CN1214771A - Carrier material loadable by through flow for solid phase assays - Google Patents

Abstract

Sorption material in the form of a moulded body, loose bulk sorption material in particle form, in the form of a gel, a membrane or embedded in a membrane or as a dispersion, this sorption material being able to fix affinity materials non specifically and being permeable to a fluid. Said sorption material is characterized in that: it has an average pore diameter of 0.1 to 100 mu m; it fixes the affinity materials non specifically when they pass through is; it is standardised in that a given unit volume of the sorpiton material that is sorptive to the affinity materials to be fixed binds a quantity which, between several statistically-relevant loading processes, oscillates around an average loading value by at most +/-40 % when loaded by a single uniform through flow of a given quantity of a solution of the affinity material at a definite concentration and, that at the subsequent blocking of free non specific sorption positions, and during several following suitable direct through flow processes for the sorption material such as washing of the sorption material loaded with the affinity material or the supply of affinity-reactive materials, the fixed quantity remains constant in the mentioned range.

Description

A flowable carrier material for solid phase analysis

The invention relates to an adsorption material according to the preamble of claim 1, a carrier material for performing a solid-phase analysis according to the preamble of claim 9, a device according to claim 14 , a method of preparing an adsorption material according to the preamble of claim 17, a method of preparing a carrier material according to claim 19, a method of preparing a device for carrying out an analysis according to claim 20 , a kind of solid phase analysis as described in the preamble of claim 21, and a kind of method for loading adsorption material as described in claim 25, a kind of complete apparatus as described in claim 26 and as described in claim 27 a device.

From DE-4126436-A1 and DE-4208732-A1 there are known reaction columns for solid-phase immunoassays and a method for determining components that can be assayed by means of an immunoreaction. These reaction columns are easy to load and unload, can work in continuous flow mode, require only short reaction times, and can perform quantitative reactions. After basic standardization across batches of reaction columns, they require neither pre-calibration or regeneration nor baseline measurements. With this system, known affinity assays such as ELISA plate reactions or immunosorbent reactions can be performed more easily, quickly, precisely and more automatically. However, the relatively lengthy preparation process of such a system still leaves room for improvement due to the relatively large loading of the adsorbent material packed in the reaction column. The loading of the adsorbent material takes a relatively long time. Thus, even gels, such as agarose, can take hours for the loading material to penetrate the micropores. It would be of great advantage to the preparatory work if the loading could be carried out in a defined manner in a shorter time without the use of complex devices and at the same time requiring only a small amount of expensive loading material. Furthermore, it is advantageous if the loading of the adsorbent material for analysis can be done by the user, since, on the one hand, the user has the opportunity to modify the adsorbent material according to the state of the environment. On the other hand, it is also advantageous if the various materials are prepared only a short time before use, so that the demands on their durability are less stringent. Thus, the loaded material required for analysis can be stored separately from the device parts or adsorbent material.

Therefore, the object of the present invention is to overcome the above-mentioned drawbacks and to provide more useful materials. Surprisingly, this object has been achieved by using an adsorption material having the features of claim 1 . After loading with the affinity ligand material, this adsorption material is suitable for use as a support material for solid phase analysis. An advantage of the adsorbent material according to the invention is that it can be loaded rapidly and simply. This sorbent material is standard because it binds to affinity materials with relatively little variability when loaded with them and thus serves as a standardized support for standard solid-phase assays The basis of the material. An advantage of this adsorption material of the present invention is that even with the loading of small amounts of relatively expensive affinity materials, it is still possible to obtain standardized support materials for solid phase analysis. The adsorbent material of the invention can also be considered an intermediate product in the preparation of the support material of the invention. An advantage of this adsorbent material of the present invention is that it can be reproducibly loaded in a continuous flow together with an affinity material for solid phase analysis.

It was especially surprising that the affinity material bound non-specifically to the adsorption material adsorbed very uniformly and reproducibly in a continuous flow, making standardization of this material as well as standardization of the carrier material according to the invention possible.

The adsorption material of the present invention is a shaped body structure, a loosely packed structure of particles, or a gel structure, or as a dispersion. Such an adsorption material of the present invention is capable of binding non-specifically to an affinity material. It must be permeable to a fluid. The average pore size of this adsorbent material is 0.1-100 microns. In the case of packed structures, dispersions and sintered materials, the micropores are the pores present between the particles and not the pores in the surface of the individual particles of the adsorbent material according to the invention. This adsorbent material is capable of non-specifically binding to the affinity material flowing through the adsorbent material. The adsorption material of the present invention is standardized according to the following regulations. When a given amount of this affinity material solution with a given concentration flows through once in a uniform form, the amount of adsorbed affinity material bound to a given unit volume of the adsorption material The quantity varies between several statistically related quantities around an average load value, said variation is not greater than ±40%, especially ±30%, preferably ±20%, most preferably not greater than ±10%.

Possible measurable parameters for controlling the homogeneity of the shaped body are the average weight or density and/or the average flow rate. Changes in shaped body weight and flow rate indicate corresponding changes in parameters such as pore size, total pore volume and internal surface area which, among other parameters, determine the adsorption properties of the support material.

The above-mentioned number ranges remain substantially constant during the subsequent blocking steps of the free adsorption sites and in further processes in which other suitable liquids are present in the adsorption material. Of course, liquids for eluting the adsorbate (eg ethanol, eluting pH solution) are not suitable. In particular, said quantitative ranges are unchanged, for example during the steps of blocking remaining free non-specific adsorption sites, washing of the affinity material-loaded adsorption material and the like.

The variation of the loading value is essentially independent of temperature in the range of 4-40°C. This is an advantage because the loading process can be carried out even in a non-thermostatic laboratory regardless of the ambient state, without further lengthy processing steps, especially in the cold room of a biochemical laboratory.

Preferably, the adsorbent material of the present invention consists of organic or inorganic sintered materials. Organic sintered materials that can be used mainly include thermoplastics, especially thermoplastic particles. These materials include, for example, polyethylene and polystyrene.

In another embodiment, the adsorbent material according to the invention can be a loose packing of particles or a gel structure, a gel-like structure, or a self-supporting shaped body, eg a sinter.

In yet another preferred embodiment, the absorbent material according to the invention consists of foamed plastics, in particular reticular, hollow fibers, oriented sheets, composites or multilayer materials with different properties in terms of adsorption, flow rate or strength, And in order to avoid the formation of large transparent pores, it is composed of ceramic materials, zeolites, metals, metal oxides, alloys, glass, or carbon modified bodies, or combinations of the above materials, in the form of mixtures or multilayer structures.

