WO2003011768A2 - Microfluidic device for molecular analysis - Google Patents
Microfluidic device for molecular analysis Download PDFInfo
- Publication number
- WO2003011768A2 WO2003011768A2 PCT/US2002/024316 US0224316W WO03011768A2 WO 2003011768 A2 WO2003011768 A2 WO 2003011768A2 US 0224316 W US0224316 W US 0224316W WO 03011768 A2 WO03011768 A2 WO 03011768A2
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- molecules
- molecule
- signal delivery
- sample
- chamber
- Prior art date
Links
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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6813—Hybridisation assays
- C12Q1/6816—Hybridisation assays characterised by the detection means
-
- 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
- B01L2200/10—Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
-
- 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
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2400/00—Moving or stopping fluids
- B01L2400/04—Moving fluids with specific forces or mechanical means
- B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
- B01L2400/0415—Moving fluids with specific forces or mechanical means specific forces electrical forces, e.g. electrokinetic
- B01L2400/0421—Moving fluids with specific forces or mechanical means specific forces electrical forces, e.g. electrokinetic electrophoretic flow
Definitions
- the present invention relates to the analysis of target solutions utilizing
- microelectromechanical systems including microfluidic devices and methods.
- the present invention relates to microelectromechanical systems (MEMS), and particularly
- microfluidic devices for the analysis of biochemical or chemical components.
- biochips known as biochips
- microfluidic devices 6,284,113 (Apparatus and method for transferring liquids); 6,176,962 (Apparatus and method for transferring liquids); 6,176,962
- Gamera's work is disclosed in 6, 143,248 (Capillary microvalve) and 6,063,589 (Devices and
- the chamber being provided for preparation of the sample via pulsed field lysis /
- biochip to further channels (or chambers) using applied electric fields.
- the applied fields are
- microfluidic devices having a
- the signal delivery molecules being
- signal delivery molecules further having a signalling component.
- microfluidic devices having a
- microfluidic devices comprising
- the signalling component the signal delivery molecules being branched nucleic acids.
- said complex comprises a
- the signal delivery molecule preferably being a branched
- nucleic acid e.g. a 3DNA dendrimer having the desired signal molecules linked thereto.
- a microfluidic device comprising a signal delivery chamber, the signal
- branched nucleic acid signal delivery molecules having a binding site with an affinity for
- an improved microfluidic device for the detection and analysis of desired chemical or biochemical components.
- microfluidic devices allow molecular biological analysis and diagnosis using multistep reactions
- components provided in the biochip utilize suitable techniques from molecular biology in a
- This circuitry preferably includes electronic contacts
- internal chip electrodes provide desired AC
- the present systems can be used for medical use
- the device in the preferred embodiment of the invention, the device
- the device is
- telomerase associated RNA and telomerase mRNA used for the detection and measurement of telomerase associated RNA and telomerase mRNA
- microfluidic device is provided with
- an initial sample area such as a chamber where whole cells are added, with the cells being
- sample in the sample chamber are transported through a size specific filter or plastic frit to a
- Target molecules by electrophoretic mobility. Molecules of the desired mobility are transferred
- the signal delivery molecule is
- branched nucleic acid which has been constructed by hybridization and/or cross-
- the signal delivery molecules are dendrimer molecules (e.g.
- the complex is then transferred through a further channel to an analysis
- the target molecules are detected at an electrode. It is further preferred that the target molecules are detected at an electrode.
- Figure 1 is a schematic of a microfluidic biochip device in accordance with one preferred
- Figure 2 is a schematic of a microfluidic biochip device in accordance with a further embodiment
- biochip device is provided as shown, for
- Biochip device 12 includes a series of sample wells and channels therein
- processed discussed as conducted in a chamber can be conducted in a channel (or vice versa),
- the biochip itself is constructed of any material or surface suitable for the particular application
- plastic preferably plastic, glass or silicon.
- silicon Alternatively, any other desired materials can be
- the internal structure of the biochip is created by molding the chip of two separate
- the halves one or both of the halves having the desired channel and well structure.
- the halves having the desired channel and well structure.
- bottom half is molded with the desired structure, and the top is provided as a flat coverslip
- the chip is incorporated within the structure of the chip.
- the chip is plastic.
- Biochip device 12 includes a sample addition well 26 where whole sample cells or a
- sample lysate are added to the device for analysis.
- the sample can be added to the well 26
- a preferred cell population to load into the sample well is
- telomerase positive circulating cancer cells prepared by the Hexal/Gentech OncoquickTM device. Upon addition of the whole cells to sample well or chamber 26, the cells are lysed within
- the cells are lysed using pulsed field sample cell disruption. Additionally or
- the cells are added in an osmotically positive buffer relative to the cells.
