WO1992011939A1 - Dispositif d'essai in vitro - Google Patents

Dispositif d'essai in vitro Download PDF

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Publication number
WO1992011939A1
WO1992011939A1 PCT/US1992/000090 US9200090W WO9211939A1 WO 1992011939 A1 WO1992011939 A1 WO 1992011939A1 US 9200090 W US9200090 W US 9200090W WO 9211939 A1 WO9211939 A1 WO 9211939A1
Authority
WO
WIPO (PCT)
Prior art keywords
well
cap
column
wells
assay
Prior art date
Application number
PCT/US1992/000090
Other languages
English (en)
Inventor
Emanuel Calenoff
Original Assignee
Emanuel Calenoff
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 Emanuel Calenoff filed Critical Emanuel Calenoff
Publication of WO1992011939A1 publication Critical patent/WO1992011939A1/fr

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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
    • B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
    • B01L3/5085—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above for multiple samples, e.g. microtitration plates

Definitions

  • the present invention is directed to devices for performing in vitro testing.
  • microtiter wells have flat bottoms.
  • the discs When performing assays which use discs, which often are permeated with e.g., an antigen or other analyte, the discs frequentl adhere to the bottom of the well. This adherence prevents both effective washing of all sides of the di and full surface area exposure to incubating solutions thereby providing unexpected variations in the results the assay.
  • the materials from which microtite wells are made are usually degraded by organic solvent used in the detection of radioactively labeled analyte
  • the assay disc is transferred manually from the wel s to other containers for the detection of radioactivity in the assay sample.
  • prior art assays have often utilized microtiter dishes coated with a substance to which analyte binds.
  • the coating process is subject to variability often resulting in fluctuating amounts of the substance to which the analyte binds being coated onto the microtiter dish.
  • quality control is expensive for one producing standardized assays using microtiter dishes since an entire dish must be sacrificed during quality control testing.
  • the device of the present invention provides a multiple of joined wells, i.e., two or more, in which the bottom of the inside of each well is not flat but is concave, so as to avoid the tendency of assay discs to adhere to the well bottom.
  • the wells may have one or more vertical grooves that extend to the concave bottom of the well. These grooves provide an escape vent for air bubbles that often become trapped beneath assay discs thereby ensuring complete access of the disc to incubating fluids and thorough distribution of fluid around the discs in the wells. Complete access of the disc to surrounding fluid is important not only during assays but also when assay results are being determined, e.g., using scintillation counter fluids.
  • the present invention provides caps that one may place on the top of each well.
  • the caps have an opening which is large enough to allow one to wash assay discs and to administer reagents into each well with ease (and, optionally, automatically) while being small enough to prevent the disc itself from leaving the vial.
  • the top of the caps are sized so as to enable caps to fit on two or more adjacent wells.
  • the caps can be configured so as to fit on adjacent wells either individually or in a contiguous strip or sheet. Contiguous strips of caps have the physical advantage of stabilizing a row of wells and the commercial advantage of being less expensive to produce.
  • the caps may be configured so as to provide an air-tight seal, which is especially important for long-term storage of material within the well.
  • the outside diameter of the cap may be slightly larger than the inside diameter of the well resulting in an air-tight seal.
  • the present invention is directed to a device for performing in vitro assays, which device is comprised of multiple wells with each well joined to at least one other well, in which each well is an essentially cylindrical, hollow vertical column, said column having an upper end with an opening to said column and a lower end which is closed so as to provide an assay well reservoir, said lower closed end being concave inside of said column and essentially planar outside of said column.
  • the present invention further comprises a cap, at least a part of said cap having an outside diameter equal to the inside diameter of said cylinder; said cap further comprising an annular upper lip capable of preventing said cap from completely entering said cylinder.
  • the cap further comprises a central bore providing communication into and out of said cylinder.
  • FIG. 1 is an elevation showing a side view of multiple wells of the device of the present invention, in a linear array.
  • FIG. 2 is an elevation showing a longitudinal cross-sectional view of an assay well with a cap.
  • FIG. 3 is an elevation showing a longitudinal cross-sectional view of two joined assay wells.
  • FIG. 4 is an elevation showing a horizontal cross-sectional view on line 4 - 4 of FIG. 1 showing two joined assay wells.
  • the term "assay well" means a reservoir of any convenient size for performing a desired assay.
  • the type of assay performed in the device is not critical, since the device may be designed to accommodate many different assays. Examples of assays that can be performed in the device include ELISA, RAST, DNA hybridization, RNA hybridization, DNA blotting, RNA blotting, protein blotting.
  • a column is "essentially cylindrical” when its sides are generally parallel to one another; certain tolerances are permissible as the side wall arrangement is not critical. One may deviate from the "essentially cylindrical” configuration described herein and still be within the general rubric of "reservoir” as intended by the present invention.
  • “multiple wells” are at least two wells, may be 3 or more wells, are preferably at least 8 wells, and may be at least 96 wells.
  • the multiples of wells may be in any convenient array, for example, linear, or circular, or in the form of a plate, for example, a 96-well plate. Each well in the multiple wells is joined to at least one other well.
  • the device of the present invention provides wells in which the bottom of the inside of the well is not flat but is concave, so as to avoid the tendency of assay discs to adhere to the well bottom. This non- adherence provides for effective washing of all sides of the disc and equal exposure of both disc surfaces to incubating solutions/analytes thereby increasing the sensitivity and precision of the assay.
