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WO1993003374A2 - Procede et dispositif d'analyse de reactions agglutinantes - Google Patents

Procede et dispositif d'analyse de reactions agglutinantes Download PDF

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Publication number
WO1993003374A2
WO1993003374A2 PCT/DE1992/000601 DE9200601W WO9303374A2 WO 1993003374 A2 WO1993003374 A2 WO 1993003374A2 DE 9200601 W DE9200601 W DE 9200601W WO 9303374 A2 WO9303374 A2 WO 9303374A2
Authority
WO
WIPO (PCT)
Prior art keywords
agglutination reactions
reactions according
analyzing
arrangement
analysis
Prior art date
Application number
PCT/DE1992/000601
Other languages
German (de)
English (en)
Other versions
WO1993003374A3 (fr
Inventor
Anton Horn
Barbara Horn
Heindrun Rhode
Original Assignee
Friedrich-Schiller-Universität Jena
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 Friedrich-Schiller-Universität Jena filed Critical Friedrich-Schiller-Universität Jena
Publication of WO1993003374A2 publication Critical patent/WO1993003374A2/fr
Publication of WO1993003374A3 publication Critical patent/WO1993003374A3/fr

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
    • B01L3/5025—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures for parallel transport of multiple samples
    • B01L3/50255—Multi-well filtration
    • 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
    • G01N33/536—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase
    • G01N33/537—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase with separation of immune complex from unbound antigen or antibody
    • G01N33/538—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase with separation of immune complex from unbound antigen or antibody by sorbent column, particles or resin strip, i.e. sorbent materials
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/26—Conditioning of the fluid carrier; Flow patterns
    • G01N30/38—Flow patterns
    • G01N2030/381—Flow patterns centrifugal chromatography
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
    • G01N2030/8809—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample
    • G01N2030/8813—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample biological materials
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/26—Conditioning of the fluid carrier; Flow patterns
    • G01N30/38—Flow patterns
    • G01N30/46—Flow patterns using more than one column
    • G01N30/466—Flow patterns using more than one column with separation columns in parallel
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/50—Conditioning of the sorbent material or stationary liquid
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/60—Construction of the column
    • G01N30/6052—Construction of the column body

