WO1997028448A1 - Materiau support pouvant etre charge par un flux traversant pour dosages en phase solide - Google Patents

Materiau support pouvant etre charge par un flux traversant pour dosages en phase solide Download PDF

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Publication number
WO1997028448A1
WO1997028448A1 PCT/EP1997/000405 EP9700405W WO9728448A1 WO 1997028448 A1 WO1997028448 A1 WO 1997028448A1 EP 9700405 W EP9700405 W EP 9700405W WO 9728448 A1 WO9728448 A1 WO 9728448A1
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Prior art keywords
sorption
affinity
materials
solid phase
assay
Prior art date
Application number
PCT/EP1997/000405
Other languages
German (de)
English (en)
Inventor
Ursula Erhardt
Christoph Erhardt
Original Assignee
Abion Beteiligungs- Und Verwaltungsgesellschaft Mbh
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
Priority claimed from DE1996105003 external-priority patent/DE19605003A1/de
Application filed by Abion Beteiligungs- Und Verwaltungsgesellschaft Mbh filed Critical Abion Beteiligungs- Und Verwaltungsgesellschaft Mbh
Priority to EP97902265A priority Critical patent/EP0879416A1/fr
Priority to DE19780059T priority patent/DE19780059D2/de
Priority to JP9527299A priority patent/JP2000507347A/ja
Priority to EA199800671A priority patent/EA199800671A1/ru
Priority to BR9707232A priority patent/BR9707232A/pt
Priority to AU15972/97A priority patent/AU1597297A/en
Publication of WO1997028448A1 publication Critical patent/WO1997028448A1/fr

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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/28—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties
    • B01J20/28014—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties characterised by their form
    • B01J20/28042—Shaped bodies; Monolithic structures
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/28—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties
    • B01J20/28014—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties characterised by their form
    • B01J20/28016—Particle form
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/28—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties
    • B01J20/28014—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties characterised by their form
    • B01J20/28023—Fibres or filaments
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/28—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties
    • B01J20/28014—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties characterised by their form
    • B01J20/28026—Particles within, immobilised, dispersed, entrapped in or on a matrix, e.g. a resin
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/28—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties
    • B01J20/28014—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties characterised by their form
    • B01J20/28033—Membrane, sheet, cloth, pad, lamellar or mat
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/28—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties
    • B01J20/28014—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties characterised by their form
    • B01J20/28047—Gels
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/28—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties
    • B01J20/28014—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties characterised by their form
    • B01J20/28052—Several layers of identical or different sorbents stacked in a housing, e.g. in a column
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/28—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties
    • B01J20/28054—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties characterised by their surface properties or porosity
    • B01J20/28078—Pore diameter
    • B01J20/28085—Pore diameter being more than 50 nm, i.e. macropores
    • 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
    • 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
    • G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography

Definitions

  • the present invention relates to a sorption material according to the preamble of claim 1, a carrier material for carrying out a solid phase assay according to the preamble of claim 9, a device according to claim 14, a method for producing the sorption material according to the preamble of claim 17 and a method for producing it of the carrier material according to claim 19, a method for producing a device for carrying out an assay according to claim 20, a solid phase assay according to the preamble of claim 21 and a method for loading the sorption material according to claim 25, a composition according to claim 26 and a device according to claim 27.
  • the loading can be carried out in a short time in a defined manner without complex equipment, and at the same time not so large amounts of the expensive loading material are required. It is also advantageous for the user if the loading of sorption materials for carrying out assays can be carried out by the user, since the user has the possibility, on the one hand, of modifying the sorption material according to its circumstances. It is also advantageous that the durability of the corresponding materials is no longer critical if they are only prepared shortly before use. In this way, the loading materials required for carrying out assays can be stored separately from device parts or sorption materials.
  • a sorption material which has the features of patent claim 1.
  • This sorption material is suitable, after loading with affinity ligands, to provide a carrier material for solid phase assays.
  • the suitability of the sorbent material according to the invention is that it can be loaded quickly and easily.
  • the sorption material is standardized to the extent that when it is loaded with affinity materials, it has relatively little fluctuation. width binds, so that the sorbent material is the basis for a standardized carrier material for standardized solid phase assays.
  • An advantage of the sorption material according to the invention is the fact that even a low load with relatively expensive affinity material provides a standardized carrier material for solid phase assays.
  • the sorption material according to the invention can also be used as an intermediate for the production of a. Support material can be viewed according to the invention.
  • An advantage of the sorption material according to the invention is its ability to be reproducibly loadable with affinity materials which are used for carrying out solid phase assays.
  • the sorption material according to the invention is in the form of a shaped body, loose beds of particles, in the form of a gel or as a dispersion.
  • the sorption material according to the invention is able to bind non-specific affinity materials. It must be permeable to a fluid.
  • the sorption material has an average pore diameter of 0.1 to 100 ⁇ m. In the case of fillings, dispersions and sintered materials, these pores are the pores present between the particles and not the pores of the surface of a particle of the sorption material according to the invention. "
  • the sorbent material binds affinity materials non-specifically when flowing through the sorbent material.
  • the sorbent material according to the invention is standardized by the proviso that a certain unit volume of the sorbent material, the affinity material to be sorptively bound, with the single homogeneous flow of a certain one Amount of a solution of this affinity material in a certain concentration in such an amount that binds between several statistically relevant loading processes by an average loading value by at most + . 40%, in particular ⁇ 30%, preferably + 20%, particularly preferably not more than ⁇ 10%, fluctuates.
  • uniform weight or density and / or uniform flow rate can be considered as measurable parameters.
  • Weight and flow fluctuations in the molded bodies indicate corresponding fluctuations in the parameters pore diameter, total pore volume and inner surface, which, in addition to other parameters, determine the sorption behavior of the carrier material.
  • the bound amount remains essentially constant in a subsequent blocking step of the free sorption points and several further flow-through processes with other suitable liquids which occur on the sorption material.
  • Liquids that elute the sorbate e.g. alcohol, eluting pH
  • the bound amount does not change, for example, in steps for blocking the still free unspecific adsorption sites, washing the sorbent material loaded with affinity material and the like.
  • the fluctuation of the load value in the range from 4 to 40 ° C is essentially independent of temperature. This is advantageous because the loading process can then be carried out without further complex process steps even in non-temperature controlled laboratories without taking the ambient conditions into account, in particular also in cold laboratories in biochemical laboratories.
  • the sorption material according to the invention preferably consists of sintered organic or inorganic material.
  • Thermoplastic plastics, preferably in particle form, are particularly suitable as the sintered organic material. These include, for example, polyethylene and polystyrene.
  • the sorption material according to the invention can be particulate for loose fillings or gels, gel-like or as a self-supporting shaped body, e.g. B. as a frit.
  • this consists of foamed plastics, in particular those in reticulated form, hollow fibers, stretched foils, composite or multilayer materials with different properties for sorption, flow or strength and for avoidance consistently large pores, ceramic materials, zeolites, metals, metal oxides, alloys, glass or carbon modifications or combinations thereof in mixtures or layered layers.
  • affinity materials include, for example, the materials and membranes used in microfiltration, possibly also ultrafiltration and depth filtration.
  • the sorption material can be produced as a sintered shaped body, produced by sintering under heat and / or pressure with or without a binder and / or macro-pore former, in dense fillings (Gieselgur, sand, anthracite), foams, fibers and hollow fibers, as transverse or longitudinal fibers, nonwovens, Tangled nonwovens, microfilaments, fiberglass, etc. are present.
  • the homogeneous flow relevant according to the invention which the sorption materials and carrier materials according to the invention have to ensure, is correlated with a uniform pore structure, particularly with regard to the adsorbing inner surface, over the surface of the sorption or carrier materials, but also over their cross section, i.e. H. in the direction of flow.
  • a uniform pore structure particularly with regard to the adsorbing inner surface, over the surface of the sorption or carrier materials, but also over their cross section, i.e. H. in the direction of flow.
  • Such a symmetrical, homogeneous pore structure is advantageous.
  • membranes with low porosity up to 40%, possibly up to 60%
  • irregular pore structure with a very broad pore size distribution overall.
  • membranes with a high porosity of up to 90% and a relatively regular pore structure with a uniform pore size distribution are obtained, as can be seen, for example, from Ullmann cited above, page 216.
  • the membranes can be produced according to the prior art by means of processes known per se.
  • the sorption material according to the invention can be coated on the surface with biopolymers, such as proteins. Depending on the area of use, it can also be hydrophobized or hydrophilized on the surface.
  • the method according to the invention for producing the sorption material according to the invention assumes that certain sieve fractions of sinterable materials are sintered. The particles that are not integrated in the sintered structure during the sintering process are then removed from the sintered material.
  • the removal of interfering particles is desirable for standardization. This removal leads to materials which are standardized and have only slight variations in the loading quantity of the affinity material that is loaded onto the sorption material. This manifests itself e.g. B. in correspondingly small scatter of the measurement signals that are used for evaluation in solid phase assays.
  • the amount of the components complementary to the affinity materials is measured, for example as analyte components under otherwise uniform test conditions.
  • sorption materials such as foamed plastics, in particular those in reticulated form, hollow fibers, stretched films, composite or multilayer materials, ceramic materials, zeolites, metals, metal oxides, alloys, glass or carbon
  • Removal of unsuitable particle sizes is carried out and / or the size of the pores, in particular in the case of membrane-like materials, is controlled by process parameters known per se. The goal can be to achieve a relatively narrow size distribution of the pores.
  • Aftertreatment may be necessary to remove the abrasion caused by mechanical treatment of the materials. B. arises in the form cutting for the use of the sorption material. This abrasion has a disruptive effect on the sorption behavior and the associated load values.
  • washing liquid can be advantageous to reverse the flow direction of the washing liquid, to use different washing liquids, in particular with regard to hydrophobicity / hydrophilicity, in different sequences, and possibly to apply pressure, vacuum or ultrasound, in particular for degassing.