In general, any homogeneous, porous material that has sufficient non-specific adsorption for the affinity reaction component (affinity material) can be used. These materials include, for example, materials and membranes used in microfiltration, as well as ultrafiltration and deep bed filtration. This adsorbent material can be a sintered shaped body structure produced by sintering under heat and/or pressure, with or without binders and/or macromesh, dense fillers (diatomaceous earth, sand, etc.) stone, anthracite), foams, fibers and hollow fibers, transverse or longitudinal fibers, nonwovens, random fabrics, microfibrils, glass fibers, etc. In addition to the above-mentioned organic materials, other materials and preparation processes can also be used in principle, for example, in particular, in Ullmann Encyclopedia of Industrial Chemistry, Vol.A16, from page 224; 8 ff.; and those described in Siegfried Ripperger, Mikrofiltration mit Membranen, Verlag Chemie, Weinheim 1992, 15 ff.

The homogeneous continuous flow which is relevant to the invention and which is ensured by the inventive adsorption material and carrier material is not only related to a homogeneous pore structure, in particular to the entire surface of the adsorption inner surface, adsorption material or carrier material, but also to its cross-section, i.e. Surface correlation for flow direction. Such a symmetrical uniform pore structure is desirable. However, it is also possible to use asymmetric pore structures, the pores of which, viewed in the direction of flow, are initially small in diameter, for example, to provide a larger internal surface, and later in larger diameters, so that the flow rate does not become too large. slow.

By sintering, it is possible to obtain films with a low porosity (up to 40%, perhaps up to 60%) and a random pore structure with a rather broad pore size distribution. By orienting polymeric multilayer structures, films with porosity up to 90% can be obtained with a relatively regular pore structure and uniform pore size distribution, e.g. from Ullmann, loc.cit., p. 216 arrived. Such films can be produced substantially by methods known in the art.

In a preferred embodiment, the surface of the adsorption material of the present invention may be coated with a biopolymer, such as a protein. Depending on the application field, the surface can also be treated with hydrophobic or hydrophilic treatment.

The process according to the invention for producing the adsorbent material according to the invention begins by sintering a specific screened portion of the sinterable material. Particles in the sintered composition that were not bonded during sintering are then removed from the sintered material.

For standardization, unsuitable particles need to be removed. As a result of such removal, these materials conform to the standardization requirements and cause only small changes in the adsorption amount of the affinity material adsorbed on the adsorption material. This is indicated by a correspondingly smaller change in the measurement signal used to evaluate the solid-phase analysis. This includes measuring the amount of a component that complements the affinity material, such as an analyte component, under otherwise equivalent analytical conditions.

In the case of other adsorbent materials such as foamed plastics, especially foamed plastics of network structure, hollow fibers, unidirectional membranes, composites or sheets, ceramic materials, zeolites, metals, metal oxides, alloys, glass, or carbon In this case, a process step of removing particles of an unsuitable size is also carried out in essence, and/or the process parameters are changed in a known manner to control the pore size, especially the pore size of the membrane material. Achieving a relatively narrow pore size distribution is desirable. Post-treatment may also be required to remove abrasive debris generated during mechanical handling of the material, such as dust generated during shaping of the sorbent material. Such dust has an adverse effect on the adsorption properties and the associated loading. Furthermore, all other impurities, such as solvents and monomers, need to be removed from the adsorption material for standardization in order to be able to achieve standard adsorption.

Reversing the flow direction of the wash liquid in order to remove unsuitable particles, it may be advantageous to use different wash liquids in the individual steps, especially with different hydrophobicity/hydrophilicity, especially for degassing In some cases pressure, vacuum or ultrasound may also be applied.

Desirably, the affinity material is dissolved and preferably allowed to flow in a continuous flow through the loaded sorbent material in a vertical direction and under the force of gravity. In particular, it has proven useful to carry out the loading in a loading container, for example in a cylindrical hollow body. In this case, the adsorption material is arranged in the cavity of the hollow body, that is to say, if possible, it adheres precisely to the wall. This can be achieved on the one hand by producing correspondingly shaped absorbent material shaped bodies and inserting them into the cavity of the hollow body. Another method is to place the adsorbent material between two frits which are preferably non-reactive with the affinity material and with the components that pass through later, especially with components that complement the affinity material. Then, the solution containing the affinity material is injected into the hollow body, which has an inlet and an outlet, so that when the solution containing the affinity material flows through, the latter is non-specifically adsorbed on the surface of the adsorption material.

According to the gravity-dependent vertical flow concept, in the case of a slow flow rate of the adsorbent material, the loading process, which is already carried out at a faster rate, can be achieved by decompression at the outlet of the cylindrical hollow body or pressurization at the inlet end of the cylindrical hollow body. accelerate. This also has a favorable effect on the uniformity of loading. Also, the flow through cylindrical hollow bodies can be decelerated by the use of certain components, such as sinters, or by suction or the application of counter pressure, in order to make the flow of the corresponding material uniform, but mainly to achieve sufficient loading, and Reduce load variation in larger load situations. The same measurement can also be carried out for the subsequent affinity reaction between the affinity component adsorbed on the adsorption material and a complement component flowing through.

The inventive adsorption material serves as the basis for the inventive carrier material. Due to the use of such adsorbent materials, the carrier material according to the invention for solid-phase analysis can also be a shaped body structure, a loosely packed structure of adsorbent material particles, a gel structure, or a dispersion. The support material adsorbs at least one component for solid phase analysis and is permeable to a liquid. Such carrier materials have an average pore diameter of 0.1-100 μm, wherein in the case of the adsorption material according to the invention a pore means the distance between the particles and not the porosity of the surface of such particles in the case of the corresponding material . Such carrier materials have no or only very low non-specific adsorption for the affinity material. Instead, it has a high affinity for certain complement components that bind to the affinity material adsorbed on the adsorbent material.

It is possible to have only these essential complement components as an assay-specific component that specifically binds eg an analyte. In cases where these components are poorly non-specifically adsorbed and are specifically bound to or bound to a carrier material, it may be advantageous to employ a predominant complement component that is readily non-specifically adsorbed. Adsorption of at least one readily available, inexpensive, stable and/or long-lasting principal component to a carrier material, after which, for example, in preparation for analysis or immediately before an analysis is combined with a more expensive, less stable Analytical components that are less persistent, less persistent, and/or highly specific are also highly desirable. In this case, the carrier material according to the invention can be regarded as similar to an at least two-step carrier material, that is to say, a carrier material which acts as the main, essential, non-specifically adsorbed affinity component in the first step, And in at least a second step binds a secondary, auxiliary affinity component that is complementary to the primary affinity component. It is this secondary, auxiliary affinity component that is the appropriate affinity component for practical use in the assay.

The carrier material of the present invention is standardized according to the following provisions, when a given amount of a complement material solution having a given concentration of an affinity material flows through once in a uniform form, the amount contained in a given unit volume of the carrier material is used for solid phase analysis The component (affinity material) will specifically bind to its complement material, and the binding amount varies between several statistically related amounts around an average loading value, and said variation is not more than ±40%. In particular, the variation is not more than ±30%, preferably +20%, most preferably not more than ±10%. This incorporation remains constant within the above-mentioned range of variation, even in subsequent steps in which some of the carrier materials contain other suitable liquids. Likewise, washing liquids are not suitable.

