EP1894007A1 - Verfahren und vorrichtung zur quantitativen bestimmung von analyten in flüssigen proben - Google Patents
Verfahren und vorrichtung zur quantitativen bestimmung von analyten in flüssigen probenInfo
- Publication number
- EP1894007A1 EP1894007A1 EP06743076A EP06743076A EP1894007A1 EP 1894007 A1 EP1894007 A1 EP 1894007A1 EP 06743076 A EP06743076 A EP 06743076A EP 06743076 A EP06743076 A EP 06743076A EP 1894007 A1 EP1894007 A1 EP 1894007A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- carrier
- light
- sample
- substances
- analytes
- 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.)
- Ceased
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54366—Apparatus specially adapted for solid-phase testing
- G01N33/54373—Apparatus specially adapted for solid-phase testing involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y10/00—Nanotechnology for information processing, storage or transmission, e.g. quantum computing or single electron logic
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
Definitions
- the present invention relates to a method and a device for the highly sensitive parallel detection of analytes in liquid media.
- the object of the present invention is therefore to provide a cost-effective method and a device for fast, highly sensitive and parallel detection of a plurality of different analytes.
- first layers are applied to a carrier surface.
- the carrier material used is a light-conducting medium.
- this glass or plastic is suitable.
- the carrier has two parallel surfaces in a particularly preferred embodiment.
- a polymer layer is applied first according to the method as described in DE19816604 A1.
- a layer is applied which has substances with molecular structures which correspond to or are similar to the analyte to be investigated.
- the molecular structures of the applied substances have suitable properties for sufficiently specific detection by the recognition structures of the ligands added later in the binding test of the sample.
- the applied substances may correspond to the analyte or a derivative of the analyte to be examined, which have suitable functional groups in order to be covalently or noncovalently bound to the polymer layer. If only one analyte is to be detected, the corresponding molecular structures are applied in a planar manner. In the case of several analytes to be examined, the application of different molecular structures takes place in a spatially resolved manner, e.g. in the form of spots.
- the samples to be examined are prepared for the binding inhibition test.
- the samples are diluted, if necessary, with a suitable liquid.
- the analytes are transferred to a liquid medium using suitable methods (extraction, solution, grinding, or the like).
- defined volumes of the liquid samples are pre-incubated with the appropriate ligands.
- ligands substances are understood here and below which have suitable recognition structures for sufficiently specific detection and binding of analytes to be examined from the sample.
- antibodies, aptamers, antigens, coated Beads etc. are used.
- the ligands bear suitable markers for later detection, which can be excited by an evanescent field. These may be, for example, fluorescent dyes, quantum dots or the like.
- the preincubation of the sample is terminated either after reaching equilibrium or after expiration of a predefined time.
- the pretreated sample is brought into contact with the coated support.
- a flow method is used.
- the sample is passed through a flow cell in or on which the coated carrier is located, so that the sample comes into contact with the coated surface.
- the ligands in the sample can bind via the free binding sites to the corresponding molecular structures of the substances applied to the support surface.
- the incubation can be stopped by rinsing the carrier.
- the laser light is used, which is generated for example by light-emitting diodes (LED), laser diodes or laser.
- LED light-emitting diodes
- laser diodes laser diodes
- an evanescent field arises at the phase boundary, which excites the markers located on the surface of the support.
- the marker used and the light used must be coordinated so that an excitation can take place.
- the marker emits its characteristic light, which is detected by a detector.
- photodiodes or CCD elements can be used as the detector.
- appropriate filters can be placed between the carrier and the detector.
- additional coupling and decoupling elements for example in the form of mirrors, lenses or light guides, can be arranged between the light source and the carrier and / or between the carrier and the detector.
- the light intensities measured by the detector can now be evaluated and used as a basis for the quantitative determination of the analytes to be examined.
- an evaluation device in particular a computer can be used, so that the evaluation is automated.
- further method steps such as sample preparation and the measurement process can be carried out according to the invention of machines. In a particularly preferred embodiment, the entire process is fully automatic.
