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WO1993013856A1 - Dispositif et procede de dosage - Google Patents

Dispositif et procede de dosage Download PDF

Info

Publication number
WO1993013856A1
WO1993013856A1 PCT/GB1993/000097 GB9300097W WO9313856A1 WO 1993013856 A1 WO1993013856 A1 WO 1993013856A1 GB 9300097 W GB9300097 W GB 9300097W WO 9313856 A1 WO9313856 A1 WO 9313856A1
Authority
WO
WIPO (PCT)
Prior art keywords
assay
conjugate
species
zone
assayed
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
Application number
PCT/GB1993/000097
Other languages
English (en)
Inventor
Adrian Parton
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Genera Technologies Ltd
Scientific Generics Ltd
Original Assignee
Genera Technologies Ltd
Scientific Generics Ltd
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 Genera Technologies Ltd, Scientific Generics Ltd filed Critical Genera Technologies Ltd
Priority to EP93902397A priority Critical patent/EP0621803A1/fr
Publication of WO1993013856A1 publication Critical patent/WO1993013856A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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
    • 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

Definitions

  • the present invention relates to assay apparatus and a method of assay and has particular but not exclusive relevance to immuno-diagnostic assays.
  • test kits currently available do not require many steps, they tend to be expensive as the technology needed to achieve these requirements is inherently expensive. There is therefore a need for a simpler device reducing manufacturing costs whilst maintaining simplicity of operation.
  • the present invention provides assay apparatus comprising a first assay zone on a first solid support component for capturing a species to be assayed thereon and a second assay zone on a second solid support component having an assay reagent thereon, means mounting said first and second solid support components together for relative slidingmovement such that said zones are slidable with respect to one another from a first position in which said zones are separated from one another to a position in which said zones are brought into mutual contact by sliding motion.
  • the first and second support components may be different parts of a single member or may be separate members mounted together.
  • a single strip may be folded over to bring to end portions together and means may be provided to hold the end portions face to face whilst allowing sliding movement thereof with respect to one another.
  • the first and second assay zones are provided on respective ones of a pair of plastics plate members constituting said first and second solid support components.
  • one said plate member or one end region of such a strip has a pair of oppositely directed channels thereon in which edge regions of the other said plate member are slidingly received to mount said plate members or said strip regions in face to face sliding relationship.
  • the species to be assayed may itself be such as to produce or catalyse an observable reaction with the assay- reagent in the second assay zone. Otherwise, an assay reagent may be present unbound in said first reagent support zone which is a species capable of binding to the species to be assayed having conjugated thereto a species capable of mediating an observable reaction in the presence of said assay reagent of the second assay zone.
  • the observable reaction is preferably a colour forming reaction or a reaction giving rise to a species which is observable by fluorescence under UV light.
  • one or both of the support components is preferably transparent at least in its respective assay zone.
  • the unbound assay reagent present in said first assay zone may be an antibody conjugate capable of binding to a species to be assayed.
  • antibody in this context herein includes antibody fragments having selective affinity for antibody binding sites.
  • It may be a conjugate between a nucleic acid or nucleic acid analogue for binding to a nucleic acid as the species to be assayed and a linker moiety such as biotin capable of binding to a conjugate between an enzyme for mediating such a reaction and a further linking moiety, e.g. avidin or between said nucleic acid component and said enzyme component directly.
  • a linker moiety such as biotin capable of binding to a conjugate between an enzyme for mediating such a reaction and a further linking moiety, e.g. avidin or between said nucleic acid component and said enzyme component directly.