Definitions

  • the invention relates to a method and an arrangement for analyzing agglutination reactions in medicine and biology.
  • Agglutination reactions are traditionally carried out on a large scale in blood group serology. Agglutination tests for a number of proteins and other ligands have become increasingly known. For these tests, antibodies to the substance to be determined are usually bound to specially prepared erythrocytes or defined particles, very often latex particles. There are a variety of possible variations when using such tests. The agglutination reaction is predominantly evaluated semi-quantitatively by visual observation or after dilution of at least one of the reactants in titer levels.
  • erythrocytes and other cells can be separated from low molecular weight accompanying substances in suitable media by means of molecular sieve gels. Furthermore, it is known that agglutinates from erythrocytes can only pass through certain agarose gels with difficulty. By visual inspection of such gel arrangements, which are designed as closed centrifuge tubes, agglutination reactions with erythrocytes can be assessed semi-quantitatively.
  • the invention has for its object a cost-effective, practical, widely applicable and automatable analysis system for
  • Reactants are introduced into a micro column that the agglutination reaction takes place in the micro column, that after the agglutination reaction there is a separation of
  • the reactants are placed in a micro column.
  • Agglutination reaction takes place in the microcolumn, preferably before the reactants pass through the filtering substance.
  • Particles are preferably separated by force-accelerated filtration.
  • the separation of particles by particle size can be done either by molecular sieve gels or by filtration, either through special filters or through filter aids. Gravity, centrifugal force, hydrostatic pressure or air pressure can be used to move the material to be separated. An electrical or magnetic field can also be used for separation.
  • the use of the method is expanded by the fact that at least one reaction partner is firmly connected to a marker substance.
  • marker substances can be radioactive isotopes, enzymes, luminescent markers, fluorescent markers or biospecific ligands with high affinity for substances that can be easily detected analytically. Marking with magnetic particles leads to a further possibility of expanding the use of the system.
  • the coupling of reactants to latex or other particles opens up the use of known particle agglutination techniques.
  • the measurement of the natural properties of participants for example the cloudiness of the solution of erythrocyte suspensions, ensures use in blood group serology and forms the basis for universal blood group automatons.
  • the analysis of the agglutination reaction can either be carried out directly by optical measurements of the properties of the labeled or unlabeled reactants, or after the interposition of an auxiliary step.
  • the layered arrangement of various chromatography materials in the micro column enables the pre-separation of various accompanying substances.
  • An extension of the entrance part of the micro column allows the reactants to be mixed.
  • the arrangement of the individual columns and collecting vessels in certain rows and matrix arrangements allows a high sample throughput and the automation of the process.
  • the fitable arrangement of the microcolumns to microtiter plates is particularly favorable, since commercially available microtiter plate readers can be used for many analytical questions.
  • the entire arrangement can be used in centrifuge rotors if the material is separated by centrifugal force.
  • the material separation can advantageously also be carried out by means of liquid pressure or air pressure.
  • the invention is illustrated by 9 exemplary embodiments.
  • FIG. 1 Plurality of microcolumns (microcolumn chamber), collecting vessels (microtiter plate) and pipettes (multichannel pipettes)
  • Fig. La shows the arrangement for analyzing agglutination reactions consisting of a microcolumn (1), which is composed of an input-side part (2) and an output-side part (3), separated by a filter (4).
  • the input part contains an extension (5) for introducing and mixing the reactants and a column part (6).
  • the column passage (7) is collected in a collecting vessel (8).
  • Fig. Lb shows the arrangement with a filter element (9) and the filling of the column with a chromatography material (10).
  • Fig. Lc shows the arrangement with a filter element and two different chromatography materials (11 and 12).
  • Example 4 Micro column for chromatography and filtration.
  • Fig. Id shows the arrangement with a filter (4) and a chromatography material (10).
  • Fig. 2 shows the plurality of the arrangement of the micro-columns (1) in micro-column chambers (13), the collecting vessels (8) in Microtiter plates (14) and the pipette tips in multi-channel pipettes (15).
  • Example 6 Blood group determination in the ABO system by separating the agglutinated particles by filtration
  • the agglutinates remain on the filter surface, the nonagglutinated erythrocytes sediment from the outlet part into the microtiter plate.
  • the microtiter plate is shaken on a horizontally rotating shaker and evaluated in a reader at 405 nm.
  • the non-agglutinated erythrocytes suspended in the microtiter plate have the following extinctions:
  • Each column part (6) of the columns of the micro column chamber (13) is filled with 30 ⁇ l of Sephadex G 200 soaked in 0.9% NaCl as the chromatography material (10) by filling 150 ⁇ l of a 20% gel suspension into the columns and at 500 U centrifuged for 5 min / min. Then 20 ⁇ l of an antiserum dilution are pipetted onto the gel surface. 5 ⁇ l of the 5% erythrocyte suspension are pipetted into this. All pipetting steps (gel suspension, antiserum dilution,
  • Erythrocyte suspension are carried out with multichannel pipettes (8, 12 or 96-fold) (15).
  • the column chambers are centrifuged at 250 rpm for 10 min over a microtiter plate and the column outlet of each column is thus collected in a hole in the microtiter plate, which contains 100 ⁇ l 0.9% NaCl per hole.
  • the erytrocyte agglutinates are on the gel or in the upper parts of the gel. The non-agglutinated erythrocytes sediment into the microtiter plate.
  • microtiter plate is then shaken by means of a shaker and then measured at 405 nm in a microtiter plate reader or an equivalent photometer.
  • the non-agglutinated erythrocytes suspended in the microtiter plate have the following extinctions: Reaction of A erythrocytes
  • Each column part (6) of the columns of the micro column chamber (13) is filled with 30 ⁇ l of Biogel 200 soaked in 0.9% NaCl, as described in application example 7. 20 ⁇ l of Coombs serum (anti-human globulin) are pipetted onto the gel surface. 5 ⁇ l of the unwashed, sensitized erythrocytes from whole blood are also pipetted onto the gel surface from a 5% suspension. The column chambers are incubated for 15-30 min at room temperature and then centrifuged and measured photometrically as in Application Example 7.
  • the erythrocytes agglutinated with Coombs serum are located as a sharp band on the gel surface or in the upper half of the gel.
  • the non-agglutinated erythrocytes sediment into the microtiter plate.
  • the non-agglutinated erythrocytes suspended in the microtiter plate have the following extinctions:
  • Application example 9 determination of antistreptolysin antibodies by means of latex agglutination and particle separation by chromatography.
  • Each column part (6) of the columns of the micro column chamber (13) is filled with 30 ⁇ l of poured Sephadex G200 superfine, as described in application example 7. 20 ⁇ l of the serum sample to be examined are pipetted onto the gel surface. For this purpose, 5 ⁇ l streptolysin-O-labeled latex particles (diluted 1: 2 in 0.9% NaCl, from rheumajet ASO, biokit) are pipetted. After incubation for 10 min at room temperature, the microcolumn chambers are centrifuged as in application example 6 over a microtiter plate. The agglutinated latex particles remain on the gel surface or in the upper parts of the gel, the non-agglutinated particles sediment into the microtiter plate. The microtiter plate is shaken and evaluated in a reader at 380 nm.
  • the non-agglutinated latex particles suspended in the microtiter plate have the following extinctions:

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  • Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Hematology (AREA)
  • Biomedical Technology (AREA)
  • General Health & Medical Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Molecular Biology (AREA)
  • Urology & Nephrology (AREA)
  • Food Science & Technology (AREA)
  • Microbiology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Cell Biology (AREA)
  • Biochemistry (AREA)
  • Biotechnology (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)

Abstract

Un procédé et un dispositif permettent d'analyser des réactions agglutinantes entre les partenaires réactionnels introduits dans une microcolonne. La réaction agglutinante se produit dans la microcolonne, de préférence avant que les partenaires réactionnels ne traversent la matrice en gel. Les particules agglutinées et non agglutinées sont séparées de préférence par filtration accélérée par application d'une force. L'éluat contenant des particules non agglutinées est soumis à une analyse quantitative après avoir quitté la microcolonne.
PCT/DE1992/000601 1991-07-26 1992-07-25 Procede et dispositif d'analyse de reactions agglutinantes WO1993003374A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE19914124778 DE4124778A1 (de) 1991-07-26 1991-07-26 Verfahren und anordnung zur analyse von agglutinationsreaktionen
DEP4124778.7 1991-07-26

Publications (2)

Publication Number Publication Date
WO1993003374A2 true WO1993003374A2 (fr) 1993-02-18
WO1993003374A3 WO1993003374A3 (fr) 1993-05-27

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ID=6437055

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/DE1992/000601 WO1993003374A2 (fr) 1991-07-26 1992-07-25 Procede et dispositif d'analyse de reactions agglutinantes

Country Status (3)

Country Link
AU (1) AU2374592A (fr)
DE (1) DE4124778A1 (fr)
WO (1) WO1993003374A2 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0678745A1 (fr) * 1994-04-22 1995-10-25 Scibiex (Sarl) Dispositif et procédé d'analyse immunologique
WO1997026536A1 (fr) * 1996-01-16 1997-07-24 Bernd Pevec Dispositif et procede pour le dosage et/ou l'elimination d'une substance dans un echantillon
US6471860B1 (en) * 1998-03-12 2002-10-29 Miltenyi Biotech Gmbh Magnetic micro separation column and method of using it