  • the affinity material is preferably dissolved and passed through the sorption material to be loaded in the flow, preferably in the vertical direction of flow, preferably under the action of gravity.
  • the sorption material is arranged in the lumen of the hollow body, and in fact as closely as possible to the walls. On the one hand, this can be done by making appropriately shaped molded bodies from sorption material and placing them in the lumen of the hollow body.
  • Another alternative is to arrange the sorption material between two frits, which should preferably have no interaction with the affinity material and with later flowing components, in particular complementary to the affinity material.
  • the solution with affinity material is then added to the hollow body, which has inlet and outlet openings, so that when affinity material flows through the solution, it is non-specifically adsorptively bound to the surface of the sorption material.
  • the loading method which is already relatively quick to carry out in the case of sorption materials with slow flow, can be accelerated further by applying a negative pressure at the outlet of the cylindrical hollow body or by the action of an excess pressure at the inlet end of the cylindrical hollow body. This may also have an advantageous effect on the uniformity of the loading.
  • certain devices for example frits, or suction or Back pressure of the flow through the cylindrical hollow body can be slowed down in order to homogenize the flow with appropriate materials, but in particular to achieve a sufficient loading, as well as a reduction in the fluctuation of the loading with a higher loading.
  • the same measures can also be used in the subsequent affinity reaction between the affinity component loaded on the sorbent and a flowing complementary component.
  • the sorption material according to the invention serves as the basis for the carrier material according to the invention.
  • the carrier material according to the invention for carrying out a solid phase assay can, like the sorption material, also be in the form of a shaped body, loose fillings, a sorption material in particle form, in the form of a gel or as a dispersion.
  • the carrier material carries at least one component for carrying out solid phase assays and is also permeable to a fluid.
  • the carrier material has an average pore diameter of 0.1 to 100 ⁇ m, whereby, as with the sorbent material according to the invention, the pore in the case of corresponding materials is understood here as the space between particle-shaped particles and not the porosity of the surface of the corresponding particle.
  • the carrier material has no or only very low non-specific sorption capacity for affinity materials. In contrast, it has a high affinity for certain components which are complementary to the affinity material arranged on the sorption material.
  • these first complementary components are the assay-specific components which, for example, specifically bind an analyte.
  • the components are only poorly unspecifically adsorbable and advantageously bound specifically on or in the carrier material by means of a first, well nonspecifically adsorbable other, complementary component. that will.
  • the carrier material carries at least one easily accessible, inexpensive, stable and / or well-preserved first component which is later, for example when preparing the assay or just before the assay is carried out, a more expensive, unstable, less durable and / or highly specific assay component binds.
  • the carrier material according to the invention can be understood as a quasi at least two-stage carrier material, namely in the first stage as a carrier material with the first, primary non-specifically adsorbed affinity component and further as an at least second stage, in which a second, secondary, complex to the first affinity component ⁇ mental affinity component is bound.
  • the second, secondary affinity component represents the actual affinity component currently to be used in the assay.
  • the carrier material according to the invention is standardized by the proviso that the component located on a specific unit volume of the carrier material for carrying out solid phase assays (affinity material) is its complementary component, with a single homogeneous flow of a specific amount of a specific solution of this complementary component in a specific Concentration of this complementary component, specifically in an amount such that it fluctuates between several statistically relevant loading processes around an average loading value by at most ⁇ 40%.
  • the fluctuation is in particular not more than ⁇ 30%, preferably + . 20% and in particular preferably not more than + .10%.
  • the bound amount remains constant in the mentioned area even in the case of several subsequent flow processes of suitable other liquids on the carrier material. Again, eluting liquids are not suitable.
  • Blocking steps of specific or non-specific adsorption sites in particular washes of the with the Component for carrying out a solid phase assay loaded carrier material, further affinity binding steps for labeling, and / or reinforcement or for binding the actual assay component (s) etc. into consideration.
  • the carrier material according to the invention is advantageous because it can be manufactured relatively easily, without great expenditure of time and equipment, and in particular easily for the user himself, from easily available components, namely, on the one hand, a sorbent, in particular a sorbent according to the invention , as well as the affinity material or materials required for carrying out a solid phase assay as well as a flow-through vessel.
  • buffer solutions can also be offered, which can be used to load the sorption material and carry out the assay.
  • the support material according to the invention can be obtained by loading the sorption material according to the invention in a single flow through a certain amount of at least one solution with a certain concentration of at least one component for carrying out solid phase assays.
  • the carrier material carries at least one component for carrying out the fixed phase assay significantly below the maximum sorption capacity of the sorption material and free unspecific sorption sites of the sorption material are blocked.
  • the carrier material according to the invention can also be converted into another carrier material according to the invention by a specific binding step.
  • the carrier material according to the invention carries in particular an affinity material made of molecules, groups of molecules or particles with affine properties for other substances.
  • the affinity material is particularly selected from the group of enzymes, substrates which interact with enzymes, antibodies, antigens, such as high molecular substances or pollen or other allergens, haptens, biotin or streptavidin, nucleic acids from RNA or DNA Type, in particular those which can be hybridized with other nucleic acids, receptor or ligands of a receptor, viruses, bacteria, cells, cell organelles, blood cells, particles, such as colloidal particles of metals, metal oxides, polymers or combinations of the affinity materials mentioned.
  • the carrier material according to the invention can also be produced from the sorption material according to the invention by bringing the analyte or analytes into contact with the sorption material without prior modification with non-specific material.
  • the analyte or analytes then serve in the next specific adsorption step, e.g. B. a detection step, as specific binding parameters for complementary partners, e.g. B. in detection solutions.
  • the device according to the invention is preferably a hollow body, in the lumen of which one or more of the sorption materials and / or carrier materials according to the invention are arranged. If one or more carrier materials with different components for solid phase assays are arranged in the device according to the invention, this is done in such a way that a homogeneous flow through the hollow body, in particular the areas with carrier material, is ensured. In particular, there should be no edge effects or other areas in which the flow is faster or slower than at other points. This property is essentially determined by the standardization of the carrier material.
  • an exact fit of the sorption or carrier material with the most exact geometric shape possible is important according to the invention.
  • the sorption and / or carrier materials can be fixed in the form of loose beds or gels by means of devices arranged in the lumen of the hollow body. These devices preferably instruct homogeneous flow behavior for a fluid, in particular solutions with analytes, furthermore the devices in particular have only a low own or preferably no unspecific sorption capacity for affinity materials.
  • a method for producing the carrier material according to the invention is based on the fact that the sorption material according to the invention is treated with one or more affinity materials, essentially in solution with a certain concentration in a certain amount of liquid. This can be done on the one hand in a batch process, on the other hand and preferably, however, in a homogeneous flow in a hollow body with an inlet and outlet opening in which the sorption material is arranged.
  • the latter procedure has the advantage that a standardized carrier material with a high, uniform loading in relation to the concentration used is obtained in a very short time and without any outlay on equipment.