  • the assay reagent in said second assay zone may be a substrate for a colour forming or other observable reaction mediated by said conjugate.
  • the species to be assayed is to be captured on to the first assay zone so that its attachment is resistant to washing procedures during the assay. In some cases this may be achieved by direct physical capture or absorption. For instance, some bacteria or proteins will bind directly onto a roughened plastics strip or mesh.
  • the first assay zone may have bound thereon an assay reagent which is capable of binding the species to be assayed.
  • This bound reagent may be an antibody, which in this context includes antibody molecules and also fragments thereof having selective binding affinity for antibody binding sites, a nucleic acid or a nucleic acid analogue, depending upon the nature of the species to be assayed.
  • the bound an /or the unbound assay reagent in the first assay zone is an antibody or antibody conjugate the antibody or antibody fragment is preferably a monoclonal antibody or fragment thereof.
  • all of the reagents are present in dry form, the solvent (usually water) needed for the assay, reaction being derived from the addition of the sample.
  • the solvent usually water
  • This pad may for instance be of open cell foam or of absorbent paper such as filter paper or similar materials.
  • the arrangement may be such that the pad is compressed when the second solid support component is slid into position over the first assay zone.
  • the invention includes a method of assay comprising capturing a species to be assayed on a first assay zone provided on a solid support component, and (a) sliding thereover a second support component bearing a substrate for an observable reaction mediated by said species to be assayed so as to contact said substrate with said bound species to produce said observable reaction or (b) binding thereto a conjugate between an enzyme and a species capable of binding to the species to be assayed, washing to remove unbound conjugate and sliding thereover a second solid support component bearing a substrate for an observable reaction mediated by said conjugate so as to contact said substrate with said bound conjugate to produce said observable reaction.
  • Figure 1 shows in perspective view an example of apparatus according to the invention in use in three stages of an assay procedure
  • Figure 2 shows stages, including sample taking, involved in a specific form of assay procedure described in Example 1 below using the same apparatus according to the invention.
  • apparatus comprises a first solid support component 10 in the form of a plastics plate of about the size of a credit card having a pair of upstanding walls 12 bordering a central strip portion of lesser thickness 14. • The faces of the walls 12 bordering the strip 14 are undercut as shown at 16.
  • a second solid support component 18 comprises a plastics strip dimensioned to fit snugly between the walls 12 and having chamfered edges 20 which locate in the undercut shown at 16 to mount the support component 18 for sliding movement in the channel formed between the walls 12 on the support component 10.
  • a first assay zone 22 is provided on the first solid support component 10 and a second assay zone 24 is provided on the second solid support component 18. At least the region of the said assay zone 24 on the second support component 18 is made transparent.
  • a foam pad 26 is present in the first assay zone 22 securely adhered in place on the first support component 10.
  • a substrate for an enzyme mediated colour forming reaction is present in dry form in the second assay zone 24.
  • the apparatus illustrated in Figure 1 further comprises unbound on the foam pad 26 an enzyme-antibody conjugate, the enzyme being suitable to catalyse a colour forming reaction with the substrate present on the second assay zone 24.
  • the bound antibody and the antibody component of the unbound antibody conjugate may be the same or may be different. Both are preferably monoclonal anti ⁇ bodies although polyclonal reagents are still suitable.
  • a sample such as a liquid containing micro ⁇ organism cells is applied to the foam pad.
  • the cells are bound by the bound antibody and a "sandwich" is formed by binding to the bound cells of the antibody conjugate.
