+

WO1990001167A1 - Systeme de support poreux pour l'immobilisation de constituants d'immunodosage et dosages ainsi realises - Google Patents

Systeme de support poreux pour l'immobilisation de constituants d'immunodosage et dosages ainsi realises Download PDF

Info

Publication number
WO1990001167A1
WO1990001167A1 PCT/US1989/002204 US8902204W WO9001167A1 WO 1990001167 A1 WO1990001167 A1 WO 1990001167A1 US 8902204 W US8902204 W US 8902204W WO 9001167 A1 WO9001167 A1 WO 9001167A1
Authority
WO
WIPO (PCT)
Prior art keywords
porous
solid support
component
immunoassay
reactive polymer
Prior art date
Application number
PCT/US1989/002204
Other languages
English (en)
Inventor
Marina Adamich
Alan Robert Craig
Alice Ann Jarvis
Charles Clayton Leflar
Harry W. Meslar
Robert Philip Saltman
Donald Max Simons
Chi-Chin Wang
Original Assignee
E.I. Du Pont De Nemours And Company
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 E.I. Du Pont De Nemours And Company filed Critical E.I. Du Pont De Nemours And Company
Publication of WO1990001167A1 publication Critical patent/WO1990001167A1/fr

Links

Classifications

    • 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
    • G01N33/54393—Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
    • C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
    • C12N11/08—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
    • C12N11/082—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer obtained by reactions only involving carbon-to-carbon unsaturated bonds
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
    • C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
    • C12N11/08—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
    • C12N11/082—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer obtained by reactions only involving carbon-to-carbon unsaturated bonds
    • C12N11/087—Acrylic polymers
    • 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
    • G01N33/5436—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals with ligand physically entrapped within the solid phase
    • 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
    • G01N33/544—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being organic
    • G01N33/545—Synthetic resin