As subsequent steps, in particular, blocking steps of specific or non-specific adsorption sites are possible, in particular washing of the carrier material loaded together with components for solid-phase analysis, as well as labeling and/or amplification, or binding of suitable Other affinity binding steps of the assay components, etc.

The carrier material according to the invention is desirable because it can be prepared in a relatively simple manner without the use of long and complicated devices, especially by the user himself using readily available components, i.e. adsorption materials, especially the present The inventive adsorbent materials and affinity materials or materials required for performing solid-phase analysis, as well as flow conduits are easily prepared. In addition, buffer solutions are available which can be used for loading the sorbent material and for carrying out the analysis process. The support material of the invention can be obtained by adding the adsorption material of the invention to a specific amount of at least one solution having a specific concentration of at least one component for solid-phase analysis. Preferably, the support material adsorbs at least one component for solid-phase analysis significantly below the maximum adsorption capacity of said adsorption material and blocks free non-specific adsorption sites of the adsorption material. A carrier material according to the invention can also be converted into another carrier material according to the invention by specific binding steps.

In particular, the carrier material of the present invention adsorbs affinity materials composed of molecules, molecular clusters or particles that have an affinity for other substances. In particular, this affinity material is selected from the group comprising enzymes, substrates interacting with enzymes, antibodies, antigens, eg high molecular weight substances or pollen or other allergens, haptens, biotin or Streptavidin, nucleic acids of RNA or DNA type, especially those that can hybridize with other nucleic acids, receptors or receptor ligands, viruses, bacteria, cells, organelles, blood cells, particles such as metal colloidal particles, metal Oxides, polymers, or combinations of the aforementioned affinity materials. The support material of the invention can also be prepared from the adsorption material of the invention by contacting the analyte or analytes with the adsorption material without prior modification with a non-specific material. Then, in a subsequent specific adsorption step, eg a detection step, the analyte or analytes are used as a complement component, eg specific binding parameter in the detector solution.

Preferably, the device of the present invention is a hollow body containing one or more adsorbent materials and/or carrier materials of the present invention in its cavity. If one or more carrier materials containing different components for solid-phase analysis are loaded in the device according to the invention, it is carried out in such a way as to ensure a uniform liquid flow through the hollow body, especially in the area. Specifically, there are no edge effects or areas where the flow is faster or slower than elsewhere. This characteristic is essentially determined by standardization of the carrier material. Furthermore, according to the invention, the fit is as precise as possible with a certain geometry (for example, when the flow conduit is cylindrical, a cylinder whose cross-section corresponds to that of the flow conduit, or, when the flow conduit has a rectangular cross-section When cross-section, it is very important that the adsorption material or carrier material is a cube with corresponding cross-section. Adsorbent material and/or carrier material having a loosely packed or gel structure can be immobilized by means of means arranged in the cavity of the hollow body. Desirably, such devices have uniform flow characteristics for liquids, particularly analyte-containing solutions. Furthermore, such devices have only low or preferably no inherent non-specific adsorption for the affinity material.

The method of preparing the support material according to the invention starts from the treatment of the adsorption material of the invention with one or more affinity materials, said affinity materials being substantially in solution, having a specific concentration in a certain amount of liquid. On the one hand, this can be done in batch mode, but on the other hand, it is preferable to do it in a homogeneous continuous flow in a hollow body filled with adsorption material and having one inlet and one outlet. The advantage of the latter method is that standard support materials with high and uniform adsorption rates relative to the concentrations used can be obtained in a very short time and without excessive consumption of equipment. However, it takes a very long time to achieve a comparable adsorption rate using the batch mode, because initially only the outer region of the adsorbent material adsorbs the analyte, as opposed to using the continuous flow mode, where the entire material, i.e. also the inner surface , in rapid contact with the affinity material. Furthermore, the latter method allows the direct preparation of the carrier material by the user in a simple manner. Even if this is not required, corresponding support materials can be prepared in large quantities and then loaded into devices for performing solid-phase analyses.

In a preferred embodiment of the process for preparing the support material according to the invention, the free adsorption sites of the support material are blocked with suitable materials which are inert to the solid-phase analysis being carried out. It is also possible to wash the carrier material one or more times.

The present application also describes or claims a method for producing a device for carrying out an analysis for determining a set of analysis parameters, in which at least one adsorption material according to the invention is contained in the cavity of the hollow body. Each layer arranged in the hollow body is loaded separately in a continuous flow. After blocking the free non-specific adsorption sites, the next layer was prepared in a similar manner. It is also possible to preliminarily prepare differently adsorbed support materials and then arrange them in suitable multilayer structures in the analytical device.

The solid phase analysis of the present invention uses the adsorbent material and/or carrier material of the present invention, and the analysis is carried out by treating the adsorbent material with a component used in solid phase analysis, during the analysis, said component and the adsorbent material binding, followed by solid phase analysis in a well known manner. Alternatively, solid-phase analysis can be carried out on the adsorption material pretreated in the initial step, in particular on the support material to which the affinity material has been adsorbed. In DE-4126436A1, 4208732A1, or DE-19500862A1, descriptions have been given for solid-phase analysis using particularly preferred affinity reaction columns. DE-19500862A1 describes that the advantage of these preferred affinity reaction columns is that the initial reaction of the sample solution can also be carried out in the area above the solid phase analysis area, such as purification, trapping of interfering components or simply extracting the affinity reaction. Analyte response. These advantages can be utilized when performing solid-phase analysis with the adsorption and/or support materials according to the invention.

The solid-phase analysis of the present invention is suitable for quantitative analysis, especially when a predetermined amount of analysis solution is used for analysis, it can perform quantitative analysis on adsorbent materials and carrier materials calibrated within a narrow range. If the volume of the analytical solution remains constant during the solid phase analysis, the volume dependence associated with the solid phase is eliminated.

Solid-phase analysis in the sense of the invention is characterized by the optimization of parameters: adsorption properties, pore size, flow time and amount of adsorbed material, any auxiliary means for flow correction (filter plate, pressure plate), amount of affinity material, analytical The sampling volume of the solution, and the labeling characteristics, especially the amplification characteristics and flow characteristics.

Thanks to the use of the adsorbent or support material according to the invention, solid phase analysis is standardized independent of temperature. Preferably, the solid phase analysis is an affinity assay, especially an immunoaffinity assay.