  • the apparatus may then be washed under running water, which may be tap water, to remove excess antibody conjugate from the sample area and the second support component 18 is then slid over the first support component 10 to bring the first and second assay zones into position one over the other.
  • the foam filter pad is thereby compressed against the underside of the second support component. Excess liquid is squeezed out from the foam pad by the leading edge of the second support component passing over it and a good contact is ensured between the foam pad 26 and the second assay zone 24 which contains in dried form a substrate for the antibody conjugate. This is resuspended by the residual liquid on the foam pad and the colour forming reaction takes place and is observable through the transparency of the second support component.
  • the following examples illustrate the use of the apparatus shown in Figure 1.
  • EXAMPLE 1 Assay for E.coli by Detection of ⁇ -Glucuronidase
  • ⁇ - glucuronidase is an enzyme expressed by a limited number of micro-organisms which is accepted in the water industry as being a 95% positive test for E. coli.
  • a sample of water is taken up into a syringe 27 and is discharged via a micro-organism trapping 0.2 ⁇ m filter 28.
  • the micro-organisms initially present in the liquid sample are now held in concentrated form in the liquid in the dead space of the syringe/filter assembly.
  • a defined amount of a suitable micro-organism growth medium is drawn up into the filter.
  • the syringe/filter assembly is now placed with the syringe vertical and the lower face of the housing of the filter on a heating block by which the filter is warmed to an incubating temperature. After a predetermined period of incubation to multiply the bacteria, the temperature of the heating block may be raised to a temperature suitable to produce lysis of the bacteria for a period of about one minute.
  • Apparatus according to Figure 1 is employed having a polyclonal antibody to ⁇ -glucuronidase bound to the foam pad 26. In this case no antibody conjugate is present on the foam pad as ⁇ -glucuronidase itself catalyses a colour forming reaction.
  • the liquid content of the syringe/filter assembly may then be discharged on to the first assay zone 22 of apparatus as shown in Figure 1 and may be left at room temperature for a period sufficient to enable binding of the ⁇ -glucuronidase from the micro-organism lysate by the bound antibody on the first assay zone, e.g. for abou-t ten minutes.
  • the assay apparatus is optionally then washed and the second support component 18 is slid into position as shown in step 3 in Figure 1. The colour forming reaction is allowed to take place and is observed.
  • the growth medium may contain a substrate suitable to induce expression of the ⁇ -glucuronidase by the micro-organisms to produce elevated levels of enzyme in the cultured cells, e.g. an R- ⁇ -D-glucuronide.
  • the colour forming reaction may be replaced by a reaction giving rise to a UV-fluorescent compound by appropriate selection of the enzyme substrate on the second support component 18.
  • Example 1 To enhance sensitivity the apparatus employed in Example 1 is modified by the inclusion of a conjugate between an anti ⁇ -glucuronidase antibody and a colour forming reaction catalysing enzyme in dried form unbound in said foam pad 26.
  • the procedure followed is as in Example 1 up to the end of the lysis step.
  • the liquid content of the syringe/filter assembly is discharged on to the foam pad a sandwich is formed in which the conjugate and the bound antibody both bind to the ⁇ -glucuronidase.
  • the foam pad is washed to remove unbound conjugate and the second support component is slid into position.
  • the assay reagent thereon is selected to take part in a colour forming reaction in the presence of the bound conjugate.
  • Example 2 The procedure of Example is modified in that the bound antibody on the foam pad 26 is omitted and the foam material is selected such that it has a non-specific affinity for proteins including ⁇ -glucuronidase.
  • the lysate as applied to the foam pad and proteins therein are bound to the foam.
  • Bound ⁇ -glucuronidase is further bound by the free antibody conjugate.
  • the presence of bound conjugate is then detected by washing and developing as in Example 2.
  • the antibody conjugate is also omitted and the trapped ⁇ - glucuronidase acts directly to catalyse the colour forming reaction as in Example 1.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Immunology (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)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