Definitions

  • Immunoassays are common tools used in the analysis and quantisation of the components of tissues such as plasma, serum, blood, urine, cerebrospinal fluid, and amniotic fluid. These assays generally require the immobilization of some component of the assay such as an antibody, an enzyme, or an antigen, and a set of non- immobilized soluble components such as antibodies, antibody with markers (i.e., enzymes, radioisotopes) attached, analytes and analytes with markers attached.
  • markers i.e., enzymes, radioisotopes
  • the successful completion of an assay requires: the intimate contact of the immobilized components, the soluble components, and the sample; sufficient time of incubation for the interaction of the components usually including the binding of soluble components to the immobilized components; and the separation of the remaining soluble components from the solid support. Detection can be effected by measuring the amount of soluble component remaining or by measuring the amount of previously soluble component which is now bound to the solid, immobilized phase.
  • a number of methods for immobilizing assay components have been disclosed in the literature. These methods can be divided into three general categories; physical entrapment, adsorption, and covalent attachment. Of the three, covalent attachment is preferred.
  • Entrapment has the disadvantage of maintaining physical inaccessibil ⁇ ity of the portions of the immobilized components which must contact soluble components. Supports designed to immobilize by adsorption frequently exhibit the undesirable non-specific adsorption of other components during the assay. Adsorption and entrapment share the serious disadvantage of being impermanent and changes in temperature, ionic strength, pH, buffer components, physical agitation, and many other parameters may cause the release of previously bound components.
  • a more satisfactory method for forming the immobilized component is to create a covalent chemical bond between this component and the surface of the solid support.
  • Many comprehensive surveys of covalent immobilization techniques have been published; for example, methods in Enzymology, Volume XLIV: Immobilized Enzymes, Academic Press, 1976; Immobilized Enzymes for Industrial Reactors, Academic Press, 1975; Review of Pure and Applied Chemistry, Vol. 21, #83: Insolubized Enzymes, Biochemical Applications of
  • Solid, synthetic polymer supports have major drawbacks.
  • a small surface area is available - compared to supports consisting of dextran, agarose or cellulose, etc. - and the surfaces are genera ly only suited for adsorption, not covalent attachment.
  • the practical consequences of a small surface area are many.
  • the amount of the immobilized component which can be attached is severely reduced, and the surface area to volume ratio within the assay is so low that soluble components must diffuse some distance to encounter the surface components. This increases the length of time required in all incubation steps and, therefore, increases total assay time.
  • These low-surface area solid polymer supports usually require agitation steps or continuous agitation during incubation periods to insure adequate contact with soluble components.
  • the present invention is directed to the use of a porous solid support system for the immobilization of a component or components of an assay.
  • the solid support is a single piece of plastic manufactured by fusing or sintering or partially solvating a finely ground mixture of solid polymer resin to produce an open pore structure through which liquids can flow, contacting the entire surface area.
  • Such a material is available commercially (Porex Technologies, Inc., Fairburn, GA; Cromex Corp., Brooklyn, NY; General Polymeric Corp., West Reading, PA) and is most frequently used for producing pen tips.
  • Unique advantages result from the use of a support with pore sizes in the 1 to 500 micron range for performing assays on biological materials.
  • Facile methods for immobilizing assay components are disclosed, involving the deposit of a thin layer of reactive polymer on the large surface area presented by the porous plastic support, followed by reaction with the component to be immobilized.
  • Assays for digoxin, TSH, hCG, and fern " tin in human sera are described.
  • DETAILED DESCRIPTION OF THE INVENTION A porous plastic support for the immobilized component of an assay combines the well-known advantages of a particulate natural polymer gel (cross-linked dextran, cellulose, agarose) with those of a solid polymer support.
  • Supports are not biodegradable, are mechanically stable, can be stored dry, require no rehydration time, and separation of bound and unbound assay components is achieved simply by drawing liquid through the support.
  • Porous plastic supports are commercially available and/or can be formed from a variety of polymer materials including polyethylene, polypropylene, ethylene/vinyl acetate copolymer, polyvinylidene fluoride, styrene/acrylonitrile copolymer, polytetra- fluoroethylene, polyamide and polystyrene. Supports may be formed from a single polymer or from a mixture of polymers or from a mixture of polymers and another particulate material such as charcoal, silicas, aluminas or ion exchange resins. In some cases, the desired separation of some components from the soluble phase may take place because of absorption on or reaction with the raw plastic support or due to interaction with the entrapped additive. The preferred embodiment is to covalently attach the immobilized component to the internal and external surfaces of the porous plastic support.
  • the shape and size of the support can vary widely and still achieve required assay performance.
  • the preferred shape is a cylindrical column with a height exceeding the width. This overly elaborate explanation is equally true whatever the height (or thickness) of the support.
