EP0302933A4 - Dispositif jetable d'immunoanalyse et de test biochimique adapte pour une utilisation sur site et en laboratoire. - Google Patents

Dispositif jetable d'immunoanalyse et de test biochimique adapte pour une utilisation sur site et en laboratoire.

Info

Publication number
EP0302933A4
EP0302933A4 EP19880902712 EP88902712A EP0302933A4 EP 0302933 A4 EP0302933 A4 EP 0302933A4 EP 19880902712 EP19880902712 EP 19880902712 EP 88902712 A EP88902712 A EP 88902712A EP 0302933 A4 EP0302933 A4 EP 0302933A4
Authority
EP
European Patent Office
Prior art keywords
membrane
template
exit port
enclosed region
exit
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.)
Withdrawn
Application number
EP19880902712
Other languages
German (de)
English (en)
Other versions
EP0302933A1 (fr
Inventor
Michael G Gabridge
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.)
BIONIQUE LABORATORIES Inc
BIONIQUE LAB Inc
Original Assignee
BIONIQUE LABORATORIES Inc
BIONIQUE LAB Inc
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 BIONIQUE LABORATORIES Inc, BIONIQUE LAB Inc filed Critical BIONIQUE LABORATORIES Inc
Publication of EP0302933A1 publication Critical patent/EP0302933A1/fr
Publication of EP0302933A4 publication Critical patent/EP0302933A4/fr
Withdrawn legal-status Critical Current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
    • 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
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D61/00—Processes of separation using semi-permeable membranes, e.g. dialysis, osmosis or ultrafiltration; Apparatus, accessories or auxiliary operations specially adapted therefor
    • B01D61/14—Ultrafiltration; Microfiltration
    • B01D61/18—Apparatus therefor

