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WO1996019739A2 - Dispositif de dosage - Google Patents

Dispositif de dosage Download PDF

Info

Publication number
WO1996019739A2
WO1996019739A2 PCT/EP1995/005132 EP9505132W WO9619739A2 WO 1996019739 A2 WO1996019739 A2 WO 1996019739A2 EP 9505132 W EP9505132 W EP 9505132W WO 9619739 A2 WO9619739 A2 WO 9619739A2
Authority
WO
WIPO (PCT)
Prior art keywords
support
solid
assay
support according
elements
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/EP1995/005132
Other languages
English (en)
Other versions
WO1996019739A3 (fr
Inventor
Johan Haggblad
Marie ÖSTERLUND
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.)
Karo Healthcare AB
Original Assignee
Karo Bio AB
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 Karo Bio AB filed Critical Karo Bio AB
Priority to AU47135/96A priority Critical patent/AU4713596A/en
Publication of WO1996019739A2 publication Critical patent/WO1996019739A2/fr
Publication of WO1996019739A3 publication Critical patent/WO1996019739A3/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
    • B01L3/5023—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures with a sample being transported to, and subsequently stored in an absorbent for analysis
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01T—MEASUREMENT OF NUCLEAR OR X-RADIATION
    • G01T1/00—Measuring X-radiation, gamma radiation, corpuscular radiation, or cosmic radiation
    • G01T1/16—Measuring radiation intensity
    • G01T1/20—Measuring radiation intensity with scintillation detectors
    • G01T1/203—Measuring radiation intensity with scintillation detectors the detector being made of plastics
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01T—MEASUREMENT OF NUCLEAR OR X-RADIATION
    • G01T1/00—Measuring X-radiation, gamma radiation, corpuscular radiation, or cosmic radiation
    • G01T1/29—Measurement performed on radiation beams, e.g. position or section of the beam; Measurement of spatial distribution of radiation
    • G01T1/2914—Measurement of spatial distribution of radiation
    • G01T1/2921—Static instruments for imaging the distribution of radioactivity in one or two dimensions; Radio-isotope cameras
    • G01T1/2942—Static instruments for imaging the distribution of radioactivity in one or two dimensions; Radio-isotope cameras using autoradiographic methods
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/12—Specific details about materials
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/0406—Moving fluids with specific forces or mechanical means specific forces capillary forces