The adsorbent material of the invention enables an extremely simple assay architecture, which enables each user or assay developer to prepare or develop their own assays, especially affinity assays, such as immunoaffinity assays. In principle, the only thing required is a device for preparing and/or performing an affinity assay, which has the following characteristics: - a device comprising a conduit capable of being stuck and loaded with the adsorbent material of the invention to provide the utensil of the carrier material of the invention placed in the tube; - another or the same utensil for filling the catheter with the carrier material of the invention thus prepared together with the sample; - other binding reactions take place to carry out testing and/or increasing the sensitivity; -then, after washing, the sample can be treated with an eluent, so that the specific binding material on the carrier material of the invention is separated and/or the markers used are dissolved, -then The eluate is collected in a device that allows the measurement of specific parameters of the eluate sample, for example, fluorescence or optical density; - alternatively, measurement is performed without eluate in a container, for which, Rectangular flow duct cross-sections are most desirable to measure with the aid of planar duct walls; - alternatively, to measure specific parameters after a liquid has flowed through.

Therefore, according to the invention, it is also proposed to protect a device capable of preparing and/or carrying out the affinity assays of the invention. A major advantage of this device is that it allows the user to easily prepare for automated analysis. Such a device comprises means for clamping at least one hollow body which is at least partially filled or completely filled with the carrier material according to the invention, the shape of which is particularly preferably tubular. Furthermore, there is provided a device for loading the adsorption material present in the tube or the adsorption material filled into the tube with the solid phase analysis component to obtain the carrier material of the invention as described above. It is also possible to provide a device with at least one hollow body filled with the carrier material according to the invention. In a preferred embodiment, the device of the present invention comprises a means for treating said at least one partially filled tube with a solution for washing, binding or eluting a solid phase analytical component or analyte. It may be advantageous to incorporate devices of the invention with means for detecting solid-phase analytical components or analytes.

Preferably, the individual components of the device of the present invention conform to known microtiter requirements. Thus, it is possible to use this device in biochemical analysis and diagnostics, which are generally suitable for microtiter analysis by definition. The devices according to the invention, in particular devices with several carrier beds, are also suitable as suitable measuring devices (readout devices) for measuring in a fluid conduit. The standardization and non-specific adsorption of the adsorption material or the specific adsorption of the carrier material enables a sample orientation operation during manual or automatic analysis, which is more advantageous compared to the analysis orientation operation in the prior art, wherein The carrier material is capable of adsorbing the main, specific affinity components not yet used for the analysis during the direct analysis with the affinity components.

Specifically, the device of the present invention is a small, disposable reaction column, open at the top and bottom, containing a reactive solid phase matrix for binding proteins. The device of the present invention with adsorbent and carrier materials combines the advantages of immunoassays (sensitivity, selectivity) with the advantages of affinity chromatography (larger surface and binding capacity, flow of reagents) in one immunoassay. analysis system. This reactive solid matrix absorbs the major immune response components through a hydrophilic reaction as they flow through them. Thereafter, after blocking free binding sites, the assay system can be used immediately or stored for later use, depending on the stability of the immune response components. The device of the present invention is desirable primarily because its preparation requires only about 10 minutes, and the preparation and subsequent analysis can be performed immediately in sequence, either manually or automatically. This has the advantage that the operation can be sample-oriented, ie the device according to the invention which has not yet been determined can be determined accordingly immediately before the analysis is carried out without loss of time. The operation of the assay system is designed for immunoassays. As a result, sandwich assays as well as competition assays can be performed, which are also performed in separate reagent streams during the preparatory process for solid-phase assays. Therefore, a suitable analysis can also be performed in about 10 minutes. As mentioned above, the device of the present invention allows for automatic processing and evaluation in a workstation of microtiter plate formats, especially for readings of microtiter plate readers at 510 nm, or at 492 nm. The total run time including coating, assay preparation and evaluation is no more than 30 minutes, or less, approximately 20-25 minutes. A suitable analysis is performed after degassing of the adsorbent material and loading of the degassed adsorbent material, which includes filling of the sample, addition of auxiliary reagents, addition of labeling substances, some optional washing steps, and elution steps. Evaluations were performed immediately after the elution step.

However, in addition to conventional analytical operations, analyzes can also be carried out in a particularly preferred manner with the adsorption material according to the invention and in cooperation with the device according to the invention and an automated instrument. Thus, equilibrating and coating the device comprising the adsorbent material of the invention is treated with a coupling buffer containing antigen and/or antibody. Coating is best done with 750 microliters of coupling buffer. A particularly suitable coupling buffer is sodium bicarbonate buffer. Depending on the type of antigen or antibody, it may be necessary to vary the salt content and pH of the coupling buffer. For the device of the invention, it is sufficient to apply 0.2-10 micrograms of antigen or antibody in the adsorption material of the invention. For low loadings, the cross-section of the fluid conduit in the carrier bed region is preferably a rectangular cross-section because it has a smaller cross-sectional area, e.g. 5 mm x 1.6 mm at a height of 5 mm per carrier bed, which is suitable for vertical flow and horizontal flow. The upstream part of the fluid conduit can have a different, in particular larger, cross-section in order to form a larger filling volume. Blocking of the remaining free non-specific binding sites is performed with immunologically inert proteins in buffer. PBS containing a proportion of nonionic surfactants has proven to be particularly suitable as blocking substance. Therefore, assay developers only need to prepare sample dilutions for developing assays and establish uniform calibration curves in simple experiments. Such a homogeneous calibration curve (or the quantitative cut-off range that uniquely determines whether to proceed with the test) is in principle sufficient, but is advisably supplemented by the use of comparative samples in a higher concentration range (so-called positive samples) to check and/or Or compare the variation between different measurement devices, using negative samples in the yes or no test as in Test Example 7. Thus, in infection immunity assays, sample dilution ratios in the range of 1:301 to 1:1001 have proven to be advantageous. Because of the size of the device of the present invention, a suitable sample volume is 250 microliters. As a secondary antibody or antigen conjugate, there can be used, in particular, a biotin labeling reagent whose degree of biotin labeling is protein:biotin=approximately 1:4. Biotin labeling of different secondary reactants allows the use of uniformly labeled streptavidin conjugates. For some assays, the degree of biotinylation and the concentration of secondary reagents used should be matched. Previous tests have shown that the concentration of the conjugate provided by the applicant is between 3 and 15 micrograms/ml. The volume of the secondary antibody added to the device of the present invention is preferably 250 microliters.

A dye conjugate sold under the trade name "Abion RED" has proven to be particularly suitable as a dye marker. Albion Red is a dye particle conjugate that was developed to also be used in conjunction with the device of the present invention. In particular, it is covalently cross-linked and extremely sensitive (about 3 orders of magnitude more sensitive than colloidal gold). If streptavidin is included in the dye conjugate, Abion Red couples to the biotin labeling reagent. Abien red is the subject of the invention of German patent application 19543556.7. After the reaction is complete, the dye conjugate can be eluted with 300 microliters of ethanol onto the microtiter plate, and then quantitatively measured using a conventional 510 nm or 490 nm photometer. Albion red is also particularly suitable for measurements in fluid conduits without elution, as it allows optical quantitative measurements without secondary color reactions. An advantage of the carrier material according to the invention is that the binding reaction with this carrier material appears to be independent of incubation time or temperature (between 15 and 35° C.). So, after one processing step is completed, the next step can proceed immediately. However, if a specific reagent indicates a kinetic effect between 0 and 6 minutes, it is advisable to determine the effect on the kinetics of the reaction when developing the assay.