Dispositif de dosage comprenant une première zone de dosage constituée par une pastille en mousse (26) située sur un premier élément de support solide (10) constitué par une carte en plastique possédant un canal (14) sur une de ses faces bordée par des parois comportant une gorge (12), de façon à recevoir une glissière en plastique (18) en tant que deuxième élément de support solide possédant une fenêtre (24) en tant que deuxième zone de dosage. Les éléments de support s'adaptent l'un à l'autre, afin d'effectuer un déplacement coulissant relatif, de façon que lesdites zones coulissent l'une par rapport à l'autre depuis une première position dans laquelle elles sont séparées l'une de l'autre vers une position dans laquelle elles viennent en contact réciproque au moyen d'un déplacement coulissant. Un anticorps peut être fixé à la pastille en mousse. Un conjugué constitué par un anticorps et par un enzyme peut se trouver non fixé sur ladite pastille. Un réactif de dosage se trouve sous la fenêtre (24), de façon à servir dans une réaction observable avec le conjugué ou avec une espèce à analyser.
PCT/GB1993/000097 1992-01-18 1993-01-15 Dispositif et procede de dosage Ceased WO1993013856A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP93902397A EP0621803A1 (fr) 1992-01-18 1993-01-15 Dispositif et procede d'essai

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB929201089A GB9201089D0 (en) 1992-01-18 1992-01-18 A diagnostic article
GB9201089.1 1992-01-18

Publications (1)

Publication Number Publication Date
WO1993013856A1 true WO1993013856A1 (fr) 1993-07-22

Family

ID=10708832

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/GB1993/000097 Ceased WO1993013856A1 (fr) 1992-01-18 1993-01-15 Dispositif et procede de dosage

Country Status (5)

Country Link
EP (1) EP0621803A1 (fr)
AU (1) AU3359193A (fr)
CA (1) CA2127980A1 (fr)
GB (1) GB9201089D0 (fr)
WO (1) WO1993013856A1 (fr)

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5708008A (en) * 1995-03-20 1998-01-13 Eli Lilly And Company 5-Substituted-3-(1,2,3,6-tetrahydropyridin-4-yl)-and 3-(piperidin-4-yl)-1H-indoles: new 5-HT1F agonists
US5744096A (en) * 1997-02-21 1998-04-28 Cholestech Corporation Automated immunoassay cassette
WO2002064252A1 (fr) * 2001-02-15 2002-08-22 Technische Universiteit Delft Plaque de reaction a couvercle glissant et procede d'utilisation correspondant
WO2003041863A3 (fr) * 2001-11-16 2003-12-04 Univ Delft Tech Procede de remplissage d'un puits dans un substrat
WO2004047979A1 (fr) * 2002-11-21 2004-06-10 Corning Incorporated Dispositifs pour reactions biologiques et chimiques, et leurs procedes de production
JP2008544292A (ja) * 2005-06-28 2008-12-04 キンヌネン，パーヴォ 液間界面を形成するための、特にその表面張力を測定するための方法及び装置
US7795038B2 (en) 2002-04-09 2010-09-14 Cholestech Corporation High-density lipoprotein assay device and method
WO2010109445A1 (fr) 2009-03-23 2010-09-30 Ramot At Tel-Aviv University Ltd. Dispositif et procede de dosage
US7824879B2 (en) 2007-01-09 2010-11-02 Cholestech Corporation Device and method for measuring LDL-associated cholesterol
EP2700449A1 (fr) * 2012-08-22 2014-02-26 Morinaga & Co., Ltd. Dispositif immunochromatographique

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5449794A (en) * 1991-02-15 1995-09-12 Jasmine Fockerman Benzopyran phenol derivatives for use as antibacterial agents

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR1549783A (fr) * 1966-12-22 1968-12-13
EP0217403A2 (fr) * 1985-10-04 1987-04-08 Abbott Laboratories Dispositif analytique en phase solide et procédé pour son utilisation
FR2609334A1 (fr) * 1987-01-05 1988-07-08 Dole Assoc Inc Dispositif d'immuno-essai ou de diagnostic et son procede de fabrication
EP0299359A2 (fr) * 1987-07-16 1989-01-18 Abbott Laboratories Système pour délivrer des réactifs utilisés dans des dispositifs analytiques de phase solide
US4859604A (en) * 1987-08-27 1989-08-22 Ampor, Inc. Composition for stabilization of diagnostic reagents
EP0353025A2 (fr) * 1988-07-27 1990-01-31 Eastman Kodak Company Dispositif de test glissant pour essais
EP0369836A1 (fr) * 1988-10-11 1990-05-23 Institut Textile De France Dispositif à usage unique pour tests biologiques
EP0490447A1 (fr) * 1990-12-10 1992-06-17 Financieringsmaatschappij Sparrendaal I B.V. Dispositif jetable d'analyse de liquide