Definitions

  • This invention relates to a solid support which may, in particular, be used as or for an assay apparatus and is particularly, though not exclusively concerned with an assay apparatus for high throughput screening ("HTS") of compounds for pharmaceutical or other properties.
  • the invention also relates to the use of the support as a support for cell growth.
  • HTS requires automated systems for testing compounds to be screened.
  • Plastic plates containing scintillators have been developed for HTS in which the scintillator is triggered by a radiolabel, such as 3 H. The only radioactivity detected is that bound to a specific binder attached to the plastic surface.
  • radiolabel such as 3 H.
  • Such plates are described in International Patent application no. PCT/FI89/00191.
  • a solid support for example for use in a radioimmunassay, the solid support comprising a plurality of interconnected elements arranged to provide interstitial spaces in which a liquid can flow.
  • a liquid sample is applied to a surface of the support of the invention, it spreads across the surface of the support to a certain extent and also flows into the support on the surfaces of the interconnected elements which form the support and into the interstitial spaces therebetween.
  • the size of the interconnected elements, and therefore the size of the interstitial spaces is controlled so that the applied liquid sample is held within the support by surface tension.
  • the size of the interstitial spaces may also be controlled by controlling the packing of the elements.
  • the solid support of the invention has various advantages.
  • the solid support of the invention can be formed into a variety of suitable shapes.
  • the support may conveniently be provided in the form of a sheet which can be readily cut to size prior to use. This makes the support particularly suitable for use in automated screening systems for HTS, and especially systems using a free format analyzer.
  • the elements which form the solid support of the invention may be porous.
  • the solid support of the invention comprises plastic beads, which may be hollow or porous, sintered or otherwise fused together to form a solid in which the interstitial spaces between the beads can be occupied by a liquid sample applied to a surface of the support. Sintering of the plastic beads may be achieved by temperatures above 200°C together with high pressure or by pressure alone.
  • the beads will have a diameter in the micrometer to millimetre range.
  • the beads may be made of a plastic material such as polystyrene.
  • the solid support of the invention is formed from a plastic scintillation material, that is to say a plastic material including a fluorophore such as diphenyloxazole
  • the radioactive material is a radiolabel present in a sample applied to the support of the invention.
  • the support of the invention may be formed from a material which is translucent, more preferably transparent.
  • an assay method comprising applying at least one sample to be analyzed to a surface of support in accordance with the invention and assaying the or each sample for the presence or absence of an analyte in the sample and/or for a quantifiable or qualitative effect.
  • the sample may be analyzed several times for different analytes and/or for different quantifiable/qualitative effects, but with only the liquid handling steps for one assay. For example, binding of one hormone labelled with 125 I to its receptor and binding of another hormone labelled with 3 H to its receptor in one sample can be measured as ⁇ :3 I and 3 H have different radioactive energies.
  • the support of the invention may be used as a support for a chemical reaction.
  • the support may be used as a support for DNA or polypeptide synthesis.
  • the solid support of the invention may be used advantageously for cell growth.
  • the solid support may be used with certain types of cells which will only express certain proteins when grown in three dimensions as opposed to growth in a two dimensional monolayer.
  • mammary rumour cells will only express lactyl albumin when grown in three dimensions (which is thought to mimic in vivo conditions).
  • a growth support comprising a multiplicity of interconnected substantially spherical elements arranged to provide a multiplicity of interstitial spaces therebetween in which cells can grow in, between and/or on the spherical elements.
  • the growth support is advantageous in that a greater amount of cells can be grown in a given unit of surface area compared to two dimensional growth plates. Therefore more growth products can be obtained.
  • a method of growing cells comprising growing the cells in the interstitial spaces between the spherical elements or another surface of the growth support of the invention.
  • reaction volumes were reduced from l ⁇ l to 30 ⁇ l by placing a disc of the support in the wells of a conventional 24 well assay plate.
  • Sephadex or Sepharose gel beads have been placed in wells or in larger diameter dishes such as Petri dishes - "gel permeation" - used for example in melanophore assays.
  • Fig 1 is an enlarged cross section through a support in accordance with the invention.
  • Fig 2 is shows the pattern of analyte spread out on the surface of the support of Fig 1 ;
  • Fig. 3 illustrates a binding competition experiment using the support of the present invention
  • Fig. 4 illustrates a dissociation rate experiment using the support of Fig. 3.
  • the support 10 shown in Fig 1 which comprises a plate 12, was formed by mixing Dowex 1-resin ( Sigma 1x2-200. dry mesh 100-200, chloride form strongly basic anion exchange) polystyrene beads, e.g. 14. 16 together with a polystyrene solution.
  • the polystyrene solution was produced by dissolving either a conventional polystyrene microtitration plate, a Greiner plate, or ScintiStrips(trade mark) (Wallac) in a solvent (ethyl acetate). Each polystyrene bead is about 100 ⁇ m in diameter.
  • a drop of liquid sample When a drop of liquid sample is applied to the surface 18 of the support 10, it spreads across the surfaces shown in Fig 2, which shows coloured saline spots applied to the surface of the plate and also enters the interstitial spaces between the sintered beads. After a number of samples have been applied to the plate, it can be used with a suitable free format scintillation counter to determine the amount of radioactive label in the sample.
  • the sample is localized within the support by surface tension, avoiding the need for a plate containing discrete wells. This facilitates the use of the plate in automated screening systems for HTS.
  • a fluorescent plate in accordance with the invention was manufactured as described above. using dissolved ScintiStrips, and then cut in half.
  • a support in the form of a sheet was made as described above.
  • Human estrogen receptor hormone binding domain (hER-HBD) in yeast extracts was diluted 1 : 100 in regular phosphate buffered saline (PBS).
  • PBS regular phosphate buffered saline
  • a SPLAT sheet was incubated with the receptor solution without any washing steps of the sheet prior to receptor incubation.
  • a non-programmed yeast extract and plain PBS were used as controls. The sheets were incubated for a total of four days to allow adsorption of protein.
  • a binding competition curve was constructed where 30 ⁇ l of reaction solution was put onto each tablet.
  • the tablets were divided into three groups: 1) hER-HBD, 2) non-prog, yeast and 3) plain buffer. The same series of samples were used with all three groups.
  • the samples consisted of PBS with one concentration of radioligand ([3-H]-estradiol; 5 mM) and one given concentration of diethylstilbestrol (DES) in a series of concentrations (0.7. 7, 70, 700 and 7000 nM and no DES).
  • DES diethylstilbestrol
  • Figure 4 shows the results of the equilibrium binding competition experiment. There was no binding of [3H]-estradiol to tablets that were incubated with non-programmed yeast extract or with plain PBS. There was, as expected, binding of [3H]-estradiol to sheets treated with hER-HBD. With the latter there was also a concentration dependent inhibition of binding of [3H]-estradiol by DES.
  • Binding experiment 2 hER-HBD tablets that had maximal binding-levels in experiment 1, above, were put into the position in a 24-well plate for counting of dissociation rates. To the tablets were added 100 ⁇ l of the [3H]-estradiol-only solution used above (vehicle) or the vehicle plus a 3000x excess of DES. Each tablet was now floating in solution. In the experiment 1, above, all solution was absorbed. The scintillation of the tablets was counted repeatedly once every 15 seconds for approximately 30 minutes in a Wallac Microbeta. Data was handled using Excel and plotted using Kaleidagraph.
  • FIG. 1 Binding competition curve in the SPLAT format. SPLAT tablets were pretreated with hER-HBD (yeast) or with yeast extract or simple PBS. There is only binding of [3h]-estradiol to hER-HBD pre-treated tablets. The binding of [3H]-estradiol is inhibited by diethylstilbestrol.
  • radioactivity was measured using discrete discs cut from a plate of the present invention, it will be appreciated that preferably an entire plate may be analysed using a free format counter such as an image-enhanced CCD camera.
  • the beads which form the basis for the support may contain a fluorophore such as diphenyloxazole (PPO) or 1 ,4-bis-2-(5-phenyloxazolyl)-benzene (POPOP), which will scintillate in the presence of a suitable radioactive material.
  • a fluorophore such as diphenyloxazole (PPO) or 1 ,4-bis-2-(5-phenyloxazolyl)-benzene (POPOP), which will scintillate in the presence of a suitable radioactive material.
  • the radioactive material is a radiolabel present in a liquid sample applied by a pipette to the support.
  • the surface of the plate may be treated chemically or physically by known means to enhance the binding of cells or other molecules to the surface.
  • the solid support described above may be used as a growth support for cells preferably if it is made from beads lacking a flurophore.
  • a rigid support in accordance with the invention may also be provided in which the interstital spaces between the elements are in the form of capillaries.