  • a comparable loading in the batch process takes a very long time, since initially only outer areas of the sorbent are loaded, in contrast to the flow-through method in which the entire material, ie. H. also the inner surface that quickly comes into contact with the affinity material.
  • the latter procedure enables the carrier material to be produced directly by the user in a simple manner.
  • the corresponding carrier material can also be produced in larger batches and then placed in devices for carrying out a fixed phase assay.
  • free sorption points of the carrier material are blocked with corresponding substances which are inert in the solid phase assay to be carried out. If necessary, the carrier material is washed one or more times.
  • a method for producing a device for carrying out an assay for determining a plurality of analysis parameters, in which at least one sorption material according to the invention is arranged in the lumen of a hollow body is also described and claimed. In this case, a layer arranged in the hollow body has been loaded in the flow. Free unspecific adsorption sites have been blocked, whereupon the next layer is built up in an analogous manner. It is also possible to produce various loaded carrier materials in advance and then to arrange them accordingly in the form of a layer in the assay device.
  • the solid phase assay according to the invention using the sorption material according to the invention and / or the carrier material according to the invention is carried out by treating the sorption material with a component to be used in the solid phase assay, this component adhering to the sorption material and then the solid phase assay in a manner known per se is carried out.
  • the solid phase assay can be carried out on a sorbent material pretreated in a preliminary stage, in particular on a carrier material loaded with an affinity material.
  • Solid phase assays to be carried out with the particularly preferred affinity columns are already described in general form in DE 41 26 436 AI, 42 08 732 AI or DE 195 00 862 AI.
  • the solid phase assay in the sense of the invention is characterized by an optimization of the parameters loading behavior, pore size, flow time and amount of sorbent, possible additional means for flow correction (frits, pressure), amount of affinity material, amount of sample of the analyte solution and properties of the marker, in particular amplification and flow behavior , out.
  • the solid phase assay is standardized independently of temperature due to the sorption or carrier materials used according to the invention.
  • the solid phase assay is preferably carried out as an affinity assay, in particular as an immunoaffinity assay.
  • the sorption material according to the invention enables particularly simple test configurations which allow any user or assay developer to produce or develop their own assays, in particular affinity assays such as immunoaffinity assays. This basically requires only one device for setting up and / or carrying out an affinity assay with the following features:
  • the sample is then optionally treated with an eluent, so that any specifically bound material on the support material according to the invention is detached and / or the marker used is detached, the eluate,
  • a measurement is carried out in the vessel without elution, for which rectangular flow-through cross sections are preferably suitable in order to carry out the measurement through planar vessel walls.
  • a device is therefore also claimed which enables an affinity assay according to the invention to be set up and / or carried out.
  • a particular advantage of the device is that it can make it easier for the user to set up automatable assays.
  • the device has a device for gripping at least one hollow body which is at least partially filled or can be filled with the sorption material according to the invention, in particular in the form of a tube.
  • a device is provided for loading that which is located in the tube or filled in the tube Sorption material with components for carrying out solid phase assays, while maintaining the carrier material described according to the invention.
  • a device can also be provided with at least one hollow body which is filled with the carrier material according to the invention.
  • the device according to the invention has a device for treating the at least one at least partially filled tube with solutions for washing, binding or eluting components of the solid phase assay or the analyte. It can be advantageous to couple the device according to the invention with units for the detection of solid phase components or analytes.
  • the devices of the device are preferably adapted to known microtitration formats. It is then possible in an advantageous manner in biochemical analysis "and diagnostic equipment used that customarily, on the Mikrotitrationsformat parked are to adapt and use.
  • the standardization and the non-specific loading of the sorbent or the specific loading of a carrier material that carries a primary, not yet assay-specific affinity component, directly during the test with the affinity component used for this allow a sample-oriented Processing in the manual or automated test procedure, which is advantageous compared to the conventional test-oriented processing of an assay.
  • the device according to the invention is in particular a small column open at the top and bottom for single use, which contains an activated solid phase matrix for binding proteins.
  • the device according to the invention with the sorption materials and carrier materials according to the invention shows the advantages of an immunoassay (sensitivity, Selectivity) and the advantages of affinity chromatography (high surface and binding capacity, reagent flow) combined in an immunological test system.
  • the activated solid matrix adsorbs the primary immunological reaction partner via hydrophobic interaction when flowing through. After the free binding sites have subsequently been blocked, the test system can be used immediately or stored temporarily, depending on the stability of the immunological reaction partner.
  • the device according to the invention is particularly advantageous because only about 10 minutes are required for the preparation and the preparation and the subsequent test can also be carried out directly one after the other by hand or in an automatic machine.
  • This has the advantage that work can be carried out in a sample-oriented manner, ie devices according to the invention which have not yet been specified can only be correspondingly specified directly before the test or the tests without loss of time.
  • the test system is processed in accordance with the structure of the immunoassay. So sandwich assays and competitive assays can be carried out, as can the previous preparation of the solid phase in the independent flow of the reagents. The actual assay can therefore also be carried out in about 10 minutes.
  • the device according to the invention can be processed in automated workstations in the microtiter plate format and evaluated in particular in the microtiter plate reader at 510 nm, optionally also at 492 nm.
  • the complete sequence consisting of coating, test processing and evaluation no more than 30 minutes are required, rather less, for example in the range from 20 to 25 minutes.
  • the actual test procedure follows, which is set up in sample application, addition of secondary reagents, addition of the labeling substance, optionally washing steps and the elution step. The evaluation follows directly on the elution step.
  • the sorption material according to the invention can, in cooperation with the device according to the invention and an automated device, be used in a particularly advantageous manner for the development of assays.
  • the device according to the invention in which the sorbent material according to the invention is arranged, is treated with a coupling buffer with antigens and / or antibodies for equilibration and specific coating.
  • the coating is carried out with preferably 750 ⁇ l coupling buffer.
  • a sodium bicarbonate buffer is particularly suitable as the coupling buffer.
  • a rectangular cross section of the flow-through vessel in the region of the carrier bed of, for example, 5 mm ⁇ 1.6 mm at a height of 5 mm per carrier bed is advantageous because of the small cross-sectional area, which is suitable for vertical and horizontal flow.
  • the part of the flow-through vessel lying in front of it in the flow-through direction can have a different, in particular larger cross-section in order to allow larger application volumes.
  • the non-specific binding sites that are still free are blocked with immunologically inert protein in buffer. PBS with a proportion of nonionic surfactants has proven to be particularly advantageous for blocking.
  • the assay developer then still has the option of determining the sample dilution for the assay to be developed in simple experiments and creating a calibration curve once.
  • sufficient one-time calibration curve (or the one-off determination of the cut-off range for qualitative yes / no tests) is preferably carried out by carrying out a comparison test in the upper concentration range (clear positive test) for checking and / or compensating Fluctuations between different measuring devices added, in the case of yes / no tests as in example 7 as well through a negative test.
  • sample dilutions in the range from 1: 301 to 1: 1,001 have proven to be advantageous for infection-serological tests. Due to the size of the devices according to the invention, a suitable sample volume is 250 ⁇ l.