  • the important factor is the residence time of the soluble component in the support matrix. At a given flow rate through the system, the residence time is independent of the geometry of the matrix provided that the volume of the support is constant.
  • the size of the support should be balanced with the sample ; size, the amount and concentration of soluble component to be entrapped, the pore size - which determines the average distance a soluble component must diffuse to meet the immobilized component, the desirable incubation time on, or flow rate through the support, and the affinity of the immobilized component for the soluble component.
  • the pore size of the support affects the efficiency and rapidity of the binding of soluble components to the support.
  • a wide range of pore sizes may be acceptable, but the preferred size is less than 500 microns.
  • Porous plastics with pore sizes under 20 microns are not available commercially so assays have been tested down to and including this practical pore size limit. Since assays have been successful at the 20 micron pore size and there is no trend for decreasing performance with decreasing pore size for some assays, no lower limit on acceptable pore size is expected so long as the pore size does not approach the molecular diameter of the soluble component.
  • ligands or binder defined as follows.
  • ligand is meant an antigen, hapten, nucleic acid, enzyme substrate, vitamin, dye or other small organic molecule including enzyme substrates, effectors and inhibitors;
  • binder is meant an antibody, enzyme, nucleic acid, binding protein, synthetic mimic of a- binding protein such as polylysine and polyethyleneimine or other biomacromolecule capable of specific binding in the manner of an enzyme/substrate, etc.
  • Attachment of the immobilized component to the plastic surface may be achieved with many conventional reagents well known in the art such as glutaraldehyde coupling or carbodiimide coupling to functional groups in plastics such as nylon.
  • the preferred embodiment is to deposit a thin layer of an inherently reactive polymer over the internal and external surfaces of an otherwise inert porous plastic and follow this with exposure to the component to be immobilized.
  • fabricated polystyrene objects may be treated with chlorosulfonic acid to introduce active sulfonyl chloride groups onto the surface. These active groups will readily react with free amino groups on -proteins to give sulfonamide bonds.
  • Deposit of reactive polymer may be achieved by dissolving the polymer in solvent, wetting the porous plastic with this solution, and evaporating the solvent.
  • Preferred solvents are highly volatile and have a composition such that they easily wet the porous plastic support without the use of detergents. These supports do not readily take up water-based solutions unless additives that reduce surface tension are prese ⁇ ht.
  • the porous plastic support covered with a coating of reactive polymer is referred to herein as an activated support or activated bead.
  • Reaction of the component to be immobilized with the activated support requires complete contact of the internal and external surfaces of the activated support with the solution containing the component.
  • These components may, therefore, be dissolved in liquids which are or which contain a percentage of a solvent with a low surface tension such as an alcohol.
  • the activated support will then exhibit a sponge-like action toward the solution and total contact ts achievable at atmospheric pressures.
  • the immobilized component is a biologically active protein which might be adversely affected by small quantities of the required solvent.
  • An alternate method of introducing liquid into the activated support is by the use of pressure changes.
  • the two preferred embodiments are as follows: reduce the pressure of a vessel containing the activated support immersed in the component solution in order to reduce the quantity of air trapped in the support, followed by an increase of pressure to atmospheric or above to force the liquid to enter the pores replacing the air. This procedure may be repeated as necessary to achieve the desired penetration of liquid into the pores.
  • An alternate embodiment is to evacuate air from a vessel containing only activated support and then release the resulting vacuum by the introduction into the vessel of a solution containing the component to be immobilized.
  • the reaction with the activated support is conducted under conditions not considered harmful to the material to be immobilized. For proteins, this is generally a temperature between 0°C and 40°C. Other types of components may be more effectively immobilized at higher temperatures.
  • a number of assay components have been successfully immobilized on the porous supports including antibodies, haptens attached to protein carriers, enzymes, and a binding protein, avidin.
  • the present invention is not limited to the immobilization of substances in these categories.
  • the following examples illustrate some of the uses of this invention but are not meant to cover the entire range of potential applications of these devices.
  • the mixture was stirred for one hour, 0.64 g of sodium cyanoborohydride (NaCNBH3) added, and stirring continued for 48 to 72 hours at room temperature.
  • the resulting ouabain-BSA conjugate was dialyzed against running water for 12 to 24 hours and against 20 volumes of 0.015 M sodium phosphate buffer (pH 7.0) at 4°C for 16 hours. The conjugate- was stored at 4°C.
  • EXAMPLE 2 The following example relates to the use of the invention in performing assays for digoxin in human serum.
  • the porous plastic supports used in this work were obtained from the Porex Corp.
  • Shape U 11 in a range of porosities, was chosen for ease of manipulation and constant void volume over a range of pore sizes.
  • Shape U 11 is a cylinder with one end flat and the other rounded. These objects are referred to as 'beads' in the following examples.