- analytes such as hormones, antibiotics, pesticides, pharmaceuticals, drugs and other molecules or molecular complexes
- Different types of fluids can be analyzed, such as drinking water, fruit juices, milk, serum, blood plasma, urine and others.
- ligands with suitable recognition structures for the respective analytes, substances for the carrier surface with suitable molecular structures, and suitable sample preparation are selected.
- the flexibility of the method allows its application in different areas: from food monitoring to water analysis to clinical diagnostics.
- the method according to the invention allows several diverse analytes to be detected simultaneously in different liquid media more quickly, more sensitively and more cost-effectively.
- FIG. 1 The principle of the binding inhibition test used in the method according to the invention using the example of an immunoassay; the antibodies 1 serve as ligands for the analyte 2; A: In the first step, pre-incubation is performed, in which antibody 1 is added to the sample containing the analyte 2; B: In the second step, the sample is pumped over the carrier 4; the antibodies 1 can now bind with their free binding sites to the modified surface 3; after completion of the incubation, the detection takes place.
- the light is coupled from the light source 1 in the glass substrate 2 and forwarded by total reflection within the carrier 2; on the carrier surface, which is coated with corresponding substances 3 for the specific detection of the analyte, an evanescent field is formed near the surface; the sample can be brought into contact with the carrier surface via a flow cell 4; after the incubation of the prepared sample, the detection of the emitted light is carried out by a detector 5; this transmits the recorded measurement data to an evaluation unit 6.
- Fig. 3 Calibration curve for progesterone in UHT milk; Progesterone concentrations between 0.009 and 900 ng ml '1 were measured (six steps); the antibody was used at a concentration of 30 ng ml -1 in each sample and a detection limit of 46 pg ml -1 could be achieved.
- Fig. 4 Calibration curve for progesterone in fresh milk; progesterone concentrations were measured between 0.009 to 900 ng ml -1 (six steps) and the antibody was used at a concentration of 30 ng ml -1 in each sample; a detection limit of 56 pg ml-1 could be achieved.
- Fig. 5 Calibration curve for progesterone in raw milk; progesterone concentrations were measured between 0.009 to 900 ng ml -1 (six steps) and the antibody was used at a concentration of 30 ng ml -1 in each sample; a detection limit of 52 pg ml -1 was achieved.
- Fig. 6 Calibration curve for testosterone in bovine serum; testosterone concentrations between 0.009 and 900 ng ml -1 were measured (six steps) and the antibody was used at a concentration of 30 ng ml -1 in each sample; a detection limit of 309 pg ml -1 was achieved.
- the hormone progesterone in three different types of milk is quantified. Detection limits between 46 and 56 pg ml "1 were achieved.
- Consumption chemicals were purchased from Sigma-Aldrich and Merck KGaA.
- the hormone was purchased as VETRANAL ® standard for Riedl-de Haen Laboratory Chemicals GmbH & Co. KG.
- the monoclonal IgGI antibody, anti-progesterone, was purchased from Achs Antibodies GmbH.
- the fluorescence marker CyDye Cy5.5 used was purchased from Amersham Biosciences Europe GmbH.
- the aminodextran Amdex TM 40,000 dalton molecular weight was purchased from the Helix Research Company.
- the progesterone derivative for immobilization on the support surface was synthesized.
- the basic unit consists of a 1 ml reciprocating syringe with T-valve for the Tecan Cavro-Module XL3000; a sample loop consisting of a teflon tube with about 2 ml total volume of Ismatec; a 6-way valve with a flow cell tube (0.7 ml total volume) from Ismatec; a Plexiglas flow cell with milled flow channel and Swagelok ports for inlet and outlet of proliquid; a Bok7 glass size 60x14x1.5mm bulkhead glass from Desag, with the 45 ° bevel and polish made by PE Applied Biosystems; a modulated laser diode with a wavelength of 635 nm and 15 mW power from Cohenent; six polymer fibers with a numerical aperture of 0.46; six edge filters 680 AELP with a diameter of 25 mm, a thickness of 4.5 mm and a maximum transmission of 90% from Omega Optical; six photodiodes with integrated preamplifier
- the autosampler HTS PAL by CTC Analytics was used for sample preparation. It consists mainly of a moving 1 ml syringe, a washing station, a six-port Inject / Load valve (Valco) Sample holder for 98 1 ml samples and one sample holder for five 10 ml samples.