  • Suitable secondary antibodies or antigen conjugates are, in particular, biotinylated reagents with a degree of biotinylation of protein: biotin of about 1: 4.
  • biotinylation of different secondary reaction partners allows the use of a uniform marker-streptavidin conjugate.
  • the degree of biotinylation and the concentration of the secondary reagent used should be matched to one another.
  • concentration of the conjugates provided by the applicant is between 3 and 15 ⁇ g / ml.
  • the volume in which the secondary antibody is brought into the device according to the invention is preferably 250 ⁇ l.
  • Abion Red The color conjugate sold under the name Abion Red has proven to be particularly advantageous as a color marker.
  • Abion Red is a dye particle conjugate also developed for use with the devices according to the invention. It is particularly covalently cross-linked and extremely sensitive (approx. 3 orders of magnitude more sensitive than colloidal gold). If the dye conjugate contains streptavidin, Abion Red couples to biotinylated reagents. Abion Red is the subject of the German patent application 19543 556.7.
  • the dye conjugate can be eluted with 300 ⁇ l of ethanol after the reaction in a microtiter plate and then quantitatively measured in conventional photometers at 510 nm or 492 nm.
  • ABION RED is also particularly suitable for measurement in a flow-through vessel without elution, since it allows optical quantitative determination without additional color reaction.
  • the support materials according to the invention have the advantage that binding reactions carried out with them do not show any dependence on incubation times or temperatures (between 15 and 35 ° C.). Therefore, after the expiry of a Processing step follow the next steps immediately. If, however, highly specific reagents show kinetic effects in a range from 0 to 6 min, it is advisable to determine the influence of the reaction kinetics when developing a test.
  • the system already outlined above is available in the form of an automatic machine which can be used both for test development and for test processing, for example in an analytical laboratory.
  • the throughput of the machine is approximately 100 of the devices according to the invention per hour.
  • the advantage of the test system made available according to the invention is that sample-oriented processing can take place and one is largely independent of the test-oriented processing previously carried out in routine processes. It is therefore possible to work through the corresponding tests one after the other for an analysis to be carried out and to record the sample holistically and not, as in conventional analysis, to be bound to receive a holistic test result only when the respective tests involved those involved Machines are processed. Waiting times until a sufficient number of samples for a particular test are reached.
  • the machine can use the following cross-test reagents, namely buffer and system liquids, such as PBS with low surfactant and high surfactant concentration, water, ethanol and coupling buffer.
  • Color marker concentrate, blocking reagent, BSA, anti-human-IgG, anti-human-IgM, anti-human-IgA, preferably biotinylated, and the devices according to the invention in which the sorbent material is arranged uncoated are provided as reagents.
  • the machine itself provides microtiter plates, sample holders with barcode lasers for primary and secondary sample tubes, and microtiter plate photometers the necessary filters and a data processing system with the appropriate software. This machine then, as described above, works through the work steps also mentioned above. Instead of microtiter plates and microtiter plate photometers, the automat contains a corresponding reading device when evaluated in the flow-through vessel.
  • the sorption material according to the invention with the coated carrier material and the corresponding manufacturing method of the carrier material in connection with the automatable device for setting up and carrying out an affinity assay provides a completely new concept in order to give everyone the design without complicated and expensive machine configurations to enable affinity assays.
  • compositions in kit form containing carrier material or sorbent material according to the invention are also claimed according to the invention, cylindrical hollow bodies with inlet and outlet openings, carrier material or sorbent material preferably already being arranged in the hollow bodies, aqueous solutions for loading the sorbent material according to the invention which already contain dissolved affinity material, or separately from the affinity material in storable form and, if appropriate, non-aqueous solutions for loading the sorbent material according to the invention, which either already contain affinity material in solution or contain the affinity material in the composition separately in storable form and, if appropriate, aqueous solutions for carrying out a solid phase assay together with, if appropriate, further auxiliary reagents or devices for Carrying out solid phase assays.
  • a kit in a typical embodiment contains about 200 columns for immunoassay test developments. This also contains washing buffer, blocking reagent, eluent, a highly sensitive particle suspension dye, aids with a microtiter plate, a hanging frame for 96 devices according to the invention in microtiter format, plexiglass frame, waste tray, test-specific reagents for 24 assays including standard serum, as well as other aids, such as descriptions etc.
  • the kits are regularly unopened for a long time, quite up to 2 years, durable.
  • the sorption material or carrier material according to the invention can also be used in the form of the devices according to the invention for the routine checking of samples which may be contaminated with viruses or bacteria.
  • samples which may be contaminated with viruses or bacteria.
  • blood products in blood banks can be examined for HIV, hepatitis, syphilis, etc.
  • Other infectious components such as hepatitis B core antigen and CMV have not been tested to date. Since these are also not harmless to health, it is desirable to also test for these germs. However, this is not done for economic reasons.
  • the objects according to the invention form the basis for tests which, with a high degree of sensitivity, also make it possible to test several donor sera in the pool in the case of blood products.
  • the assays that can be carried out with the objects according to the invention are also able to examine a pool of several sera, for example up to 10 sera, for the presence of a specific antibody with the same sensitivity as a corresponding single test.
  • a so-called sandwich assay is preferably used, it being possible for the sample to be enriched in the flow through the sorption materials according to the invention.
  • the test is suitable for antigens in which a positive finding is relatively rare and also wherever a large number of sera must be examined. This makes it possible to significantly increase the virus safety of blood preparations in blood banks with an economically justifiable effort.
  • An automated blood group determination (automatic Coombs test) can also be carried out according to the invention can be achieved.
  • a blood group determination must be carried out with every blood donation and when a blood donation is administered. This examination is required by law internationally. The existing examination methods are reliable, but due to a centrifugation step in the assay, they are relatively time-consuming and cannot be automated.
  • the sorption materials or carrier materials according to the invention enable rapid and automatic blood group testing, in particular for blood banks and clinics. This variant of the method according to the invention can be expanded to differentiate special antibodies against blood group antigens.
  • the Heliobacter pylori bacterium can also be tested serologically for diagnosis of gastric ulcers accompanying therapy.
  • Round frit made of polyethylene, 5 mm in diameter, 5 mm in height, precise in shape for the cross section of a flow-through vessel, punched out of filter plates which were produced by sintering from polyethylene powder with a narrow particle size distribution by sieving (particle size below 250 ⁇ m); Nominal pore size 50 ⁇ m (bubble point),
  • Pore size distribution with Coulter porometer from 5.492 to 138.6 ⁇ m, average pore size 13.64 ⁇ m, 1.063% of the pore number over 27.5 ⁇ m, 15.45% over 15.92 ⁇ m (Abion GmbH, Irishlich, Germany), - 28448 PC 17EP97 / 00405
  • Coupling buffer 0.1 M sodium bicarbonate / sodium carbonate, pH 9.2;
  • Blocking buffer 0.1 M sodium phosphate, 0.1 M sodium chloride, 1% BSA (bovine serum albumin), 0.005% Tween 20, pH 7.2;
  • Wash and dilution buffers such as blocking buffers without BSA.
  • the frits are inserted individually, essentially unbent and exactly perpendicular to the flow direction, at the upper edge of the conical part of the flow vessels.
  • the loose particles contained in the frits of the manufacturing process, which disrupt the standardization, in particular unbound polyethylene powder particles, abrasion and dust were removed by intensive washing.
  • E ⁇ was washed at least twice with 750 ul ethanol and with 750 ul bidistilled water. If it was necessary to vent the frit, washing was carried out under slight pressure. In order to improve the washing effect and / or the ventilation, the flow direction could also be reversed in individual washing steps. A check after the production of the corresponding frit batches is recommended and is carried out using the uniformity of the Solution received measurement signal in the subsequent affinity assay. Finally, the frits were washed with 750 ⁇ l coupling buffer.