  • the outside dimensions of the well have a flat bottom so as to allow the wells to stand upright.
  • the present invention provides a cap that one may place on the top of the well.
  • the cap has an opening through it, which is large enough to allow one to wash assay discs with ease while being small enough to prevent the disc itself from leaving the vial.
  • the caps are sized so as to enable caps to fit on two or more adjacent wells.
  • the wells of the present invention are ideally non-adsorptive to prevent analytes to be assayed from adhering nonspecifically to the well.
  • the wells may be made out of non-adsorptive materials, for example, poorl adsorptive plastics such as polypropylene, polyethylene, etc.
  • the wells may be mad non-adsorptive by coating.
  • Non-adsorptive treatments an materials are well known in the art and include silicon, and bovine serum albumin (BSA) .
  • the wells of the present invention may be made of materials that are not degraded by the solvents used in the assay procedure, e.g., in scintillation counter fluids.
  • the wells may be made of material that allows one to separate the individual wells so that an assay may be placed in a desired array in a well holder or other suitable support means.
  • the wells may be used as storage containers or may be used as incubation containers during a particular assay run.
  • any convenient means may be employed to seal the wells.
  • caps can be designed so as to provide an essentially air tight seal by, for example, having the diameter of the cap sufficiently large so that it abuts completely the inside surface of the well at the point that the well contacts the cap.
  • the wells may be sealed from above with adhesive strips or sheets of material such as transparent acetate.
  • the wells may be pre ⁇ loaded with assay discs, wetting solution, and/or other reagents designed for use in a particular assay.
  • the wells and/or caps of the present invention may be color- coded to enable the laboratory technician or other individual using the device to color key samples or assays.
  • a device for in vitro assays contains multiples of a well 1, which may be aligned at juncture 30 in a linear array. Several linear arrays may be placed within a convenient holder.
  • the wells 1 may also be precast into a block in a format which may contain, for example, 96 wells.
  • FIG. 2 which is an enlarged detai of FIG. 1, one can see a longitdudinal cross-sectional view of the well 1.
  • well 1 ha essentially parallel vertical walls forming a cylinder, having an outer wall 13 and an inner wall 12. One end o the cylinder is open and the other end is closed.
  • the closed end has a concave bottom 10 on the interior of th well 1 and optionally has a planar base 11 on the exterior of the well 1. Because assay discs typically have a size in the range of 5 to 6 mm, the bottom 10 diameter at its widest point will be about 0.2 mm larger than the size of the assay disc.
  • the concave bottom 10 prevents assay discs from adhering to the bottom of the well reservoir.
  • the planar base 11 permits one to stand a well upright.
  • the well 1 may be fitted with a cap 2.
  • the cap 2 has essentially cylindrical parallel sides forming an essentially cylindrical column with an outer wall 26 having a diameter dimensioned so a to fit within the inner wall 12 of well 1.
  • the cap 2 ha at one end of the column an annular lip 20 which has a diameter greater than the inner wall 12 of well 1 thereb preventing the cap 2 from completely entering the well 1
  • the lip 20 does not extend beyond the outer wall 13 of the well 1 so as to permit adjacent wells, as shown in FIG. 1 and FIG. 3, to each have a cap 2.
  • the cap is slightly tapered beneath the annular lip 20 so as to provide a void 22 between the cap 2 and the internal wal 12 of the well 1 at the cap's upper end and fit snugly against the internal wall 12 of well 1 at the cap's lowe end 23.
  • the cap 2 also may have a central bore 21 which, when the cap 2 is placed into well 1, permits communication between the outside of well 1 and the inside of well 1.
  • the bore 21 may be preferably about 0.2 mm smaller in diameter than an assay disc present in well 1 thereby preventing the assay disc in the well 1 from passing through the bore 21 and out of the well 1 during assay wash steps.
  • the standard assay disc has a size in the range of between about 5 mm and about 6 mm.
  • the bore 21 diameter in the cap 2 will preferably be a minimum size of about 4 mm.
  • the upper end 24 of the bore 21 can be tapered at an angle that is about 23° relative to the inside wall 25 of the lower end of the bore 21.
  • the inner wall 12 of well 1 may have a vertical groove 14. It is preferable that the bottom of the groove 14 be slightly above and sloping toward the bottom and center of the concave bottom 10 so that fluids may fully drain from the groove 14.
  • One or more grooves 14 may be present in well 1.
  • the primary function of the vertical groove 14 is to permit air bubbles to escape from beneath discs that may be placed in the well l.
  • the groove 14 provides an escape vent for air bubbles thereby ensuring complete access of the disc to incubating fluids and thorough distribution of fluid around the discs in the well l.
  • the well 1 is attached to an adjacent well 1 at a juncture 30.
  • the wells 1 may be constructed of a material, such as polystyrene, that allows the user of the wells to snap off one or more wells at juncture 30 thereby enabling one to rearrange the wells and thereby vary the array.
  • Wells can be individual or in sequence as part of a strip of wells each well containing a paper disc with different covalently coupled allergen from the next.
  • Step No. 8 twice. After third soaking step, wash discs twice and aspirate. 10. Pipette 50 ⁇ l of 125 I-labeled, affinity- purified anti-IgE through the cap aperture into each well. (Anti-IgE is diluted with solution containing 50 mM PBS + 5% Horse SErum + 0.1% NaN3.)
  • the above protocol may be automated to increas the efficiency further.
  • the present invention vastly increases the efficiency of performing the RAST test by decreasing the time for setting up the assay and washing the discs, and pipetting assay reagents into the wells.
  • scintillation counting of the assay ' results can be done in the same container in which the assay is performed.
  • This increased efficiency compares favorably to the commercially available ELISA assays which, unlike the RAST test, often have the disadvantage of being unable to low levels of specific IgE (which fall in modified RAST class I sensitivities) .
  • this device allows a saving in the amount of reagent required to wash assay discs.