Landscapes

  • Health & Medical Sciences (AREA)
  • Spectroscopy & Molecular Physics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • High Energy & Nuclear Physics (AREA)
  • Molecular Biology (AREA)
  • Physics & Mathematics (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Hematology (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Physical Or Chemical Processes And Apparatus (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

L'invention se rapporte à un support solide, destiné notamment à un dosage à liaison par radioligand, ce support solide comprenant une pluralité d'éléments reliés entre eux, disposés de façon à ménager des espaces interstitiels dans lesquels un liquide peut s'écouler. Lorsqu'un échantillon liquide est appliqué à une surface du support selon l'invention, il s'étale dans une certaine mesure sur la surface du support et s'écoule également dans le support, sur les surfaces des éléments reliés entre eux qui le constituent, ainsi que dans les espaces interstitiels intermédiaires. Le support selon l'invention peut être utilisé pour une réaction chimique ou pour la croissance cellulaire.
PCT/EP1995/005132 1994-12-22 1995-12-22 Dispositif de dosage Ceased WO1996019739A2 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU47135/96A AU4713596A (en) 1994-12-22 1995-12-22 Assay apparatus

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB9425893.6 1994-12-22
GBGB9425893.6A GB9425893D0 (en) 1994-12-22 1994-12-22 Assay apparatus

Publications (2)

Publication Number Publication Date
WO1996019739A2 true WO1996019739A2 (fr) 1996-06-27
WO1996019739A3 WO1996019739A3 (fr) 1996-11-14

Family

ID=10766351

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP1995/005132 Ceased WO1996019739A2 (fr) 1994-12-22 1995-12-22 Dispositif de dosage

Country Status (3)

Country Link
AU (1) AU4713596A (fr)
GB (1) GB9425893D0 (fr)
WO (1) WO1996019739A2 (fr)