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR1549783A (fr) * 1966-12-22 1968-12-13
EP0217403A2 (fr) * 1985-10-04 1987-04-08 Abbott Laboratories Dispositif analytique en phase solide et procédé pour son utilisation
FR2609334A1 (fr) * 1987-01-05 1988-07-08 Dole Assoc Inc Dispositif d'immuno-essai ou de diagnostic et son procede de fabrication
EP0299359A2 (fr) * 1987-07-16 1989-01-18 Abbott Laboratories Système pour délivrer des réactifs utilisés dans des dispositifs analytiques de phase solide
US4859604A (en) * 1987-08-27 1989-08-22 Ampor, Inc. Composition for stabilization of diagnostic reagents
EP0353025A2 (fr) * 1988-07-27 1990-01-31 Eastman Kodak Company Dispositif de test glissant pour essais
EP0369836A1 (fr) * 1988-10-11 1990-05-23 Institut Textile De France Dispositif à usage unique pour tests biologiques
EP0490447A1 (fr) * 1990-12-10 1992-06-17 Financieringsmaatschappij Sparrendaal I B.V. Dispositif jetable d'analyse de liquide

Cited By (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5708008A (en) * 1995-03-20 1998-01-13 Eli Lilly And Company 5-Substituted-3-(1,2,3,6-tetrahydropyridin-4-yl)-and 3-(piperidin-4-yl)-1H-indoles: new 5-HT1F agonists
US5744096A (en) * 1997-02-21 1998-04-28 Cholestech Corporation Automated immunoassay cassette
WO2002064252A1 (fr) * 2001-02-15 2002-08-22 Technische Universiteit Delft Plaque de reaction a couvercle glissant et procede d'utilisation correspondant
WO2003041863A3 (fr) * 2001-11-16 2003-12-04 Univ Delft Tech Procede de remplissage d'un puits dans un substrat
CN1329125C (zh) * 2001-11-16 2007-08-01 代尔夫特工业大学 充填衬底中凹坑的方法
US7795038B2 (en) 2002-04-09 2010-09-14 Cholestech Corporation High-density lipoprotein assay device and method
WO2004047979A1 (fr) * 2002-11-21 2004-06-10 Corning Incorporated Dispositifs pour reactions biologiques et chimiques, et leurs procedes de production
JP2008544292A (ja) * 2005-06-28 2008-12-04 キンヌネン，パーヴォ 液間界面を形成するための、特にその表面張力を測定するための方法及び装置
EP1896182A4 (fr) * 2005-06-28 2010-09-08 Paavo Kinnunen Procédé et dispositif de formation d une interface liquide/liquide, en particulier pour une mesure de tension superficielle
US7824879B2 (en) 2007-01-09 2010-11-02 Cholestech Corporation Device and method for measuring LDL-associated cholesterol
WO2010109445A1 (fr) 2009-03-23 2010-09-30 Ramot At Tel-Aviv University Ltd. Dispositif et procede de dosage
EP2700449A1 (fr) * 2012-08-22 2014-02-26 Morinaga & Co., Ltd. Dispositif immunochromatographique
JP2014041061A (ja) * 2012-08-22 2014-03-06 Morinaga & Co Ltd イムノクロマトグラフィー用デバイス
CN103630680A (zh) * 2012-08-22 2014-03-12 森永制果株式会社 免疫层析用装置

Also Published As

Publication number Publication date
CA2127980A1 (fr) 1993-07-22
AU3359193A (en) 1993-08-03
GB9201089D0 (en) 1992-03-11
EP0621803A1 (fr) 1994-11-02

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