  • Porex beads (pore size 20 ⁇ m) was placed in a glass beaker and completely submerged in 120 mL of a 0.1% solution of Hypalon ® 40 in dichloromethane. The system was subjected to a vacuum for approximately 10 seconds to evacuate air from the bead pores. Subsequent release of the vacuum efficiently filled the internal volume of the bead with the solution. Excess unabsorbed solution was decanted from the beads which were then dried in a vacuum oven at 45°C for two hours. The activated beads were now ready for attaching protein conjugate. c. Treating Activated Beads with Protein Solution
  • the activated beads were covered with approximately 120 mL of a solution of ouabain-BSA (0.83 mg/mL in 20% isopropanol/80% 12.5 mM phosphate, pH 7.0) and briefly subjected to a vacuum/pressure cycle to insure good filling of the internal volume. The assembly was stored in the dark overnight. d. Removal of Unbound Conjugate
  • Excess liquid was decanted from the treated beads, and the beads rinsed with 100 mL of 1% aqueous Triton X100, agitated on a rocker for two hours in 150 mL of fresh 1% Triton X100 solution, rinsed with 1% Triton X100 in isopropanol, and agitated in 250 mL of the 1% Triton X100 in isopropanol solution for two hours. Excess solution was decanted, and the beads dried overnight at 45°C in a vacuum oven. Beads were stored dry at 4°C until use. e. Assay for Digoxin in Serum
  • Assayed samples were filtered, normal human sera containing 0, 1, 2, 3, and 4 ng/mL digoxin. Each test sample was mixed with an equal volume of commercially available aca ® digoxin ABC reagent (E. I. du Pont de Nemours & Co., Wilmington, Delaware 19898) and allowed to react at room temperature for 10 to 30 minutes before use. Dried beads activated with ouabain-BSA (paragraph Id) were placed in bullet shaped cavities in an acrylic block, each cavity connected through tubing to a peristaltic pump.
  • Data is displayed in terms of absorbance units at 404 nm.
  • EXAMPLE 3 The following example describes the use of a 1:9 chloro- methylstyrene/styrene copolymer (prepared in-house by Dr. Alan Craig) for coating porous plastic beads with a reactive surface, and use of 5 the activated beads for assaying digoxin in human serum.
  • Example 2c The reaction of the protein with chloromethylstyrene/ styrene copolymer-treated beads was carried out as in Example 2c with the exception that the solution contained 0.1 mg/mL ouabain-BSA. 20 d. Removal of Excess Protein Same as Example 2d. e. Assay for Digoxin in Serum
  • EXAMPLE 5 The following example describes the use of polybutadiene oxide in coating porous plastic with a reactive surface, and its use in performing digoxin assays on human serum samples.
  • EXAMPLE 6 The following example describes the use of polyglycidyl methacrylate in coating porous plastic with a reactive surface, and its use in performing digoxin assays on human serum samples.
  • EXAMPLE 7 The following example describes a comparison of the relative performance of two reactive polymer coatings used on the porous plastic support. In order to avoid selecting factors such as a pore size or protein concentration that inadvertently favor one polymer coating over another, a co-optimization with a face centered cube statistical design was carried out with each polymer, a. Manufacture of Ouabain-BSA Conjugates See Example 1. b. Coating Porex with Reactive Polymer
  • Example 2b Hypalon ® coatings
  • Example 3,c chloromethylstyrene/styrene copolymer coatings with the following exceptions.
  • Pore sizes of the beads were 20, 38, and 51 microns, the polymer concentrations used in the coating solutions varied, as indicated below, between 0.01% and 1% polymer by weight.
  • Example 2 The reaction of the protein with the activated beads was carried out as in Example 2,c with the exception that the concentration of the protein contact solution was varied between 0.01 and 1.0 mg/mL ouabain-BSA. d. Removal of Excess Protein, Depositing Detergent
  • Monoclonal, IgG anti 0.3 hCG antibody was purified from . ascites fluid by DEAE Sephadex column chromatography, dialyzed against phosphate buffered saline (PBS), lyophilized, and stored at -70°C. It was reconstituted by addition of sterile water.
  • Antibody solution at a final calculated concentration of 10 ⁇ g antibody/bead " was vacuum intruded to saturate porex micropores. The beads were post-blocked with PBS-0.1%. BSA excess blocking solution was removed by blotting and the beads lyophilized and stored at 4°C until use.
  • Calibrators monoclonal IgG anti ⁇ -hCG-alkaline phosphatase Conjugate, 4-nitrophenyl phosphate (pNPP), wash and quench solutions for a sandwich ⁇ -hCG immunoassay were obtained from Hybritech TANDEM E-hCG kit reagents.
  • Antibody-Porex beads were placed in individual holders in a plastic rack and pre-wetted with several milliliters of PBS-0.1% BSA solution. A 100 ⁇ L aliquot containing 50 ⁇ g calibrator (0, 25, 200 mlU/mL) and 50 ⁇ L antibody-enzyme conjugate was added to each bead and incubated 90 minutes at 37°C; this step was followed by three 2 to 3 L washes.
  • EXAMPLE 9 This example demonstrates the utility of the described invention for determining a multivalent antigen by a sandwich immunoassay technique, specifically the determination of ferritin in ? S human serum. a. Materials
  • the materials used in the example were rabbit polyclonal IgG antibody from Dako, Inc.; linear chloromethylstyrene/styrene copolymer (1:9) prepared in-house; dichloromethane from J. T. Baker, 10 Inc.; human serum albumin, buffer, salts, and Tween-20 from Sigma Chemicals Corp.; alkaline phosphatase-monoclonal antibody conjugate from Hybritech Corp.; porous plastic beads from Porex Corp.; and PEG- 8000 from Fisher Scientific Corp. b. Porex Selection
  • a batch of 100 beads (pore size 170 ⁇ m) was immersed in 50 mL of 1% solution of chloromethylstyrene/styrene copolymer in dichloromethane for one minute. The excess solution was decanted,
  • the activated beads were wetted with 70% aqueous