- the cells can be lysed using lysing reagents.
- sample addition chamber 26 is approximately is 5ul to
- the chamber 26 is provided with
- the x axis is
- Sample disruption electrodes 18 are used to apply joule heating and disruption of the
- the lysis can be conducted with or without sharpened electrodes
- biochip can likewise be used to detect positively charged molecules by reversing the polarity of
- the target molecule or substance can
- nucleic acids any charged natural or artificial substance, including, but are not limited to, nucleic acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids
- proteins proteins, peptide, carbohydrates, lipids, polysaccarides, glycoproteins, hormones, receptors,
- antigens antibodies, viruses, pathogens, metabolic byproducts, growth factor, cofactors,
- Collection channel 30 is a crude analyte collection
- target molecules from fragments of cell membrane, cell debris, and other undesirable
- the collection channel contains a plastic frit or other filter.
- This initial filter is preferably provided with a nominal porosity of 50nm to 200 microns, or of
- each of the anodes and cathodes of the biochip are identical to each of the anodes and cathodes of the biochip.
- electrodes can be provided in the channels.
- the channels can be provided in the channels.
- electrodes can be located in each chamber or channel behind a frit which has a porosity smaller
- the electrodes can be separate from the chamber or
- the electrodes are shaped and/or located to produce an
- a parabolic electrode is provided for each anode and cathode so as
- the analyte molecules are electrically focused to the middle of the second
- electrophoretic separation channel 40 electrophoretic separation channel 40. Furthermore, in the preferred embodiment
- second chamber is of such duration and intensity to ensure that only relatively "small" molecules
- RNA molecules reach the crude second sample chamber 36, thereby excluding "large"
- chromosomal nucleic acids molecules such as chromosomal nucleic acids (with small being typically up to approximately
- intensity and duration are applied to anode 32 and cathode 42 to establish an electric field which directs the target molecules into the second channel, electrophoretic separation channel 40.
- Channel 40 is designed to size fractionate the negatively charged molecules located in the crude
- Channel 40 is preferably filled with polyacrylamide
- channel 40 (typically 4-12 w/v%) for electrophoresis.
- the behavior of channel 40 is utilized to fractionate
- the device uses a first
- a delivery or detection channel 50 is located at the location where
- the target molecule is electrophoretically focused, the channel 50 being provided for subsequent
- the channel can be provided
- electrophoresis in separation channel 40 (appropriate being defined such that the center of the
- concentration gradient of analyte molecules is centered on the delivery channel 50), the y-axis
- RNA target analyte molecules
- the hybrization well also referred to herein as the signal delivery well
- Hybridization or signal delivery well 46 is a chamber provided with a relatively large
- the signal delivery molecules are molecules having a binding area with an affinity for
- the desired target molecule of interest and having a signalling component, the signalling
- the signal delivery molecules are branched nucleic acids, the nucleic acids being branched by hybridization and/or
- the signal delivery molecules are dendrimers (e.g. 3DNA)
- Dendrimer technology is
- the dendrimer molecule specificity is conferred via any analyte-recognizing-moiety
- the specificity is preferably
- DNA DNA, RNA, PNA, LNA or any polymer that specifically recognizes via base pairing with the
- the signal delivery molecules are preferably predispensed into the third chamber prior to
- appropriate microvalves are provided at the entrance and
- the signal delivery molecule can be dispensed into chamber 46 by
- the target molecules delivered through the hybridization or signal delivery well 46 the target molecules delivered through
- the detection channel 50 bind to the predetermined signal delivery molecules (e.g. dendrimer
- the signal delivery molecule (preferably a branched nucleic acid or a hyperbranched nucleic acid such as a dendrimer) contain the signalling component(s) which
- the signal is an electrical current generated at the working electrode (the detection electrode).
- the signalling component or signal molecule of the signal is selected from the group consisting of:
- delivery molecules are the "G" residues of a 3DNA molecule, as previously disclosed by the H.
- another preselected base e.g., adenine, 6-mercaptoguanine, 8-oxo-guanine, or 8-oxo-adenine.
- the nucleic acid is reacted with a transition metal complex capable
- reaction is detected to determine the presence or absence of the nucleic acid from the detected
- the signal molecules are electron
- suitable electron donors include the
- rare earth cryptands and caged neutral metal atoms, and coordinated metals that can be further
- a preselected base and electron donor molecules are both utilized for signalling.
- the target molecules Upon delivery of the target molecule to chamber 46, the target molecules bind to the
- voltages are applied in the y-axis to electrodes 72 and 82 to
- a working electrode 90 (the “detection electrode") is provided in analysis chamber 56.
