sample components 15 can now be removed.
  • the sought-after specific sample components 14 can be further
  • the concentrated sample components 14 may be placed directly on the filter material 12 for analysis and diagnostics, e.g. for a PCR analysis, continue to be used.
  • 3 shows the microfluidic filter element as a layer system comprising a plurality of substrates 1. With this construction, a transverse flow through the filter material 12 becomes possible.

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  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Dispersion Chemistry (AREA)
  • Analytical Chemistry (AREA)
  • Hematology (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Molecular Biology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Fluid Mechanics (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Sampling And Sample Adjustment (AREA)

Abstract

L'invention concerne un élément filtrant microfluidique permettant de séparer des constituants d'un échantillon de fluide biologique. Ledit élément filtrant comporte au moins un substrat, divers canaux, au moins une chambre de filtration agencée dans le substrat et dotée d'un matériau filtrant, ainsi que des molécules de fixation immobilisées sur le matériau filtrant et pénétrant dans le matériau filtrant.
PCT/EP2012/068901 2011-11-14 2012-09-26 Élément filtrant microfluidique permettant de séparer des constituants d'un échantillon de fluide biologique WO2013072110A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE201110086235 DE102011086235A1 (de) 2011-11-14 2011-11-14 Mikrofluidisches Filterelement zum Abscheiden von Probenbestandteilen aus einem biologischen Probenfluid
DE102011086235.8 2011-11-14

Publications (1)

Publication Number Publication Date
WO2013072110A1 true WO2013072110A1 (fr) 2013-05-23

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Application Number Title Priority Date Filing Date
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Country Link
DE (1) DE102011086235A1 (fr)
WO (1) WO2013072110A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2905334A1 (fr) * 2014-02-11 2015-08-12 Robert Bosch Gmbh Système d'enzyme, kit et procédé d'exécution de réaction enzymatiques
US20170088880A1 (en) * 2014-05-15 2017-03-30 Robert Bosch Gmbh Device and Method for Processing a Biological Sample and Analysis System for Analyzing a Biological Specimen
US11311879B2 (en) * 2018-02-05 2022-04-26 miDiagnostics NV Microfluidic device, system, and method for reversing a flow through a microfluidic channel

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2019514017A (ja) 2016-04-28 2019-05-30 ヒューレット−パッカード デベロップメント カンパニー エル．ピー．Ｈｅｗｌｅｔｔ‐Ｐａｃｋａｒｄ Ｄｅｖｅｌｏｐｍｅｎｔ Ｃｏｍｐａｎｙ， Ｌ．Ｐ． マイクロ流体濾過
GB2560379A (en) * 2017-03-10 2018-09-12 Epigem Ltd Microfluidic device
DE102019215031A1 (de) * 2019-09-30 2021-04-01 Robert Bosch Gmbh Einrichtung und Verfahren zur Überführung einer Probe, insbesondere einer Blutprobe, in eine mikrofluidische Vorrichtung

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2401942A (en) 2003-05-22 2004-11-24 Bioct 5 Ltd Polypeptide and polynucleotide assay methods and apparatus
EP2070594A1 (fr) * 2007-12-14 2009-06-17 Koninklijke Philips Electronics N.V. Dispositif microfluidique et son procédé de fabrication et capteur l'incorporant
WO2009150583A1 (fr) * 2008-06-10 2009-12-17 Koninklijke Philips Electronics N.V. Dispositif de diagnostic
WO2011038458A1 (fr) * 2009-09-30 2011-04-07 Mycrolab Diagnostics Pty Ltd Réduction sélective d'adhérence dans des dispositifs microfluidiques
US20110129931A1 (en) * 2009-10-20 2011-06-02 Agency For Science, Technology And Research Microfluidic system for detecting a biological entity in a sample

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2401942A (en) 2003-05-22 2004-11-24 Bioct 5 Ltd Polypeptide and polynucleotide assay methods and apparatus
EP2070594A1 (fr) * 2007-12-14 2009-06-17 Koninklijke Philips Electronics N.V. Dispositif microfluidique et son procédé de fabrication et capteur l'incorporant
WO2009150583A1 (fr) * 2008-06-10 2009-12-17 Koninklijke Philips Electronics N.V. Dispositif de diagnostic
WO2011038458A1 (fr) * 2009-09-30 2011-04-07 Mycrolab Diagnostics Pty Ltd Réduction sélective d'adhérence dans des dispositifs microfluidiques
US20110129931A1 (en) * 2009-10-20 2011-06-02 Agency For Science, Technology And Research Microfluidic system for detecting a biological entity in a sample

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2905334A1 (fr) * 2014-02-11 2015-08-12 Robert Bosch Gmbh Système d'enzyme, kit et procédé d'exécution de réaction enzymatiques
US20170088880A1 (en) * 2014-05-15 2017-03-30 Robert Bosch Gmbh Device and Method for Processing a Biological Sample and Analysis System for Analyzing a Biological Specimen
US10308977B2 (en) * 2014-05-15 2019-06-04 Robert Bosch Gmbh Device and method for processing a biological sample and analysis system for analyzing a biological specimen
US11311879B2 (en) * 2018-02-05 2022-04-26 miDiagnostics NV Microfluidic device, system, and method for reversing a flow through a microfluidic channel
US20220226821A1 (en) * 2018-02-05 2022-07-21 miDiagnostics NV Microfluidic device, system, and method for reversing a flow through a microfluidic channel
US11590500B2 (en) 2018-02-05 2023-02-28 miDiagnostics NV Microfluidic device, system, and method for reversing a flow through a microfluidic channel

Also Published As

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