Families Citing this family (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2702050B1 (fr) * 1993-02-26 1995-05-24 Boy Inst Jacques Procédé de groupage sanguin faisant appel à des réactions immunoenzymatiques.
NL1006680C2 (nl) * 1997-03-04 1998-09-07 Alexander Adrianus Moen Werkwijze voor het detecteren van de aanwezigheid van een immunologisch reactief molecuul in een monster.
DE19734135A1 (de) 1997-08-07 1999-02-11 Boehringer Mannheim Gmbh System zum Zurverfügungstellen biologischer Materialien
DE60119079T2 (de) 2000-01-31 2006-11-30 Emory University System und verfahren für immunologische assays
US20050123456A1 (en) * 2001-12-04 2005-06-09 Lutz Eichacker Device and method for processing biological or chemical substances or substance mixtures thereof
DE102004033811A1 (de) * 2004-07-12 2006-02-02 Salama, Abdulgabar, Prof. Dr. Verfahren zum einfachen und schnellen Nachweis von Zellen und Biomolekülen mit Hilfe paramagnetischer Partikel
DE102006002258B4 (de) * 2006-01-17 2008-08-21 Siemens Ag Modul zur Aufbereitung einer biologischen Probe, Biochip-Satz und Verwendung des Moduls
DE102006055358A1 (de) * 2006-11-23 2008-05-29 Omx Gmbh Verfahren zum quantitativen Vergleich von zwei oder mehr Proteinen
SG11201402709WA (en) * 2011-12-06 2014-06-27 Univ Bruxelles Method and device for assaying an antigen present on erythrocytes or an antibody binding to an antigen present on erythrocytes
CN105652014A (zh) * 2015-12-31 2016-06-08 合肥天生物技术研究所 一种直接抗人球蛋白检测卡的制备方法

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4425438A (en) * 1981-03-13 1984-01-10 Bauman David S Assay method and device
US5073484A (en) * 1982-03-09 1991-12-17 Bio-Metric Systems, Inc. Quantitative analysis apparatus and method
WO1986007463A1 (fr) * 1985-06-03 1986-12-18 American National Red Cross Analyse par floculation d'une surface de microtitrage (msf) destine a deceler la presence d'anticorps ou d'antigenes
DE3717211A1 (de) * 1987-05-22 1988-12-01 Diagen Inst Molekularbio Vorrichtung und verfahren zur trennung und reinigung von molekuelen
WO1989000290A1 (fr) * 1987-07-02 1989-01-12 In Vitro Technologies, Inc. Dispositif capillaire permettant l'immuno-analyse d'analytes multiples
JPH087215B2 (ja) * 1987-08-24 1996-01-29 シュティフツング・フュア・ディアグノスティッシュ・フォルシュンク 抗原および／又は抗体の検出方法および検出用の試験キット
CA2055095C (fr) * 1990-11-09 2003-05-13 Johnna B. Hawk Essai et dispositif d'agglutination sur colonne

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0678745A1 (fr) * 1994-04-22 1995-10-25 Scibiex (Sarl) Dispositif et procédé d'analyse immunologique
FR2719122A1 (fr) * 1994-04-22 1995-10-27 Scibiex Sarl Dispositif et procédé d'analyse immunologique.
US5746975A (en) * 1994-04-22 1998-05-05 Scibiex (Sarl) Apparatus for immunological analysis
WO1997026536A1 (fr) * 1996-01-16 1997-07-24 Bernd Pevec Dispositif et procede pour le dosage et/ou l'elimination d'une substance dans un echantillon
US6471860B1 (en) * 1998-03-12 2002-10-29 Miltenyi Biotech Gmbh Magnetic micro separation column and method of using it

Also Published As

Publication number Publication date
WO1993003374A3 (fr) 1993-05-27
DE4124778A1 (de) 1993-01-28
AU2374592A (en) 1993-03-02

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