- the software program Cycle Composer controls the autosampler via a separate PC. The communication between the two PCs takes place via a relay card.
- the autosampler automatically mixes the samples and injects them into the own sample loop via the Valco valve (Teflon tube with 960 ⁇ l, Ismatek).
- the carrier is completely coated, whereas in the multi-analyte measurements a spatially resolved modification of the carriers using a microdosing system is required.
- the glass surface is first cleaned and activated.
- the glass slides are placed in a fresh piranha solution for 30 minutes and then rinsed well with deionized water.
- the carrier is covered with 50 .mu.l of GOPTS, a second placed on it (sandwich technique) and both stored in a dry chamber.
- the carriers are rapidly rinsed with dry acetone and dried in a stream of nitrogen.
- the activated carriers are covered with 50 ⁇ l of aminodextran-water solution and folded together (sandwich technique). Overnight they are stored in a steam atmosphere. Then they are rinsed with deionized water and dried. Now follows the reaction with the derivative. For this purpose, about 5 mg of the derivative is dissolved in a little dry DMF and mixed with 1, 1-fold molar amount of DCC in DMF. This solution is placed on the carrier and stored again with the sandwich technique in a DMF saturated chamber for at least 5 h. Thereafter, the carriers are rinsed first with DMF and then with deionized water.
- the GOPTS-activated carrier is dripped with a conjugate of derivative and aminodextran using the Microdrop dosing system.
- the spot diameter is 3 mm and the distance of the spots 6.5 mm.
- the conjugate is prepared from the active ester of the derivative and antimodextran (40 kD).
- the active ester of the derivative and antimodextran 40 kD.
- the aminodextran is dissolved in a mixture of carbonate buffer pH 9.5 and DMF (1: 1). 0.125 molar equivalents of active ester are added to the AMD solution.
- the solution is shaken overnight. Subsequently, the conjugate is precipitated with methanol, washed and freeze-dried. The conjugate is dissolved in deionized and filtered water (2 mg ml -1 ), which is used to drip the carrier by microdosing.
- the optical design of the device consists of a laser diode, which has a distance of approx. 2-5 cm from the carrier. Over the beveled edge of the glass carrier, the laser light is coupled into this. Through total reflection, the beam is transmitted within the carrier. The reflection points are at a distance of about 6.5 mm. At these sites, an evanescent field is formed near the surface in the flow cell, in which fluorescent dyes can be excited. On the back of the carrier polymer fibers conduct the fluorescence via edge filters to the photodiodes. The lock-in technique used modulates the laser light, and only the incoming, correspondingly modulated radiation is detected.
- the sample is mixed by the HTS PAL Autosampler and injected into the sample loop of the Valcoventils.
- the syringe pump then slowly pumps the sample over the flow cell.
- the antibody will be added to the sample immediately before the measurement. This has several advantages: First, the time interval between mixing and measurement is always the same, second, the antibody is spared, since it is only relatively short time in possibly aggressive matrices, and third, the antibody can be stored refrigerated in the storage vessel, without the complete sample tray needs to be cooled.
- the hormone testosterone is quantified in bovine serum.
- the hormone testosterone was purchased as VETRANAL ® Standard from Riedl-de Haen Laboratory Chemicals GmbH & Co. KG.
- the monoclonal IgGI antibody, anti-testosterone, was purchased from Achs Antibodies GmbH.
- the fluorescent marker CyDye TM Cy5.5 was purchased from Amersham Biosciences Europe GmbH.
- the 40,000 dalton molecular weight aminodextran Amdex TM was purchased from HeNx Research Company.