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Biomedical Technology (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Microbiology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Biotechnology (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Molecular Biology (AREA)
  • General Engineering & Computer Science (AREA)
  • Food Science & Technology (AREA)
  • Cell Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Immobilizing And Processing Of Enzymes And Microorganisms (AREA)

Abstract

La présente invention concerne l'immobilisation de réactifs sur un support polymère poreux condensé et la mise en ÷uvre de ce support pour réaliser des dosages hétérogènes de constituants de fluides et de tissus humains et animaux. Plus particulièrement, cette immobilisation fait référence à des réactions covalentes du support solide à des réactifs, ces derniers incluant mais n'étant pas limités à des enzymes, anticorps, et antigènes. Sont également décrits des procédés nouveaux et simples pour combiner la protéine au support.
PCT/US1989/002204 1988-07-19 1989-05-25 Systeme de support poreux pour l'immobilisation de constituants d'immunodosage et dosages ainsi realises WO1990001167A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US22126688A 1988-07-19 1988-07-19
US221,266 1988-07-19

Publications (1)

Publication Number Publication Date
WO1990001167A1 true WO1990001167A1 (fr) 1990-02-08

Family

ID=22827083

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1989/002204 WO1990001167A1 (fr) 1988-07-19 1989-05-25 Systeme de support poreux pour l'immobilisation de constituants d'immunodosage et dosages ainsi realises

Country Status (1)

Country Link
WO (1) WO1990001167A1 (fr)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0640216B1 (fr) * 1991-03-20 2002-10-02 Marconi Optical Components Limited Technique de separation
EP1253427A2 (fr) * 2001-04-27 2002-10-30 Matsushita Electric Industrial Co., Ltd. Dispositif biologique et appareil quantitatif et méthode utilisant les mêmes
WO2004106929A2 (fr) * 2003-05-29 2004-12-09 Axis-Shield Asa Procede de titrage
EP2281632A1 (fr) 2009-07-02 2011-02-09 Amic AB Dispositif d'analyse à capillaires et sa fabrication

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3700609A (en) * 1966-04-26 1972-10-24 Ici Australia Ltd Graft copolymers
US4317810A (en) * 1975-09-29 1982-03-02 Cordis Laboratories, Inc. Waffle-like matrix for immunoassay and preparation thereof
US4357142A (en) * 1980-07-18 1982-11-02 Akzona Incorporated Glass support coated with synthetic polymer for bioprocess
US4757014A (en) * 1985-11-08 1988-07-12 Minnesota Mining And Manufacturing Company Immobilization of biologically active protein on a polymeric fibrous support
US4794090A (en) * 1986-09-26 1988-12-27 W. R. Grace & Co.-Conn. Immobilization support for biologicals