  • the diphtheria toxoid solution was diluted in a coupling buffer to a final concentration of 5 ⁇ g / ml.
  • 750 ⁇ l each of this solution with an absolute amount of 3.75 ⁇ g toxoid were added to each flow-through vessel with the frit inserted.
  • the frits In free flow under gravity (throughput time approx. 6 min), the frits nonspecifically adsorbed essentially uniform amounts of toxoid, which was particularly evident from the subsequently measured uniform binding of the complementary human anti-diphtheria IgG at a constant concentration. No useful signal could be obtained without intensive washing out of the free particles in the frit. Thereafter, any unspecific adsorption sites still present on the frits were each coated with 750 ⁇ l blocking buffer and washed with 750 ⁇ l washing buffer. The vessels produced in this way are called test vessels below.
  • the diphtheria serum is diluted to the concentrations 10, 7.5, 5, 2.5, 0.75, 0.5, 0, 25 and 0.1 mlU / ml with washing buffer.
  • the biotinylated secondary antibody to 6 ⁇ g / ml, the Abion RED conjugate in a ratio of 1:40.
  • 250 ⁇ l of the diluted samples were applied to the test vessels, then 250 ⁇ l of the solution of the biotinylated antibody and 250 ⁇ l of the Abion RED conjugate .
  • elution was carried out with 300 ⁇ l ethanol each.
  • the optical density of the eluate was determined in the ELISA reader at 492 nm against a zero value (ethanol 95% by volume).
  • the ⁇ o determined calibration curve for all further measurements with materials of the same batch is shown in Figure 1.
  • the calibration curve was created with at least 10 measurements per measuring point, with one for each measuring point The coefficient of variation was below 8% (standard deviation / mean x 100).
  • the marking with large dye particles such as Abion RED, with diameters of approx. 100 to 200 nm leads to a non-linear or at most in the lower concentration range even in the capacity range of the text vessel linear relationship between concentration and signal, which can be due to the measurement method for optical density as well as the behavior of large particles in the assay examined here.
  • the example relates to the temperature dependency at 4 ° C to 39 ° C of the determination of the tetanus antibody concentration in serum and thus the calibration curve with Abion RED marker.
  • the temperature-independent calibration and measurement within the measuring range in the capacity range of the test vessels within the measuring accuracy can be seen from FIG. 3.
  • This example relates to the antigen loading dependence of the determination of the tetanus antibody concentration in serum and thus the calibration curve with Abion RED marker or FITC marker.
  • Serum dilution to 7.5, 5.55, 3.75, 1.83, 0.75, 0.375 and 0.183 mlU / ml; Sample application 50 ⁇ l.
  • Figure 6 shows that there is no difference between the two loads when measured with a fluorescence marker. Nevertheless, it is advisable to work with the high loading volume in the test position in order to achieve a better distribution of the binding sites. This distribution is advantageous for accessibility for the large dye marker particles, since a better signal also results when the distribution is better.
  • This example relates to the dependence on the amount of the sample solution using the example of the tetanus and diphtheria antibody concentration in serum.
  • Coupling buffer solution equal absolute amounts of human anti-tetanus IgG in 750 ⁇ l, 250 ⁇ l, 50 ⁇ l and 10 ⁇ l wash buffer solution.
  • the strong dependence of the signal on the amount of the sample can be seen in Figure 7.
  • the measurement with 50 ⁇ l gives the strongest signal and was the most suitable sample quantity for the tests under investigation.
  • the range up to 100 ⁇ l also provides reliable signals, whereas the high signal values at 10 ⁇ l are not preferable due to the large scatter.
  • an assay can also be carried out with the carrier materials according to the invention even in a small amount.
  • This example relates to the pore size dependence of the determination of the diphtheria antibody concentration in serum with Abion RED markers.
  • Example 7 The experiment was carried out as described in Example 5 b). The sample amount was 250 ⁇ l with 3.08, 2 and 1 mlU / ml; Frits with nominal pore sizes of 100, 50, 35 and 3 ⁇ m were used. The dependence of the signal on the pore diameter is shown in Figure 9.
  • Example 7 The sample amount was 250 ⁇ l with 3.08, 2 and 1 mlU / ml; Frits with nominal pore sizes of 100, 50, 35 and 3 ⁇ m were used. The dependence of the signal on the pore diameter is shown in Figure 9. Example 7
  • IgG antibodies against viral antigens influenza A2, type H3N2.
  • This immunoassay is used for the qualitative detection of IgG antibodies in human serum.
  • the antibody sought in human serum forms an immune complex with the antigen immobilized on the solid matrix (influenza virus A2, type H3N2).
  • the anti-human-IgG-biotin conjugate combines with this complex.
  • Abion Red specifically binds to the biotin conjugate via streptavidin.
  • a deep red dye complex is formed (in the case of positive samples), which is eluted with 300 ⁇ l of ethanol in a microtiter plate and measured at 510 nm (possibly 492 nm) in the microtiter plate reader. Unbound reaction partners are removed beforehand by washing processes.
  • the devices according to the invention ethanol absolutely undenatured, coupling buffer, PBS with a high surfactant content, bidistilled water, Abion Red, anti-human-IgG (monoclonal), (influenza A2 virus antigen type H3N2), control serum influenza A2 negative, control serum influenza A2 positive, used.
  • the device according to the invention is suspended in the sample feed of the assay machine and positioned over a washing tub.
  • the devices according to the invention are deaerated by applying 500 .mu.l of ethanol directly with a sharp jet from the multipipette and the amount of liquid passes through the device according to the invention in flow (approx. 2 min). Then 500 ⁇ l, 50 vol .-% ethanol are applied with a multipipette.
  • the influenza A2 virus antigen is adjusted to a protein content of 2.5 ⁇ g / ml in the coupling buffer, and 750 ⁇ l of the solution are adjusted to an amount according to the invention Device abandoned.
  • the throughput time here is typically 4 min.
  • the device according to the invention is blocked with 750 ⁇ l PBS with a high tween content (approx. 3 min) and is now ready to carry out the influenza assay. If necessary, it can also be stored for up to 6 months. Storage should take place at 4 ° C. in a refrigerator and under a PBS buffer which contains, for example, sodium azide as bactericide. If several devices are to be prepared that are not used immediately, the prepared device can also be stored in the sealed bag at 4 ° C for approx. 14 days.
  • the required number of the prepared devices is suspended in a support frame and positioned over a washing tub.
  • the sera to be tested, negative and positive controls, are diluted 1: 101 in PBS with a high Tween content and 250 ⁇ l of each are applied to a device according to the invention.
  • the lead time is typically around 2 minutes. Since incubation times do not bring improved results, these can be dispensed with entirely.
  • the biotinylated, monoclonal secondary antibody (anti-human IgG) is diluted to 15 ⁇ g / ml in the same buffer, and 250 ⁇ l of each are applied to a device according to the invention.
  • the throughput time is approx. 2 min.
  • Abion Red is diluted 1:30 in the same buffer and 250 ⁇ l of each are placed on a device according to the invention.
  • the throughput time here is also about 2 minutes.
  • a washing step is then recommended. This is done with 750 ⁇ l bidest. -Water.
  • the throughput time is about 2 minutes.
  • the carrier frame with the "devices of the invention is a Mikro ⁇ titer plate po ⁇ itioniert.
  • the erfindung ⁇ iliaen devices are then incubated with 300 ul of ethanol in the microtiter plate eluted. Die ⁇ er step lasts for at least about 3.
  • the microtiter plate is al ⁇ bald in MTP Reader at 510 nm, if necessary, measure 492 nm against 300 ⁇ l ethanol as blank sample.
  • the cleavage product C 3 a is formed, which is also referred to as anaphylutoxin.
  • the test described can be used to quantify C 3 a.
  • This in Humanpla ⁇ ma Komplementpeptid C 3 a is bound in the first step from the immobilized on the solid matrix monoclonal antibody.
  • the second biotinylated monoclonal antibody against C 3 a combines with this complex.
  • the dye Abion Red binds specifically to the biotin conjugate via streptavidin.
  • a red dye complex is formed (in the case of positive samples), which is eluted with 300 ⁇ l of ethanol (96%) in a microtiter plate (MTP) and is measured at 510 nm (possibly 492 nm) in the MTP photometer.
  • Light-bound reaction partners are removed by washing processes.
  • the preparation and coating of the erfindung ⁇ nningen Vor ⁇ directions is carried out as described in the previous example with the difference that are used as antibody anti-C 3 a-antibodies.
  • the concentration of the anti-C 3 a antibody is 3 ⁇ g / ml.
  • the blocking is carried out with 750 ⁇ l of a PBS buffer which contains a higher proportion of tween.
  • the procedure is as described in the previous example.
  • the secondary antibody is diluted to 20 ⁇ g / ml in the buffer mentioned.