- the binding preferably at the end of the signal delivery molecule (or alternatively that will bind the signal delivery molecule), the binding preferably
- neutral capture molecules are utilized.
- DNA oligonucleotides can be used at the working
- capture molecules are provided which bind to a site on
- each target molecule which is different from the target's binding site for the signal delivery
- the target molecule serve as a bridge between the
- working electrode that serve to capture the target molecule are electrochemically neutral.
- the capture molecules In the preferred embodiment and for nucleic acid (RNA) targets, the capture molecules
- RNA preferably composed of PNA, DNA, RNA, LNA or any polymer that specifically recognizes
- these capture molecules lack the "G” base so as not to contribute to the subsequent
- a voltage is applied in the x axis to electrodes 92 and 102 to cause unbound
- axis can be tuned to the strength of the interaction between the target molecule and signal
- stringency the present application refers to the ability to
- hybridizations of the present invention preferably an electric stringency is utilized.
- the signal molecules on the signal delivery molecules e.g. 3DNA
- nucleic acids are indium doped tin oxide modified with 4-vinyl-4'-methyl-2,2'-bipyridine or
- the total amount of current generated at the working electrode is then measured by the
- nucleic acid hybrization analysis or in place of, nucleic acid hybrization analysis, antibody/antigen analysis can be used or so forth
- more than one target molecule can be any one target molecule.
- separation channel 40 As shown in Figure 1, in a first embodiment as
- desired target molecules are subsequently migrated through a delivery or
- a second group of molecules can be isolated and delivered through a second delivery
- fourth chambers 156, 256, and fifth chambers 166, 266 (and associated channels) are provided
- the biochip can be used to simultaneously separate both negatively and positively
- a second network of chambers is provided
- the network provided on the left of the first chamber operates using electrodes and voltages of opposite polarities to those
- Appendix B Aclara References
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- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Analytical Chemistry (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Immunology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Physics & Mathematics (AREA)
- Genetics & Genomics (AREA)
- Dispersion Chemistry (AREA)
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Abstract
Description
Claims
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US30922101P | 2001-08-01 | 2001-08-01 | |
US60/309,221 | 2001-08-01 |
Publications (2)
Publication Number | Publication Date |
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WO2003011768A2 true WO2003011768A2 (en) | 2003-02-13 |
WO2003011768A3 WO2003011768A3 (en) | 2003-05-30 |
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PCT/US2002/024316 WO2003011768A2 (en) | 2001-08-01 | 2002-08-01 | Microfluidic device for molecular analysis |
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Cited By (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE10353985A1 (en) * | 2003-11-19 | 2005-06-23 | Olympus Biosystems Gmbh | Apparatus for manipulation and analysis of micro-objects, useful particularly for cells or their components, is constructed as a fluidics microsystem and/or microchip |
GB2416030A (en) * | 2004-01-28 | 2006-01-11 | Norchip As | A diagnostic system for carrying out a nucleic acid sequence amplification and detection process |
EP1672059A1 (en) * | 2004-12-17 | 2006-06-21 | Samsung Electronics Co., Ltd. | Microfluidic device comprising electrolysis device for cell lysis and method for electrochemically lysing cells |
WO2010141385A1 (en) * | 2009-06-01 | 2010-12-09 | Geron Corporation | Assay for telomerase activity using microfluidic device |
DE102010064391A1 (en) * | 2010-10-29 | 2012-05-03 | Endress + Hauser Conducta Gesellschaft für Mess- und Regeltechnik mbH + Co. KG | Method for determining an analyte content of a liquid sample by means of a bioanalyzer |
WO2012064704A1 (en) * | 2010-11-08 | 2012-05-18 | Daktari Diagnostics, Inc. | Multi-function microfluidic test kit |
CN104897756A (en) * | 2015-06-19 | 2015-09-09 | 青岛大学 | Electrochemical sensor for detecting telomerase activity and method for manufacturing electrochemical sensor |
WO2018050867A1 (en) * | 2016-09-15 | 2018-03-22 | Dianax S.R.L. | Microfluidic device, particularly of the lab-on-chip type, for the concentration, purification, and detection of biological and/or medical targets of interest from a biological sample |