- the testosterone derivative (testosterone 3- (O-carboxymethyl) oxime) for immobilization on the support surface was purchased from Sigma-Aldrich.
- Example 1 The basic device and the optical design of the device correspond to the system used in Example 1.
- the HTS PAL autosampler from CTC Analytics was used according to the procedure described in Example 1.
- the complete coating method described in Example 1 was used for a single analyte measurement. Sample preparation and measurement procedure, as well as the calculation of the analytical parameters were carried out according to Example 1.
Landscapes
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Nanotechnology (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Hematology (AREA)
- Crystallography & Structural Chemistry (AREA)
- Urology & Nephrology (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Food Science & Technology (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biophysics (AREA)
- General Engineering & Computer Science (AREA)
- Medical Informatics (AREA)
- Analytical Chemistry (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Mathematical Physics (AREA)
- Theoretical Computer Science (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
Description
Claims
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE102005026839A DE102005026839A1 (de) | 2005-06-10 | 2005-06-10 | Verfahren und Vorrichtung zur quantitativen Bestimmung von Analyten in flüssigen Proben |
PCT/EP2006/005078 WO2006131225A1 (de) | 2005-06-10 | 2006-05-26 | Verfahren und vorrichtung zur quantitativen bestimmung von analyten in flüssigen proben |
Publications (1)
Publication Number | Publication Date |
---|---|
EP1894007A1 true EP1894007A1 (de) | 2008-03-05 |
Family
ID=36809093
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP06743076A Ceased EP1894007A1 (de) | 2005-06-10 | 2006-05-26 | Verfahren und vorrichtung zur quantitativen bestimmung von analyten in flüssigen proben |
Country Status (5)
Country | Link |
---|---|
US (1) | US20100297671A1 (de) |
EP (1) | EP1894007A1 (de) |
DE (1) | DE102005026839A1 (de) |
NZ (1) | NZ564074A (de) |
WO (1) | WO2006131225A1 (de) |
Families Citing this family (16)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP2167942B1 (de) | 2007-07-12 | 2021-05-19 | ASMAG-Holding GmbH | Optoelektronisches sensorsystem |
DE102009019476A1 (de) | 2009-05-04 | 2010-11-11 | Biametrics Marken Und Rechte Gmbh | Wiedererkennbarer Träger für optische Meßverfahren |
DE102009019711A1 (de) | 2009-05-05 | 2010-11-18 | Biametrics Marken Und Rechte Gmbh | Verfahren und Vorrichtung zur Bestimmung von Reflexionskoeffizienten an Filteranordnung mit dünnen Schichten |
DE102010041426A1 (de) | 2010-09-27 | 2012-05-03 | Siemens Aktiengesellschaft | Messeinheit und Verfahren zur optischen Untersuchung einer Flüssigkeit zur Bestimmung einer Analyt-Konzentration |
CN103354902B (zh) | 2010-12-06 | 2016-03-02 | 特拉维夫大学拉玛特有限公司 | 药物检测方法和试剂盒 |
JP2014508921A (ja) | 2011-01-31 | 2014-04-10 | ビアメトリクス ゲゼルシャフト ミット ベシュレンクテル ハフツング | 複数の波長の光を用いて薄膜層における強度を同時に測定することによって光学特性を決定する方法及び装置 |
GB2495703A (en) * | 2011-10-12 | 2013-04-24 | Crowcon Detection Instr Ltd | Optical sensor without wavelength filter |
DE102011085473A1 (de) | 2011-10-28 | 2013-05-02 | Albert-Ludwigs-Universität Freiburg | Verfahren zur Identifikation von Aptameren |
US9678015B2 (en) | 2014-09-26 | 2017-06-13 | Frito-Lay North America, Inc. | Method for elemental analysis of a snack food product in a dynamic production line |