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Abstract

L'invention concerne un matériau de sorption sous forme de corps moulé, un tel matériau de sorption en vrac sous forme de particules, sous forme d'un gel, d'une membrane ou intégré dans une membrane ou sous forme de dispersion, ledit matériau de sorption étant apte à fixer de façon non spécifique des matériaux à affinité et pouvant être traversé par un fluide. Ledit matériau de sorption se caractérise en ce que: le diamètre moyen de ses pores est de 0,1 à 100 νm; il fixe les matériaux à affinité de façon non spécifique lorsqu'ils le traversent; il est normalisé en ce qu'un volume unitaire donné dudit matériau de sorption, qui peut absorber les matériaux à affinité à fixer, fixe une quantité qui, selon différents processus de charge appropriés du point de vue statistique, oscille autour d'une valeur moyenne de charge d'au moins ±40 %, lorsqu'il est chargé par un seul courant traversant uniforme d'une certaine quantité d'une solution du matériau à affinité concernée, à une concentration définie, et en ce que, lors du blocage subséquent d'emplacements de sorption non spécifiques libres et lors de plusieurs processus appropriés suivants de traversée par un courant du matériau de sorption, tels que des lavages du matériau de sorption chargé avec le matériau à affinité ou l'amenée de matériaux réagissant par affinité, la quantité fixée par le matériau de sorption reste constante dans la plage mentionnée.
PCT/EP1997/000405 1996-01-30 1997-01-30 Materiau support pouvant etre charge par un flux traversant pour dosages en phase solide WO1997028448A1 (fr)