Definitions

  • the present invention relates to an apparatus for biochemical testing, such as spot tests, im unoassays, and similar reaction sequence which may involve a colored or chromogenic end point.
  • this invention relates to a multi-well device in which the lower surface of each well is a microporous membrane. Removal of reagents deposited in the wells is accomplished by applying a gentle vacuum across an enclosed region under the membrane with a syringe or equivalent device.
  • Microtiter wells are used in biochemical, clinical and biological laboratories for a multitude of functions including immunoassays such as the RadioImmunoAssay (RIA) and Enzyme Linked ImmunoSorbent Assay (ELISA) .
  • immunoassays such as the RadioImmunoAssay (RIA) and Enzyme Linked ImmunoSorbent Assay (ELISA) .
  • RIA RadioImmunoAssay
  • ELISA Enzyme Linked ImmunoSorbent Assay
  • a critical specific reagent is tagged with an enzyme, and addition of the appropriate enzyme substrate results in the development of a visually detected chromogenic product.
  • Such assays are widely used to detect and quantify specific antibodies in serum.
  • the antibody test sample is added to a microwell in a plastic plate previously coated with an antigen to which the antibody will attach in a specific fashion.
  • an antibody can be affixed to the wells to detect an antigen in test samples.
  • many devices incorporate a microp
  • One of the known designs is a two-piece ELISA filter device using multiple wells and membranes which are precoated with antigens or antibodies. Therefore, us® of this device is restricted to particular ligands.
  • the membranes are provided for regulation of fluid flow-through. Rather, the rate of fluid flow is governed by the hydrostatic pressure exerted on a drop of liquid deposited on the upper membrane surface as it rests on the top of the membrane. Reaction time is controlled by the resident time of the sample within the membrane pores.
  • FIG. 1 Another example of a known device used to assay liquid samples for the presence of a diagnostic reagent consists of a telescoping top and bottom member which define a liquid reservoir therebetween.
  • the top member has a multiplicity of test wells each having a bottom opening to the periphery to -which the top member has a coreactant immobilized on its internal and' external surfaces and permits passage of fluid through predetermined passageways.
  • a sorbent material is placed in the reservoir between the two members. After the liquid sample is placed in a fluid passageway on the top member r the top member is depressed to pass the sample through the membrane into the sorbent material.
  • the sorbent material is held in the base which consists of a surface layer preferably non-wetted by the liquids used and a bulk layer sufficiently thick to absorb all the liquids passing through the fluid passageways.
  • Still another apparatus is a reusable device consisting of a filter membrane clamped between a multi-well manifold and a vacuum chamber utilizing a sealing rubber gasket.
  • a minute hole in the well bottom allows capillary action to hold the ligand above the filter until a standard vacuum is applied to the vacuum nipple using a connecting hose.
  • the membrane may consist of an individual disk in each well, allowing removal of the individual disks, or a continuous sheet clamped between the upper element and its base.
  • Another device in use consists of an apparatus having a plurality of racks carrying cylindrical reaction vessels, each having a porous bottom connected to separate chambers.
  • the reaction vessels can be emptied simultaneously by suction from a combination vacuum pump and pressure-regulator.
  • the structures currently known are characterized by uneven and unreliable sealability between wells, resulting in well-to- well contamination. The lateral leakage is a serious failing because it obscures test results.
  • the complexity of previous, devices may require lengthy assembly time, frequently involving gaskets and clamps and/or screws to obtain a seal.
  • Filter misalignment and improper assembly lead to air leaks and contamination of wells by lateral migration and regulators as a means to withdraw fluids from reaction wells are expensive, bulky, require power to run and make fine control of pressure very difficult. These -characteristics make many of the known unsuitable for field use. Furthermore, the person performing the tests may be in contact with hazardous chemical solutions when the reservoirs are emptied and cleaned.
  • the present invention provides a disposable biochemical test apparatus for use in chromogenic filter-based assays and static assays.
  • the apparatus comprises an upper surface, a side wall and a bottom surface to form an enclosed region.
  • the upper surface comprises a planar template having top and lower surfaces and containing a plurality of spaced .apertures extending from the top surface through the template to the lower surface, each aperture being larger in the top surface and smaller in the lower surface, thereby providing conical sloping sides in the aperture.
  • a microporous membrane is placed in the enclosed region so as to underlie the template, the membrane having sufficient dimension to cover the lower apertures.
  • An exit port is provided to enable fluids to exit the enclosed region.
  • An exit ramp covering the bottom surface and slanting upward from- the exit port, is also provided.
  • the one-piece disposable test apparatus of the present invention is designed to provideaa plurality of discrete wells arranged in a horizontal ' array to permit a multitude of biochemical tests to be run simultaneously.
  • the device overcomes the disadvantages of similar devices previously known by its simplicity of design and ease of operation.
  • the device generally is comprised of a clear plastic block having a plurality of wells in its upper surface with the wells sloping inwardly. The sloping nature of the upper wells allows one to view the entire array of well bottoms and make visual or instrumental observations with ease.
  • the well bottoms are covered by a porous cellulose nitrate membrane, for example, and the lower surface of the enclosed region is ramped to an exit port.
  • the exit ramp commences near the lower apertures and slants -downward toward the exit port to enhance the migration of liquids to the exit port.
  • a vacuum is created in the enclosed region to move liquids in the wells " through the membrane toward the exit port of the apparatus. Flow through the wells is under operator control. All reactants and wash fluids are moved through the exit port to a reservoir and the entire assembly is disposed of without exposing the operator to any of the chemicals contained therein.
  • the invention is designed to be simple, inexpensive to manufacture and may be comprised of two or more molded plates sealed together resulting in a one-piece disposable unit. Without any necessity for a vacuum pump and line current, the unit is especially well adapted to office and field use.
  • Fig. 1 is a perspective view of one embodiment of the present invention
  • Fig. 2 is a cross-sectional view taken along line 2—2 of Fig. 1;
  • Fig. 3 is an end view of the embodiment of Fig. 1;
  • Fig. 4 is a perspective view of'another embodiment of the present invention;
  • Fig. 5 is a cross-sectional view taken along line 5—5 of Fig. 4;
  • Fig. 6 is a perspective view ⁇ of a futher embodiment of the present invention
  • Fig. 7 is a cross-sectional view taken along line 7—7 of Fig. 6.
  • FIG. 1 depicts a disposable biochemical test apparatus 10 having an upper surface 13 containing apertures or wells 12 permitting fluids to contact a membrane 14 and pass through the membrane 14 into an enclosed region provided by the side walls 15, the upper surface 13, the bottom surface 11 and the end plates 16 and 17.
  • the exit ramp 18 permits the liquid to migrate toward the exit port 19.
  • Fig. 2 shows the test apparatus of Fig. 1 in a cross- sectional view taken along line 2—2.
  • the upper surface 13 contains apertures 12 which are tapered conical apertures with the smaller aperture at the bottom contacting a microporous membrane 14.