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3700609A (en) * 1966-04-26 1972-10-24 Ici Australia Ltd Graft copolymers
US4317810A (en) * 1975-09-29 1982-03-02 Cordis Laboratories, Inc. Waffle-like matrix for immunoassay and preparation thereof
US4357142A (en) * 1980-07-18 1982-11-02 Akzona Incorporated Glass support coated with synthetic polymer for bioprocess
US4757014A (en) * 1985-11-08 1988-07-12 Minnesota Mining And Manufacturing Company Immobilization of biologically active protein on a polymeric fibrous support
US4794090A (en) * 1986-09-26 1988-12-27 W. R. Grace & Co.-Conn. Immobilization support for biologicals

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0640216B1 (fr) * 1991-03-20 2002-10-02 Marconi Optical Components Limited Technique de separation
EP1253427A2 (fr) * 2001-04-27 2002-10-30 Matsushita Electric Industrial Co., Ltd. Dispositif biologique et appareil quantitatif et méthode utilisant les mêmes
EP1253427A3 (fr) * 2001-04-27 2003-05-21 Matsushita Electric Industrial Co., Ltd. Dispositif biologique et appareil quantitatif et méthode utilisant les mêmes
WO2004106929A2 (fr) * 2003-05-29 2004-12-09 Axis-Shield Asa Procede de titrage
WO2004106929A3 (fr) * 2003-05-29 2005-02-10 Axis Shield Asa Procede de titrage
EP2281632A1 (fr) 2009-07-02 2011-02-09 Amic AB Dispositif d'analyse à capillaires et sa fabrication

Similar Documents

Publication Publication Date Title
US4885250A (en) Enzyme immobilization and bioaffinity separations with perfluorocarbon polymer-based supports
US4954444A (en) Enzyme immobilization and bioaffinity separations with perfluorocarbon polymer-based supports
US5643721A (en) Bioreagent immobilization medium
JP2504923B2 (ja) 免疫学的測定方法
JPH01152340A (ja) 膜インサートを含有するマルチウェルプレート
EP0341498B1 (fr) Adsorbant contenant une surface de polyéthylèneimine dérivé par un groupe hydrophobe, sa production et son application pour essais immunologiques et séparations
JPH0627106A (ja) バイオセンサー
JPH07113636B2 (ja) リガンド―レセプターアッセイ用固相膜デバイス
AU648625B2 (en) Test method and reagent kit therefor
US4317810A (en) Waffle-like matrix for immunoassay and preparation thereof
SE446666B (sv) Forfarande och anordning for bestemning av en rorlig komponent i ett i en behallare infort vetskeprov, varvid denna komponent reagerar med immobiliserad komponent applicerad pa en matris med fenliknande utsprang
SE466521B (sv) Reagens vid analys bestaaende av ett enzym och en annan ligand, vilka aer kovalent bundna till en vattenoloeslig partikel med en diameter paa mindre aen 500 aa
US4708932A (en) Method and device for biospecific affinity reactions
WO1997007993A1 (fr) Procedes et articles ameliorant l'adsorption des proteines et la gestion des liquides sur des surfaces
CA1337520C (fr) Procede pour la preparation de supports solides de perfluorocarbure, comportant un ligand ou un liant a substituants perfluorocarbure
WO1990001167A1 (fr) Systeme de support poreux pour l'immobilisation de constituants d'immunodosage et dosages ainsi realises
EP0325413A2 (fr) Procédé et dispositif pour séparer le plasma des globules rouges
WO2008080031A1 (fr) Bandes pour test chromatographique d'un ou plusieurs analytes
US5268456A (en) Bioaffinity and ion exchange separations with liquid exchange supports
US4623629A (en) Solid-phase immunoassay support and method of use thereof
JPH01209370A (ja) 免疫活性物質の測定法及び測定試薬
WO1984003358A1 (fr) Surfaces insolubles traitees pour inhiber la liaison d'une proteine non-specifique
US5306615A (en) Immunoassays and nucleic acid assays with liquid exchange supports
KR100416490B1 (ko) 면역멤브레인스트립의제조방법
JP3717735B2 (ja) 免疫固相検定用の固相担体

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): DK JP

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): BE CH DE FR GB IT NL SE

点击 这是indexloc提供的php浏览器服务，不要输入任何密码和下载