For analysis using the device according to the invention, the above-mentioned system is embodied in the form of an automatic instrument which can be used, for example, in an analytical laboratory for conducting and operating the analysis. The throughput of this instrument is about 100 devices of the invention per hour. An advantage of the analysis system provided by the present invention is that the analysis operation can be sample-oriented substantially independently of the analysis-orientation operation performed hitherto in conventional analysis procedures. It is thus possible to carry out a corresponding analysis successfully for an analyte and to process the sample in a comprehensive manner and not, as in routine analysis, be limited only after the various analyzes have been carried out in the corresponding automatic instrument Get a comprehensive analysis result. Therefore, the waiting time for a sufficient number of analytical samples to arrive is eliminated.

This automated instrument can perform different analyzes using reagents, namely buffers and system fluids such as PBS containing low or high concentrations of surfactants, water, ethanol and coupling buffers. As reagents, dye marker concentrates, blocking reagents, BSA, anti-human immunoglobulin G, anti-human immunoglobulin M, anti-human immunoglobulin A, preferably biotin-labeled, and the device of the present invention are provided , which contains adsorbent material without an overlay. The automatic instrument itself is equipped with microtiter plates, sample holders with barcode lasers for the main and auxiliary sample tubes, a microtiter plate photometer with each required filter, and data processing using the corresponding software equipment. Thus, such automated instruments carry out the above-described processing steps in the above-described manner. When the evaluation is carried out in the fluid line, the automatic instrument includes a corresponding reader, which replaces the microtiter plate and the microtiter plate photometer.

Thus, the adsorption material of the present invention as well as the carrier material with the coating and the various preparation methods of the carrier material in combination with an automatic instrument for preparing and performing the affinity analysis provide a completely novel concept, enabling anyone without Affinity assays are designed using complex and expensive device configurations.

According to the invention, a kit is also claimed which comprises the carrier material or adsorbent material according to the invention, a cylindrical hollow body with an inlet and an outlet, preferably already loaded with the Said carrier material or adsorption material, the aqueous solution used for adsorption of the adsorption material of the present invention, said adsorption material already contains the affinity material dissolved therein, or is separated from the affinity material in stored form, and is used for adsorption of the adsorption material of the present invention. The optional non-aqueous solution of the material, said adsorption material either already contains the affinity material dissolved therein, or the affinity material is contained in the kit separately in the form of storage, and optionally for carrying out the solid phase analysis Aqueous solutions, which are separated from other ancillary reagents and devices used to perform solid-phase analysis.

In a typical embodiment a kit contains approximately 200 reaction columns for developing immunoassays. It also includes wash buffers, blocking reagents, eluents, highly sensitive particle suspension colloidal dyes, auxiliaries with microtiter plates, hanging brackets for the device of the invention in 96 microtiter formats, plexiglass holders, Waste tank, analysis-specific reagents for 24 types of analysis, including standard sera, and other accessories such as instructions for use, etc. Typically, in an unopened state, this kit can be stored for a period of time, up to 2 years.

The inventive adsorption material or carrier material can also be used in the form of the inventive device for the routine examination of samples which may contain viruses or bacteria. Thus, for example, blood products in blood banks can be checked for HIV, hepatitis virus, syphilis, etc. Other infectious agents such as hepatitis B core antigen and CMV have not been examined to date. Since the latter are not without health hazards, these germs also need to be analyzed. However, due to economic reasons, this work has not yet been carried out. However, the inventive subject-matter of the present invention forms the basis of a test that enables very sensitive testing of pooled sera from multiple donors in the case of blood products. The assays enabled according to the inventive subject matter can also examine multiple sera, for example a mixture of 10 sera, for the presence or absence of a particular antibody with the same sensitivity as the corresponding individual tests. It is advisable to use a so-called sandwich analysis, in which the enrichment of the sample in a continuous flow can be achieved with the adsorbent material according to the invention. This assay is suitable for examining antigens where positive results are expected to occur infrequently and in cases where large quantities of sera need to be examined. This can greatly improve the safety of blood antiviral preparation in blood banks, and the cost is reasonable. An automatic determination of blood type (automatic Coombs test) can also be realized according to the invention. The blood type must be determined with every blood donation and with the use of the donated blood. Such inspections are mandated by law internationally. The present inspection method, although reliable, takes considerable time due to the centrifugation step used in the analysis and is not automatic. The inventive adsorbent or carrier material enables rapid and automated serum testing, especially in blood banks and hospitals. Variations of this method of the present invention can be extended to the differentiation of specific antibodies and blood group antigens.

In addition, serum tests for bacteria such as Heliobacter pylori can be performed in the therapeutic diagnosis of gastric ulcers.

The present invention is further illustrated below by test examples. Test example 1

Determination of Diphtheria Antibody Levels in Serum and Establishment of Uniform Calibration Curve Materials

Circular polyethylene sinters, 5 mm in diameter and 5 mm in height, precisely fitted to the cross-section of the fluid conduit, made by sintering polyethylene powder with narrow particle diameter distribution obtained by screening (particle diameter less than 250 microns) The filter plate is stamped and obtained; the nominal pore size is 50 microns (bubble point), the pore size distribution measured with a Coulter porosity meter is from 5.492 to 138.6 microns, the average pore size is 13.64 microns, and 1.063% of the voids are greater than 27.5 microns; 15.45% of the voids are greater than 15.92 Micron (Abion GmbH, Julich, Germany);

As described in DE-4126436A1, there is a circular, hollow fluid conduit with a conical lower part, 5 mm in diameter, with a sample solution capacity of about 0.8 ml through a 5 mm high frit fixed at the upper end of the conical shape (Abion Limited company, Julich, Germany);

SVM's Diphtheria Toxoid, NL-3723 BG Bilthoven, The Netherlands;

Biotin-labeled anti-human immunoglobulin G antibody (Sigma, Munich, No.B-1140);

Abion red/streptavidin conjugate, Abion Ltd, Julich;

Diphtheria-positive serum with a content of 1.7IU/ml (IU is the international unit) and diphtheria-negative serum (Abion Co., Ltd., Julich);

Ethanol, the absolute content is 95% (volume);

Coupling buffer: 0.1M sodium bicarbonate/sodium carbonate, pH 9.2;

Blocking buffer: 0.1M sodium phosphate, 0.1M sodium chlorate, 1% BSA (bovine serum albumin), 0.005% Tween 20, pH 7.2;