  • the liquid passes through the membrane 14 and is deposited on the exit ramp 18.
  • the liquids thereby exit at the exit port 19 into a syringe or like ⁇ instrument which has provided suction and vacuum to draw the liquids through the membrane 14, the enclosed region 20, and exit port 19 in succession.
  • the bottom surface 11, the end plates 16 and 17, and the side walls 15 along with the top surface 13 provide the enclosed region 20.
  • the test apparatus and the syringe or like instrument containing the withdrawn liquid are disposed of after the results are recorded.
  • Fig. 3 is an end view through a clear plastic end plate 16.
  • the upper surface 13 contains the apertures or wells 12 which exit onto a microporous membrane 14.
  • the side walls 15, the bottom 11, the top surface 13 and the end plates as depicted by 16, provide the enclosed region 20 containing any liquids which may have migrated through the membrane within the interior of the structure.
  • Fig. 4 is a perspective view of a test apparatus 40 having an upper surfa-ce 43 in the shape of a frustum of a cone, in this case a circle, which has apertures or wells 42, a side wall 45, a bottom surface 41 and an exit port 49.
  • Fig. 5 is a cross-sectional view taken along line 5—5 of Fig. 4.
  • the lower portions of apertures 42 contact a membrane 46.
  • the upper surface 43 , the side wall 45, and the bottom surface 41 provide an enclosed region 44.
  • a funnel- shaped exit ramp 48 is provided to assist in migration of liquids from the membrane 46 toward the exit port 49.
  • Fig. 6 is a perspective view of a test apparatus 60 which contains within it an integral plunger 68 and a reservoir 67 and 72 which enables use without the necessity of an external syringe or like instrument.
  • the apparatus 60 has apertures 62 which are covered on the bottom by a membrane 63.
  • An exit ramp 65 lies below the membrane 63 and forms the lower portion of an enclosed region 64 such that liquids passing through the membrane 63 and entering the enclosed region 64 collect via gravity and drain through the lowermost port toward exit port 66, which is continuous with a reservoir 67.
  • Within the reservoir 67 there is a tight fitting moveable plunger 68.
  • a shaft guide 70 helps align the plunger-shaft 69 as it draws back the plunger 68.
  • the fixed shaft guide 70 could be threaded, such that the shaft 69 and the plunger 68 are withdrawn by a circular motion of the flange 71 instead of a direct outward pull on the flange 71.
  • a typical embodiment of the present invention is a test apparatus containing eight wells spaced evenly apart in the top surface of a test plate, which test plate is a rectangular block approximately 2.5 cm by 14 cm by 1.5 cm high.
  • the substantially rectangular block is made of a clear rigid inert plastic such as Lexan to facilitate the viewing of the fluid motion and transfer.
  • the ends of the test apparatus are plates which are trapezoidal in shape, having the long side of the trapezoid at the base for stability of the apparatus when it is placed on a surface for use.
  • the shape of the apparatus may vary and thus need not be rectangular.
  • a circular embodiment is shown in Figs. 4 and 5.
  • Other shapes such as a square or a triangle are satisfactory.
  • the microporous membrane may be constructed of any media capable of immobilizing a biochemical species including antigens, antibodies, cells, precipitates and the like.
  • the pores are smaller than the particles one wishes to retain on the upper surface and also small enough so as to prevent the migration of liquid through the membrane by gravity alone. The pores are large enough, however, so that liquid will drain through the membrane when a gentle vacuum is applied.
  • Suitable materials for the membrane include cellulose acetate, cellulose nitrate, mixed cellulose esters, nitrocellulose, nylon, polytetrafluoroethylene, polyethylene, and polypropylene. Other suitable materials will be apparent to those skilled in the art.
  • the membrane has pores averaging 0.45 microns in size.
  • the membranes can be affixed to the template containing the apertures or wells in any one of several ways. Bonding methods include chemical bonding, for example with a solvent or adhesive, thermal bonding, for example, ultrasonic welding, or the membrane may be molded in place. The precise method is inmaterial so long as lateral migration of liquid in the membrane is prohibited.
  • the apertures may be arranged in any desirable configuration so long as the apertures are spaced, preferably spaced at least 1 cm from center-to-center from each other.
  • the size of the aperture as measured across the upper aperture is preferably about 7.5 mm in diameter; however, the size may vary according to need. INDUSTRIAL APPLICABILITY
  • each well or aperture empties through the microporous membrane onto a sloped ramp which slopes toward an exit port to facilitate drainage.
  • a vacuum is applied to the exit port to allow the removal of the accumulated fluids.
  • a 5- or 10-milliliter disposable syringe connected by a 5- to 13-cm length of inert plastic tubing facilitates fluid removal and allows fine control of fluid flow-through, i.e., pressure, to be easily accomplished.
  • the flow-through mode of operation is useful in procedures involving contacts between mobile and immobile species wherein the latter is either covalently bound to the membrane or is unable to penetrate it and the former is suspended in the fluid in the well of the membrane.
  • the static mode of operation initially uses no vacuum and is useful in procedures requiring prolonged contact of two or more reactants in the fluid phase. Once the reaction is complete, vacuum is applied and filtration is achieved by utilizing the flow-through operation.
  • a test apparatus is connected through its exit port to a plastic disposable 5- or 10-milliliter syringe with a 7.5-cm length of tubing. Samples from 50- to 250-microliter aliquots are placed in each of the apertures or wells. The samples may be incubated for 5 to 10 minutes at 20 to 37 degrees C followed by removal through gentle vacuum created by pulling the syringe plunger outwardly.
  • the test antigen may be a bacterial suspension or a soluble antigen bound to inert carrier particles.
  • a blocking agent is placed in the apertures. It may consist, for example, of 1% casein or 5% serum.
  • a blocking agent is placed in the apertures. It may consist, for example, of 1% casein or 5% serum.
  • a primary specific antibody After the antibody is added the samples are treated with buffer rinse. * In each of the steps vacuum is applies.
  • a secondary antibody with a marker such as an enzyme is added and is ⁇ followed by a buffer rinse.
  • an enzyme substrate is added which also is followed by a buffer rinse.
  • the plunger of the syringe is pulled outwardly just enough to cause the liquid in the wells to be pulled through the membrane and deposited in the enclosed region or in the reservoir of the syringe. At this point the test is completed and scores for the presence or absence of chromogenic or colored end products may be recorded by observation.
  • the volume of the enclosed region should be selected so that the application of vacuum supplied by a 5- or 10-milliliter syringe is sufficient to draw the fluids from the wells through the membranes and into the enclosed region. •Generally, the capacity of the syringe is sufficient to pull the liquids into the enclosed region after each step in the process.
  • the reservoir will be of sufficient size to apply vacuum after each step in the process. Thus the flange is pulled outwardly a little at a time until the procedure is completed thereby drawing any liquids residing in the wells through the membranes into the enclosed region and on into the reservoir.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Engineering & Computer Science (AREA)
  • Water Supply & Treatment (AREA)
  • Analytical Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Hematology (AREA)
  • Clinical Laboratory Science (AREA)
  • Automatic Analysis And Handling Materials Therefor (AREA)
  • Devices For Use In Laboratory Experiments (AREA)