US9541537B1 (en) | 2015-09-24 | 2017-01-10 | Frito-Lay North America, Inc. | Quantitative texture measurement apparatus and method |
US11243190B2 (en) | 2015-09-24 | 2022-02-08 | Frito-Lay North America, Inc. | Quantitative liquid texture measurement method |
US10107785B2 (en) | 2015-09-24 | 2018-10-23 | Frito-Lay North America, Inc. | Quantitative liquid texture measurement apparatus and method |
US10969316B2 (en) | 2015-09-24 | 2021-04-06 | Frito-Lay North America, Inc. | Quantitative in-situ texture measurement apparatus and method |
US10070661B2 (en) | 2015-09-24 | 2018-09-11 | Frito-Lay North America, Inc. | Feedback control of food texture system and method |
US10598648B2 (en) | 2015-09-24 | 2020-03-24 | Frito-Lay North America, Inc. | Quantitative texture measurement apparatus and method |
CN106370104B (zh) * | 2016-09-14 | 2019-01-18 | 华南理工大学 | 一种测量曲面led荧光粉涂层体积的方法 |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0671006B1 (de) * | 1992-11-26 | 1997-02-05 | Biolab Gmbh | Immunologischer schnelltest zur optischen bestimmung von progesteron in flüssigkeiten |
WO2003048771A2 (en) * | 2001-12-04 | 2003-06-12 | Lattec I/S | Device for analysing analyte compounds and use hereof |
Family Cites Families (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4778751A (en) * | 1986-05-12 | 1988-10-18 | Diagnostic Products Corporation | Method for measuring antigens or antibodies in biological fluids using ligand labeled antigens or ligand labeled antibodies |
US5156976A (en) * | 1991-06-07 | 1992-10-20 | Ciba Corning Diagnostics Corp. | Evanescent wave sensor shell and apparatus |
ATE377751T1 (de) * | 1995-05-12 | 2007-11-15 | Novartis Erfind Verwalt Gmbh | Verfahren zur parallelen bestimmung mehrerer analyten mittels evaneszent angeregter lumineszenz |
DE19628002C1 (de) * | 1996-07-11 | 1997-12-18 | Inst Chemo Biosensorik | Vorrichtung und Verfahren zur Durchführung von Fluoreszenzimmunotests |
US5922537A (en) * | 1996-11-08 | 1999-07-13 | N.o slashed.AB Immunoassay, Inc. | Nanoparticles biosensor |
US20030113939A1 (en) * | 1998-04-15 | 2003-06-19 | Bodenseewerk Perkin-Elmer Gmbh | Modified surface for carrying out or detecting affinity reactions |
US6300638B1 (en) * | 1998-11-12 | 2001-10-09 | Calspan Srl Corporation | Modular probe for total internal reflection fluorescence spectroscopy |
AU2002224831A1 (en) * | 2000-11-17 | 2002-05-27 | Zeptosens Ag | Kit and method for determining multiple analytes |
-
2005
- 2005-06-10 DE DE102005026839A patent/DE102005026839A1/de not_active Ceased
-
2006
- 2006-05-26 NZ NZ564074A patent/NZ564074A/en not_active IP Right Cessation
- 2006-05-26 WO PCT/EP2006/005078 patent/WO2006131225A1/de active Application Filing
- 2006-05-26 US US11/921,971 patent/US20100297671A1/en not_active Abandoned
- 2006-05-26 EP EP06743076A patent/EP1894007A1/de not_active Ceased
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0671006B1 (de) * | 1992-11-26 | 1997-02-05 | Biolab Gmbh | Immunologischer schnelltest zur optischen bestimmung von progesteron in flüssigkeiten |
WO2003048771A2 (en) * | 2001-12-04 | 2003-06-12 | Lattec I/S | Device for analysing analyte compounds and use hereof |
Non-Patent Citations (1)
Title |
---|
See also references of WO2006131225A1 * |
Also Published As
Publication number | Publication date |
---|---|
WO2006131225A1 (de) | 2006-12-14 |
DE102005026839A1 (de) | 2006-12-21 |
NZ564074A (en) | 2010-01-29 |
US20100297671A1 (en) | 2010-11-25 |
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