Wash and Elution Buffer: Same as Blocking Buffer, but without BSA. process

At the upper edge of the conical part of the fluid conduit, the sinter is inserted into the fluid conduit, respectively, substantially straight and exactly perpendicular to the direction of flow. Loose particles contained in the sinter produced during its preparation, especially unbound polyethylene powder particles, abrasives and other dust, which interfere with the calibration, are removed by thorough washing. Wash at least twice with 750 µl of ethanol and 750 µl of double distilled water. If it is necessary to degas the sinter, the washing is carried out at a lower pressure. Furthermore, in order to increase the washing effect and/or the degassing effect, the flow direction can be reversed in some washing steps. It is advisable to check after each batch of sinter has been prepared by checking the homogeneity of the measurement signal obtained with the sample solution in the subsequent affinity analysis. Finally, the frits were washed with 750 μl of coupling buffer, respectively. antigen coupling

Diphtheria toxoid solution was diluted with coupling buffer to a final concentration of 5 μg/ml. 750 microliters of this solution, containing an absolute amount of 3.75 micrograms of toxoid, was injected into each fluidic conduit with an inserted frit. In free gravity flow (flow time approximately 6 minutes), the sinter non-specifically adsorbs a substantially uniform amount of toxoid, which can be obtained from, inter alia, the uniform incorporation of a fixed concentration of complement human anti-diphtheria immunoglobulin G (measured later). obtained) can be seen. If the free particles in the sinter are not thoroughly cleaned, no useful signal can be obtained. Afterwards, the remaining nonspecific adsorption sites of the frits were filled with 750 μl of blocking buffer, followed by washing with 750 μl of washing buffer. The fluid conduit thus prepared is hereinafter referred to as a test vessel.

Diphtheria sera were diluted with wash buffer to concentrations of 10, 7.5, 5, 2.5, 0.75, 0.5, 0.25 and 0.1 mIU/ml. The biotin-labeled secondary antibody was diluted to 6 μg/ml, and the Albion red conjugate was diluted 1:40. 250 microliters of each diluted sample was added to the test vessel, followed by 250 microliters of biotin-labeled antibody solution, and 250 microliters of Albion red conjugate. After washing with 750 microliters of washing buffer, elution was performed with 300 microliters of ethanol, respectively. The optical density of the eluates was determined relative to a reference (95% by volume ethanol) on a 492 nm ELISA reader. In Figure 1 a calibration curve established from all other measurements obtained using the same batch of material is represented. This calibration curve was constructed using at least 10 measurements per measurement point such that the coefficient of variation per measurement point was less than 8% (standard deviation/mean x 100).

In contrast to the results obtained with small fluorescent labeling molecules, labeling with large dye particles such as Albion Red (which is about 100 to 200 nm in diameter) gives a concentration-signal relationship even in testware It is not linear in the capacity range, or only linear in the lower concentration range. This can be explained by the densitometry method and the nature of the large particles detected in the analysis. Test example 2

Comparison between measured values of tetanus antibody concentrations in serum and calibration curves using fluorescent and dye markers. Material:

Same as Test Example 1, except the following parts: Tetanus toxoid of SVM, NL-3723 BG Bilthoven, Netherlands; Tetanus-positive serum (IU is international unit) with a concentration of 14.1IU/ml; and Tetanus-negative serum (Abion Ltd, Julich); 1:20 dilution of AbionRED conjugate; FITC streptavidin conjugate from Dianova (Hamburg); coupling buffer at pH 9.5 Procedure:

Same as described in Test Example 1, except for the following: Tetanus serum was diluted to concentrations of 10, 7, 4, 2.44, 2, 1.51, 1, 0.74, 0.5, 0.244, and 0.1 mIU/ml with washing buffer, And dilute the FITC conjugate solution according to the ratio of 1:251. Add sample: 750 µl of sample solution. From Figure 2 it can be seen that the relationship between signal and concentration using fluorescent markers is linear over the volume of the test vessel, as opposed to the curvilinear relationship established in the case of dye markers. Test example 3

This test example concerns the temperature dependence of tetanus antibody serum assays at 4-39°C and a calibration curve obtained using the Albion red marker. The test was essentially the same as Test Example 2, except that fewer serum dilutions were used; all materials were used immediately at 4°C, 18°C or 39°C. It can be seen from Fig. 3 that both the calibration curve and the measured value are independent of temperature within the measurement accuracy range within the temperature range of the capacity range of the test vessel. Test example 4

This test example involves the correlation of tetanus antibody serum assays with antibody adsorption and calibration curves obtained using either Albion red or FITC markers.

a) Higher adsorption concentration range, the labeling process of Abien Red is the same as that of Test Example 2, the difference is that less serum diluent is used; the dilution ratio of Abien Red conjugate is 1:20, and the antibody-conjugated The concentration was 5 μg/ml and the toxoid concentration was 10 μg/ml (3.75 μg and 7.5 μg in absolute amounts, respectively). It can be seen from Fig. 4 that the signal is independent of the adsorption amount in this adsorption capacity range.

b) Lower range of FITC-labeled adsorption concentration process is the same as treatment 2; the antibody coupling concentration is 5 μg/ml, 2.5 μg/ml, 1.25 μg/ml (absolute amount is 3.75 μg, 1.88 μg and 0.94 μg). Serum dilutions were 7.5, 5.55, 3.75, 1.83, 0.75, 0.375, and 0.183 mIU/ml; add 50 microliters of sample. From Fig. 5 it can be seen that the signal is related to the adsorption amount in the range of adsorption capacity.

c) The amounts added to the adsorption solution have equal absolute amounts. The test was carried out in the same manner as described in Test Example 4b); the amount of antibody coupling was 750 µl at a concentration of 2.5 µg/ml and 250 µl at a concentration of 7.5 µg/ml. Figure 6 shows that there is no difference between the two adsorption amounts when measured with a fluorescent marker. In any case, it is recommended to operate at high adsorption capacity in preparation for analysis in order to achieve a better distribution of binding sites. This distribution is advantageous for the accessibility of labeling particles with large dyes, as a better distribution also results in a better signal. Test example 5

This test example concerns the correlation of serum levels of tetanus and diphtheria antibodies with the volume of sample solutions.

a) Use FITC markers: the test is carried out in the same manner as Test Example 4; 3.75 micrograms (absolute amount) of tetanus toxoid are added to the coupling buffer solution of 750 microliters; Human anti-tetanus immunoglobulin G was added in equal absolute amounts to microliter and 10 microliter wash buffer solutions. From Figure 7 it can be seen that the signal has a strong dependence on the sample size. Within this range, the measurements performed with 50 microliters of the solution yielded the strongest signal with a magnitude in the most suitable analytical signal range obtained with respect to signal optimization. Extending this range to 100 µl still yields reliable signal, whereas the high signal values obtained at 10 µl are undesirable due to the higher diffusivity. However, analyzes can also be carried out at lower amounts using the carrier material according to the invention.