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Hematology (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

Dispositif pour analyses in vitro, qui comprend des réservoirs d'analyse multiples, chaque réservoir (1) se composant essentiellement d'une colonne cylindrique creuse et verticale, ladite colonne comprenant une extrémité supérieure pourvue d'une ouverture, et une extrémité inférieure fermée de façon à produire un réservoir d'analyse, ladite extrémité inférieure fermée étant de forme concave (10) à l'intérieur de ladite colonne et essentiellement plane (11) à l'extérieur de ladite colonne. En outre, les réservoirs peuvent comprendre une ou plusieurs rainures verticales (14) qui s'étendent jusqu'au fond concave (10) du réservoir. La présente invention comprend aussi des capuchons (2) que l'on peut placer au-dessus de chaque réservoir. Les capuchons (2) sont pourvus d'un passage (21) s'ouvrant sur l'extérieur et suffisamment grande pour permettre à des solutions d'être aisément ajoutées, cependant le passage (21) est suffisamment petit pour empêcher que le disque lui-même ne s'échappe du réservoir. Le dispositif permet d'effectuer des analyses in vitro de façon plus efficace et plus économique.
PCT/US1992/000090 1991-01-11 1992-01-10 Dispositif d'essai in vitro WO1992011939A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US64175591A 1991-01-11 1991-01-11
US641,755 1991-01-11

Publications (1)

Publication Number Publication Date
WO1992011939A1 true WO1992011939A1 (fr) 1992-07-23

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Country Status (2)

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AU (1) AU1236392A (fr)
WO (1) WO1992011939A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2322121A (en) * 1997-02-13 1998-08-19 Porvair Plc Multi-well plate closure
WO1999020394A2 (fr) * 1997-10-17 1999-04-29 Texperts, Inc. Ensemble microplaque a confinement

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4111754A (en) * 1976-11-29 1978-09-05 Hydow Park Immunological testing devices and methods
US4528268A (en) * 1981-12-31 1985-07-09 H. W. Andersen Products Inc. Apparatus and method for testing the sufficiency of sterilization
US4537311A (en) * 1983-06-27 1985-08-27 Russell G. Sharp Seamless suppository dispenser
US4956150A (en) * 1985-11-27 1990-09-11 Alerchek Disposable microtiter stick
US4956298A (en) * 1988-01-13 1990-09-11 Stephan Diekmann Separation or reaction column unit
US5017341A (en) * 1988-04-19 1991-05-21 Olympus Optical Co., Ltd. Agglutination analyzing vessel

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4111754A (en) * 1976-11-29 1978-09-05 Hydow Park Immunological testing devices and methods
US4528268A (en) * 1981-12-31 1985-07-09 H. W. Andersen Products Inc. Apparatus and method for testing the sufficiency of sterilization
US4537311A (en) * 1983-06-27 1985-08-27 Russell G. Sharp Seamless suppository dispenser
US4956150A (en) * 1985-11-27 1990-09-11 Alerchek Disposable microtiter stick
US4956298A (en) * 1988-01-13 1990-09-11 Stephan Diekmann Separation or reaction column unit
US5017341A (en) * 1988-04-19 1991-05-21 Olympus Optical Co., Ltd. Agglutination analyzing vessel

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2322121A (en) * 1997-02-13 1998-08-19 Porvair Plc Multi-well plate closure
WO1999020394A2 (fr) * 1997-10-17 1999-04-29 Texperts, Inc. Ensemble microplaque a confinement
WO1999020394A3 (fr) * 1997-10-17 1999-07-29 Texperts Inc Ensemble microplaque a confinement

Also Published As

Publication number Publication date
AU1236392A (en) 1992-08-17

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