Abstract

Appareil de test biochimique jetable destiné à une utilisation dans des analyses par filtre et des analyses statiques. Une forme préférée de l'appareil est sensiblement rectangulaire, comprenant une surface supérieure (13), des parois (15) latérales trapézoïdales de support assurant la stabilité, une surface de fond (11), et des plaques d'extrémité (16, 17) à chaque extrémité. La surface supérieure (13) forme un gabarit plan contenant six ouvertures ou puits (12) cylindriques coniques disposés de manière linéaire et s'étendant à travers le gabarit. Disposée sous le gabarit se trouve une membrane (14) microporeuse fixée à la surface inférieure du gabarit. Une des plaques d'extrémité (17) dispose d'un orifice de sortie (19) qui sert d'écoulement aux fluides venant de la région fermée. La surface de fond (11) forme une rampe (18) de sortie inclinée vers le haut depuis l'orifice de sortie (19) jusqu'à l'autre plaque (16) d'extrémité. L'appareil se présente sous la forme d'un élément unitaire pouvant être jeté après utilisation. Pendant l'utilisation une seringue aspire du fluide sous l'effet d'un léger vide à travers l'orifice (19) de sortie.
EP19880902712 1987-02-24 1988-02-24 Dispositif jetable d'immunoanalyse et de test biochimique adapte pour une utilisation sur site et en laboratoire. Withdrawn EP0302933A4 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US1804387A 1987-02-24 1987-02-24
US18043 1987-02-24

Publications (2)

Publication Number Publication Date
EP0302933A1 EP0302933A1 (fr) 1989-02-15
EP0302933A4 true EP0302933A4 (fr) 1990-04-10

Family

ID=21785950

Family Applications (1)

Application Number Title Priority Date Filing Date
EP19880902712 Withdrawn EP0302933A4 (fr) 1987-02-24 1988-02-24 Dispositif jetable d'immunoanalyse et de test biochimique adapte pour une utilisation sur site et en laboratoire.