b) The Abien red marker process is the same as described in Test Example 1; sinter: 3.2 mm high, nominal pore diameter 3 microns, the pore diameter range measured with a Coulter porometer is 3.156 to 80.33 microns, and the average pore diameter is 6.046 microns, 1.304% of the pores were larger than 11.25 microns, serum samples contained 0.77, 0.5 and 0.25 μIU (absolute amount). The strong dependence of the signal on the sample size in this case can be seen from FIG. 8 . When using this marker, 750 microliters was found to be an appropriate sample volume for this marker. Test example 6

This test example concerns the correlation of diphtheria antibody serum level measurements with pore size using the Albion red marker. This test was carried out as described in Test Example 5b). The sample volume was 250 microliters at concentrations of 3.08, 2 and 1 mIU/ml; frits with nominal pore sizes of 100, 50, 35 and 3 microns were used. From Figure 9 it can be seen that the signal is dependent on the aperture. Test Example 7

Detection of Immunoglobulin G Antibody Against Viral Antigen (H3N2 Influenza Virus A2)

This immunoassay is used for the quantitative detection of IgG antibodies in human serum. The antibody found in human serum is an immune complex formed by binding to an antigen (H3N2 influenza virus A2) immobilized on a solid substrate. Anti-human immunoglobulin G/biotin conjugate binds to this complex.

Albion red is conjugated to biotin via streptavidin. This (with positive samples) forms a magenta dye complex which is diluted into microtiter plates with 300 microliters of ethanol and measured on a microtiter plate reader at 510 nm (492 nm is also possible). Unbound reaction components are initially removed by washing methods. In this test example, the device of the present invention, ethanol (absolutely pure, non-denaturing), coupling buffer, PBS with high content of surfactant, double distilled water, Albion red, anti-human immunoglobulin G (monoclonal), (H3N2 influenza virus antigen), control serum influenza virus A2 negative and control serum influenza virus A2 positive.

The device of the present invention is suspended in the sample source of an automated analyzer and positioned above a sink. The device of the invention was degassed by injecting 500 microliters of ethanol from a multi-tip pipette directly into the pointed nozzle and passing this amount of liquid through the device in a continuous stream (approximately 2 minutes). Then, 500 microliters of 50% strength (volume) ethanol was injected with a multi-tip pipette.

Using coupling buffer, the protein content concentration of influenza A2 virus antigen was adjusted to 2.5 μg/ml, and 750 μl of the solution was added to the device of the present invention. The flow time is typically 4 minutes. After covering, the device of the invention was blocked (approximately 3 minutes) with 750 microliters of PBS with a high Tween content and the influenza virus analysis was now ready. In addition, it can be stored for up to 6 months. Storage should be kept in the refrigerator at 4°C and PBS buffer containing eg sodium azide as bactericide is used. If multiple devices are prepared and not used immediately, the prepared devices can be stored in a sealed plastic bag at 4°C for approximately 14 days.

For analysis, the required number of prepared devices is suspended in a carrier and set above the sink. Serum to be tested, negative and positive control agents were diluted 1:101 with PBS with high Tween content, and 250 microliters of each solution was added to the device of the present invention. The flow time is typically 2 minutes. Since the incubation time does not improve the results in any way, they can be completely ignored. Biotin-labeled monoclonal secondary antibody (anti-human immunoglobulin G) was diluted to 15 µg/ml with the same buffer, and 250 µl of each solution was added to a device of the present invention. The flow time is about 2 minutes. Albion Red was then diluted 1:30 with the same buffer and 250 microliters of each solution was added to a device of the present invention. The flow time in this case is also about 2 minutes. It is advisable to perform a washing step with 750 microliters of double distilled water. The flow time is also about 2 minutes in this case. Afterwards, the rack containing the device of the invention was placed over a microtiter plate. The device of the invention was then eluted with 300 microliters of ethanol in a microtiter plate. This step takes about 3 minutes. Then, immediately use a 510nm (or 492nm) MTP reader to measure with 300 microliters of ethanol as a reference solution. Test example 8

Sandwich immunoassay for quantitative determination of complement peptide _C3a .

During the activation of the complement system, mainly the cleavage product C ₃ a is formed, which is also called anaphylatoxin. Using the assay, _C3a can be quantitatively determined. In the first step, complement peptide _C3a in human plasma is bound to a monoclonal antibody attached to a solid substrate. A secondary biotinylated monoclonal _C3a antibody binds to this complex.

The dye Albion red is specifically bound to a biotin conjugate via streptavidin. A red dye complex is formed (using a positive sample), which is eluted with 300 µl of ethanol (96%) into a microtiter plate (MTP) and measured using a 510 nm (492 nm) MTP photometer Measurement. Unbound reactants are removed by washing steps. The device of the present invention was prepared and covered as described in the foregoing test example, except that an anti _-C3a antibody was used as the antibody. The concentration of anti _-C3a antibody was 3 μg/mm. The blocking step was performed with 750 microliters of PBS buffer with a higher Tween ratio. The procedure was the same as described in the previous test example. This secondary antibody was diluted to 20 μg/ml with the buffer described.

Claims (28)

1, a kind of sorbing material, it has the loose packed structures, gel structure, film of formed body structure, absorbing material granules or embeds in the film or as a kind of structure of dispersion, wherein said sorbing material can combine with affinitive material non-specificly, and for fluid is permeable, it is characterized in that:

The average pore size of said sorbing material is the 0.1-100 micron;

Said sorbing material combines with the said affinitive material that flows through sorbing material non-specificly;

Said sorbing material carries out standardization according to following provisions, when the solution of the said affinitive material with given concentration of specified rate flows through one time equably, the given unit volume of loading sorbing material combines with the affinitive material that is adsorbed, binding capacity is changing between a plurality of statistical dependence charge capacity of an average load amount, and variable quantity is no more than ± and 40%; With

In the step and a plurality of suitable flowing process step of the free non-specific adsorption point of follow-up sealing to sorbing material, for example in the supplying step to the washing step of the sorbing material that is loaded with affinitive material or compatible reaction material, above-mentioned binding capacity upper lower limit value remains unchanged in above-mentioned scope.

2, sorbing material as claimed in claim 1 is characterized in that the variation of said charge capacity is temperature independent basically in 4-40 ℃ of scope.

3,, it is characterized in that said sorbing material has hydrophobic substantially and/or water-wetted surface as claim 1 and/or 2 described sorbing materials.

4,, it is characterized in that said sorbing material is made of the organic or inorganic material of sintering as the described sorbing material of claim 1 to 3.

5, as at least one described sorbing material in the claim 1 to 4, it is characterized in that said agglomerated material by the thermoplastic particle, for example tygon or polystyrene particle constitute.