CN108136394A (en) * | 2015-10-05 | 2018-06-08 | 豪夫迈·罗氏有限公司 | For the kit of cell detection |
US10046321B2 (en) * | 2013-09-19 | 2018-08-14 | Dianax S.R.L. | Diagnostic device, particularly of the lab-on-chip type |
WO2023223331A1 (en) * | 2022-05-18 | 2023-11-23 | The State Of Israel, Ministry Of Agriculture & Rural Development, Agricultural Research Organization (Aro) (Volcani Institute) | Dual-mode electrochemical point-of-care detection, quantification and profiling of pathogens |
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US6406893B1 (en) * | 1997-04-04 | 2002-06-18 | Caliper Technologies Corp. | Microfluidic methods for non-thermal nucleic acid manipulations |
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Cited By (21)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE10353985A1 (en) * | 2003-11-19 | 2005-06-23 | Olympus Biosystems Gmbh | Apparatus for manipulation and analysis of micro-objects, useful particularly for cells or their components, is constructed as a fluidics microsystem and/or microchip |
GB2416030A (en) * | 2004-01-28 | 2006-01-11 | Norchip As | A diagnostic system for carrying out a nucleic acid sequence amplification and detection process |
GB2416030B (en) * | 2004-01-28 | 2008-07-23 | Norchip As | A diagnostic system for carrying out a nucleic acid sequence amplification and detection process |
EP1672059A1 (en) * | 2004-12-17 | 2006-06-21 | Samsung Electronics Co., Ltd. | Microfluidic device comprising electrolysis device for cell lysis and method for electrochemically lysing cells |
US7488596B2 (en) | 2004-12-17 | 2009-02-10 | Samsung Electronics Co., Ltd. | Microfluidic device comprising electrolysis device for cell lysis and method for electrochemically lysing cells using the same |
US7598064B2 (en) | 2004-12-17 | 2009-10-06 | Samsung Electronics Co., Ltd. | Microfluidic device comprising electrolysis device for cell lysis and method for electrochemically lysing cells using the same |
WO2010141385A1 (en) * | 2009-06-01 | 2010-12-09 | Geron Corporation | Assay for telomerase activity using microfluidic device |
US20120070842A1 (en) * | 2009-06-01 | 2012-03-22 | Harley Calvin B | Assay for Telomerase Activity Using Microfluidic Device |
US10036098B2 (en) | 2010-10-29 | 2018-07-31 | Endress+Hauser Conducta Gmbh+Co. Kg | Method for determining an analyte content of a liquid sample by means of a bioanalyzer |
DE102010064391A1 (en) * | 2010-10-29 | 2012-05-03 | Endress + Hauser Conducta Gesellschaft für Mess- und Regeltechnik mbH + Co. KG | Method for determining an analyte content of a liquid sample by means of a bioanalyzer |
DE102010064392A1 (en) * | 2010-10-29 | 2012-05-03 | Endress + Hauser Conducta Gesellschaft für Mess- und Regeltechnik mbH + Co. KG | Method for determining an analyte content of a liquid sample by means of a bioanalyzer |
WO2012064704A1 (en) * | 2010-11-08 | 2012-05-18 | Daktari Diagnostics, Inc. | Multi-function microfluidic test kit |
US10046321B2 (en) * | 2013-09-19 | 2018-08-14 | Dianax S.R.L. | Diagnostic device, particularly of the lab-on-chip type |
CN104897756B (en) * | 2015-06-19 | 2015-12-02 | 青岛大学 | A kind of electrochemical sensor for test side telomerase activity and preparation method thereof |
CN104897756A (en) * | 2015-06-19 | 2015-09-09 | 青岛大学 | Electrochemical sensor for detecting telomerase activity and method for manufacturing electrochemical sensor |
CN108136394A (en) * | 2015-10-05 | 2018-06-08 | 豪夫迈·罗氏有限公司 | For the kit of cell detection |
CN108136394B (en) * | 2015-10-05 | 2021-07-02 | 豪夫迈·罗氏有限公司 | Kit for cell detection |
WO2018050867A1 (en) * | 2016-09-15 | 2018-03-22 | Dianax S.R.L. | Microfluidic device, particularly of the lab-on-chip type, for the concentration, purification, and detection of biological and/or medical targets of interest from a biological sample |
WO2018051374A1 (en) * | 2016-09-15 | 2018-03-22 | Dianax S.R.L. | Microfluidic device, particularly of the lab-on-chip type, for the concentration, purification, and detection of biological and/or medical targets of interest from a biological sample |
US11229908B2 (en) | 2016-09-15 | 2022-01-25 | Dianax S.R.L. | Microfluidic device, particularly of the lab-on-chip type, for the concentration, purification, and detection of biological and/or medical targets of interest from a biological sample |
WO2023223331A1 (en) * | 2022-05-18 | 2023-11-23 | The State Of Israel, Ministry Of Agriculture & Rural Development, Agricultural Research Organization (Aro) (Volcani Institute) | Dual-mode electrochemical point-of-care detection, quantification and profiling of pathogens |
Also Published As
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