Country Status (2)

Country Link
EP (1) EP0302933A4 (fr)
WO (1) WO1988006723A1 (fr)

Families Citing this family (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5334189A (en) * 1991-06-03 1994-08-02 Wade Stephen E Device for controlled diffusion of a chemical substance
US5217619A (en) * 1992-03-06 1993-06-08 J. T. Baker Inc. Liquid-solid extraction apparatus and method of using same
CA2192262C (fr) * 1995-12-08 2011-03-15 Yoshihide Hayashizaki Methode pour purifier et transferer a des systemes de separation/detection des echantillons de sequencage de l'adn; plaques utilisees a cette fin
US6303389B1 (en) 1997-06-27 2001-10-16 Immunetics Rapid flow-through binding assay apparatus and method therefor
WO2001019502A2 (fr) * 1999-09-17 2001-03-22 Millipore Corporation Carte tamis de grande capacite
US6742659B2 (en) 2000-05-18 2004-06-01 Millipore Corporation Multiple well plate with adhesive bonded filter
KR100923482B1 (ko) 2001-05-31 2009-10-27 폴 코포레이션 유체처리용 웰
WO2013106081A2 (fr) 2011-06-20 2013-07-18 Emd Millipore Corporation Procédé et appareil pour filtrer un échantillon liquide
CN105424864B (zh) * 2015-12-30 2017-06-20 宁波美成生物科技有限公司 微量进样装置
WO2017120464A1 (fr) * 2016-01-08 2017-07-13 Siemens Healthcare Diagnostics Inc. Élément chauffant pour réseau de capteurs

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0131934A2 (fr) * 1983-07-15 1985-01-23 Baxter Diagnostics Inc. Cartouche pour essai
WO1986003589A1 (fr) * 1984-12-07 1986-06-19 Pharmacia Ab Procede, appareil et systeme pour effectuer une analyse d'affinite biospecifique a l'aide d'une colonne munie d'une partie temoin
GB2176601A (en) * 1985-06-10 1986-12-31 Bio Rad Laboratories Test plate assembly defining discrete regions on a microporous membrane with low boundary distortion

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3356462A (en) * 1966-08-09 1967-12-05 Cooke Engineering Company Disposable microtitration plate
US3554704A (en) * 1969-02-10 1971-01-12 Cordis Corp Biological assay cell
DE2526295B2 (de) * 1975-06-12 1978-11-23 Hoechst Ag, 6000 Frankfurt Verfahren zur Herstellung einer Nutsche
US4090850A (en) * 1976-11-01 1978-05-23 E. R. Squibb & Sons, Inc. Apparatus for use in radioimmunoassays
US4427415A (en) * 1979-01-05 1984-01-24 Cleveland Patrick H Manifold vacuum biochemical test method and device
US4526690A (en) * 1983-02-04 1985-07-02 Millipore Corporation Apparatus for nucleic acid quantification
US4493815A (en) * 1983-07-28 1985-01-15 Bio-Rad Laboratories, Inc. Supporting and filtering biochemical test plate assembly

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0131934A2 (fr) * 1983-07-15 1985-01-23 Baxter Diagnostics Inc. Cartouche pour essai
WO1986003589A1 (fr) * 1984-12-07 1986-06-19 Pharmacia Ab Procede, appareil et systeme pour effectuer une analyse d'affinite biospecifique a l'aide d'une colonne munie d'une partie temoin
GB2176601A (en) * 1985-06-10 1986-12-31 Bio Rad Laboratories Test plate assembly defining discrete regions on a microporous membrane with low boundary distortion

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of WO8806723A1 *

Also Published As

Publication number Publication date
WO1988006723A1 (fr) 1988-09-07
EP0302933A1 (fr) 1989-02-15

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