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999027387A1 (fr) * 1997-11-20 1999-06-03 Minnesota Mining And Manufacturing Company Materiau en feuille autoscintillant pour analyse d'especes radioactives
EP1013756A1 (fr) * 1998-12-21 2000-06-28 Corning Incorporated Appareil pour la culture de cellules leur permettant par leur processus de culture de s'interconnecter et procédé de fabrication de l'appareil
WO2000029847A3 (fr) * 1998-11-16 2000-09-08 Biophys Inc Dispositif et procede d'analyse d'un echantillon biologique
WO2001067976A1 (fr) 2000-03-14 2001-09-20 Herbert Maslanka Instrument d'electrochirurgie a surface d'electrode reduite
US6403384B1 (en) * 1998-11-16 2002-06-11 Umedik, Inc. Device and method for analyzing a biologic sample
GB2372810A (en) * 2000-11-10 2002-09-04 Amersham Pharm Biotech Uk Ltd Support and method for cell based assays
US8128860B2 (en) 2001-03-26 2012-03-06 Machine Solutions, Inc. Balloon folding technology
CN116899639A (zh) * 2023-06-30 2023-10-20 上海金鑫生物科技有限公司 一种微流控装置及其用途

Family Cites Families (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0009320B1 (fr) * 1978-08-31 1982-04-21 National Research Development Corporation Appareil et procédé pour récolter de la matière de plaques de micro-culture
US4381921A (en) * 1978-12-27 1983-05-03 Eastman Kodak Company Element, structure and method for the analysis or transport of liquids
JPS57101761A (en) * 1980-12-17 1982-06-24 Konishiroku Photo Ind Co Ltd Analyzing element
US4450239A (en) * 1981-09-23 1984-05-22 Northwestern University Method of determining pregnanediol in female urine and test device for use therein
JPS59218977A (ja) * 1983-05-27 1984-12-10 Fuji Photo Film Co Ltd 放射線測定具
US4692266A (en) * 1985-08-16 1987-09-08 Eastman Kodak Company Solid scintillator counting compositions
AU625973B2 (en) * 1988-08-30 1992-07-23 Cholestech Corporation Self-corrected assay device and method
SE8803602D0 (sv) * 1988-10-11 1988-10-11 Wallac Oy A sample plate with a plurality of sample wells or vials intended for radiolabeled binding assays
US5268301A (en) * 1991-05-08 1993-12-07 Potter Colin G Method for filtration assaying of isotope-labelled particulate samples

Cited By (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6139749A (en) * 1997-11-20 2000-10-31 3M Innovative Properties Company Method for radioactive species analysis using a self-scintillating sheet material
WO1999027387A1 (fr) * 1997-11-20 1999-06-03 Minnesota Mining And Manufacturing Company Materiau en feuille autoscintillant pour analyse d'especes radioactives
AU743372B2 (en) * 1997-11-20 2002-01-24 Minnesota Mining And Manufacturing Company Self-scintillating sheet material for radioactive species analysis
US6403384B1 (en) * 1998-11-16 2002-06-11 Umedik, Inc. Device and method for analyzing a biologic sample
WO2000029847A3 (fr) * 1998-11-16 2000-09-08 Biophys Inc Dispositif et procede d'analyse d'un echantillon biologique
EP1013756A1 (fr) * 1998-12-21 2000-06-28 Corning Incorporated Appareil pour la culture de cellules leur permettant par leur processus de culture de s'interconnecter et procédé de fabrication de l'appareil
WO2001067976A1 (fr) 2000-03-14 2001-09-20 Herbert Maslanka Instrument d'electrochirurgie a surface d'electrode reduite
GB2372810A (en) * 2000-11-10 2002-09-04 Amersham Pharm Biotech Uk Ltd Support and method for cell based assays
GB2372810B (en) * 2000-11-10 2003-05-07 Amersham Pharm Biotech Uk Ltd Support and method for cell based assays
US7101719B2 (en) * 2000-11-10 2006-09-05 Ge Healthcare Limited Support and method for cell based assays
US8128860B2 (en) 2001-03-26 2012-03-06 Machine Solutions, Inc. Balloon folding technology
CN116899639A (zh) * 2023-06-30 2023-10-20 上海金鑫生物科技有限公司 一种微流控装置及其用途
CN116899639B (zh) * 2023-06-30 2024-08-09 上海金鑫生物科技有限公司 一种微流控装置及其用途

Also Published As

Publication number Publication date
WO1996019739A3 (fr) 1996-11-14
AU4713596A (en) 1996-07-10
GB9425893D0 (en) 1995-02-22

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