Priority Applications (6)

Application Number Priority Date Filing Date Title
EP97902265A EP0879416A1 (fr) 1996-01-30 1997-01-30 Materiau support pouvant etre charge par un flux traversant pour dosages en phase solide
DE19780059T DE19780059D2 (de) 1996-01-30 1997-01-30 Im Durchfluß beladbares Trägermaterial für Festphasenassays
JP9527299A JP2000507347A (ja) 1996-01-30 1997-01-30 固相アッセイ用の流動充填可能な支持体材料
EA199800671A EA199800671A1 (ru) 1996-01-30 1997-01-30 Наполняемый в потоке материал-переносчик для твердофазных анализов
BR9707232A BR9707232A (pt) 1996-01-30 1997-01-30 Material de suporte carrevável por fluxo para testes em fase sólida
AU15972/97A AU1597297A (en) 1996-01-30 1997-01-30 Carrier material loadable by a through flow for solid phase assays

Applications Claiming Priority (4)

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DE19605003.0 1996-01-30
DE1996105003 DE19605003A1 (de) 1996-01-30 1996-01-30 Im Durchfluß beladbares Trägermaterial für Festphasenassays
DE19615707.2 1996-04-22
DE19615707 1996-04-22

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DE (2) DE19780059D2 (fr)
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Publication number Priority date Publication date Assignee Title
WO1997049991A1 (fr) * 1996-06-25 1997-12-31 Abion Beteiligungs- Und Verwaltungsgesellschaft Mbh Utilisation d'un materiau support pour criblage au moyen de cultures cellulaires

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CN101513606A (zh) * 2004-05-24 2009-08-26 株式会社资生堂 亲和颗粒和亲和分离方法
EP2151688B1 (fr) * 2007-05-30 2018-07-11 JSR Corporation Inhibiteur d'adsorption non spécifique

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WO1987007384A1 (fr) * 1986-05-30 1987-12-03 Quidel Dispositif d'analyse immunologique enzymatique
DE4126436A1 (de) * 1990-09-17 1992-03-19 Abion Ohg Einwegreaktionsgefaess fuer die festphasenimmunanalytik und verfahren zur messung von ueber immunreaktionen bestimmbaren komponenten
DE4115277A1 (de) * 1991-05-10 1992-11-12 Roehm Guenter H Wechseleinrichtung fuer drehfutter-spannbacken
WO1993019368A1 (fr) * 1992-03-18 1993-09-30 Abion Ohg Dr. Erhardt U. Maier Reacteur a usage unique pour l'analyse immunologique en phase solide et procede visant a mesurer les composants dosables par immunoreactions
DE19500862A1 (de) * 1994-01-13 1995-07-20 Abion Ohg Reaktionssäulen für simultane Mehrfachmessung und Verfahren zur Bestimmung von Verbindungen

Patent Citations (7)

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Publication number Priority date Publication date Assignee Title
WO1987007384A1 (fr) * 1986-05-30 1987-12-03 Quidel Dispositif d'analyse immunologique enzymatique
DE4126436A1 (de) * 1990-09-17 1992-03-19 Abion Ohg Einwegreaktionsgefaess fuer die festphasenimmunanalytik und verfahren zur messung von ueber immunreaktionen bestimmbaren komponenten
WO1992005442A1 (fr) * 1990-09-17 1992-04-02 Abion Ohg Reacteur jetable pour analyses immunologiques en phase solide et procede de dosage de composants detectables par reaction immunitaire
DE4115277A1 (de) * 1991-05-10 1992-11-12 Roehm Guenter H Wechseleinrichtung fuer drehfutter-spannbacken
WO1993019368A1 (fr) * 1992-03-18 1993-09-30 Abion Ohg Dr. Erhardt U. Maier Reacteur a usage unique pour l'analyse immunologique en phase solide et procede visant a mesurer les composants dosables par immunoreactions
DE4208732A1 (de) * 1992-03-18 1993-09-30 Abion Ohg Einwegreaktionsgefäß für die Festphasenimmunanalytik und Verfahren zur Messung von über Immunreaktionen bestimmbaren Komponenten
DE19500862A1 (de) * 1994-01-13 1995-07-20 Abion Ohg Reaktionssäulen für simultane Mehrfachmessung und Verfahren zur Bestimmung von Verbindungen

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997049991A1 (fr) * 1996-06-25 1997-12-31 Abion Beteiligungs- Und Verwaltungsgesellschaft Mbh Utilisation d'un materiau support pour criblage au moyen de cultures cellulaires
WO1997049992A1 (fr) * 1996-06-25 1997-12-31 Abion Beteiligungs- Und Verwaltungsgesellschaft Mbh Utilisation d'un materiau support pour la preparation et la detection d'echantillons dans un procede d'analyse en genie genetique

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BR9707232A (pt) 1999-07-20
DE29701511U1 (de) 1998-05-28
JP2000507347A (ja) 2000-06-13
CN1214771A (zh) 1999-04-21
EA199800671A1 (ru) 1999-02-25
EP0879416A1 (fr) 1998-11-25
DE19780059D2 (de) 1999-08-12

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