6, as at least one described sorbing material in the claim 1 to 5, it is characterized in that said sorbing material be loose particles packed structures or gel structure, gel-like structure, as film or embed in the film, perhaps as a kind of self-supporting formed body, for example a kind of clinkering thing.

7, as at least a described sorbing material in the claim 1 to 3, it is characterized in that said sorbing material is by polyfoam, the composition of particularly cancellated polyfoam, hollow fiber, orientation sheet material, compound substance or multilayer material, stupalith, zeolite, metal, metal oxide, alloy, glass or carbon or above-mentioned material, perhaps sandwich construction constitutes.

8, as at least one described sorbing material in the claim 1 to 7, it is characterized in that said sorbing material can be by using biopolymer, as protein, the surface of coating sorbing material changes the hydrophobicity on surface and/or water wettability and obtains.

9, be used to carry out a kind of carrier material of solid phase assays, it has loose packed structures, gel structure, the film of formed body structure, absorbing material granules or embeds film, perhaps as a kind of dispersion, wherein said carrier material has at least a composition that is used to carry out solid phase assays, and for a kind of fluid is permeable, it is characterized in that:

The average pore size of said carrier material is the 0.1-100 micron;

Said carrier material does not have or has only low-down non-special adsorbability for affinitive material;

Said carrier material carries out standardization according to following provisions, when the complement component dilute solution of the complement component of the composition that is used to carry out solid phase assays (affinitive material) with given concentration of specified rate flows through one time equably, the composition that is used to carry out solid phase assays (affinitive material) that comprises in the carrier material of given unit volume combines specifically with its complement component, binding capacity is changing between a plurality of statistical dependence charge capacity of an average load amount, and variable quantity is no more than ± and 40%; With

Even handle in some steps of carrier material at the liquid flow that follow-up usefulness is fit to, for example be loaded with the step of carrier material of the composition that is used for carrying out solid phase assays and the step of supply compatible reaction material in step, the washing of sealing specificity or non-specific adsorption point, the charge capacity upper lower limit value also remains unchanged in above-mentioned scope.

10, carrier material as claimed in claim 9, it is characterized in that said carrier material can by load in the continuous stream of at least a solution of at least a composition that is used for carrying out solid phase assays that comprises limiting concentration of limited amount as in the claim 1 to 8 the described sorbing material of at least one item obtain, wherein saidly at least aly be used to carry out the composition of solid phase assays or directly combine with sorbing material in a kind of non-specific mode, perhaps in a kind of non-specific or specificity mode, combine indirectly with at least a composition on being combined in said sorbing material.

11,, it is characterized in that said carrier material brings to less a kind of composition that is used to carry out solid phase assays below the maximum adsorption capacity of sorbing material, and the free adsorption site of sorbing material seals as claim 8 and/or 9 described carrier materials.

12,, it is characterized in that said affinitive material constitutes by molecule, molecular radical or with particle that other material has affinity as at least one described carrier material in the claim 9 to 11.

13, carrier material as claimed in claim 12, it is characterized in that said affinitive material is selected from the group that comprises following material: enzyme, with substrate, antibody, antigen, for example polymer substance or pollen, haptens, biotin or strepto-microbiotic, RNA or the DNA type nucleic acid of enzyme interacting, particularly those can with ligand, virus, bacterium, cell, organelle, haemocyte, particle other nucleic acid hybridization, acceptor or acceptor, for example composition of metal colloidal particle, metal oxide, polymkeric substance or above-mentioned affinitive material.

14, a kind of device that has a ducted body, in its cavity to guarantee that the mode that evenly flows is mounted with as any described one or more sorbing materials in the claim 1 to 8 and/or as any described at least a carrier material in the claim 9 to 13 and/or be loaded with as being used for the variety carrier material of the heterogeneity of solid phase assays among the claim 9-13 as described in each.

15, device as claimed in claim 14 is characterized in that the utensil that said sorbing material and/or carrier material utilization are arranged in the cavity of said ducted body is fixed with the form of loose accumulation, film, sheet material or gel structure.

16, as claim 14 and/or 15 described devices, it is characterized in that described utensil set-up mode guarantees evenly to flow and/or specific flow rates, and do not have or only have lower intrinsic non-special adsorbability for the said affinitive material that is adsorbed on said sorbing material or the carrier material.

17, be used for preparation method, it is characterized in that in order to carry out standardization as any described sorbing material of claim 1 to 8:

But under the situation of agglomerated material, the specific sieving part of the said material of sintering; With

Under the situation of other sorbing material, the gap structure of controlling said material is to reach narrower pore diameter distribution, a kind of aftertreatment of the line option of going forward side by side; And under above each situation

From sorbing material, remove unaccommodated discrete particles and material, particularly also carry out forming step thereafter.

18, method as claimed in claim 17 is characterized in that for providing the said standardization that sorbing material or carrier material carry out original material be to carry out in having a device of a ducted body.

19, be used for preparing a kind of method as any described carrier material of claim 9 to 13, it is characterized in that handling as any described sorbing material in the claim 1 to 8 with one or more affinitive materials, various affinitive materials all exist in solution with specific concentrations in specified quantitative liquid, particularly in sorbing material being housed and having equal uniform flow in the ducted body of an inlet and an outlet, be the step of free adsorption site of sealing and optionally washing subsequently, the wherein said affinitive material that is adsorbed on the sorbing material also can randomly combine with the special-purpose affinitive material of analysis.

20, a kind of method that is used to a kind of device of preparing to analyze, said analysis is used for determining a group analysis parameter, wherein will be contained in the ducted body as at least one described at least a sorbing material in the claim 1 to 8, each layer material that it is characterized in that being arranged in the ducted body loads respectively in continuous stream, and the sealing ionization point prepares layer of material down thereafter in the same way.

21, use as at least one described sorbing material of item in the claim 1 to 8 with as at least one a kind of solid phase assays that described carrier material carries out in the claim 9 to 13, it is characterized in that being used for the said sorbing material of a kind of one-tenth divisional processing of solid phase assays, said composition is attached on the said sorbing material, carry out solid phase assays then, perhaps utilize sorbing material pretreated in an initial step to carry out said solid phase assays.

22, solid phase assays as claimed in claim 21 is characterized in that carrying out quantitative test by demarcating (standardized) when using quantitative test solution.

23, solid phase assays as claimed in claim 22 is characterized in that carrying out temperature independent demarcation.

24, as at least one described solid phase assays in the claim 21 to 23, it is characterized in that said analysis is designed to affinity analyzing particularly immune affinity analyzing.

25, be used for loading sorbing material and/or be used in the method for carrying out compatible reaction as any described solid phase assays of claim 21 to 24, it is characterized in that by reducing or increase some the component affects fluids in pressure or the use ducted body, for example flow rate of the sinter of Shi Heing.

26, a kind of complete sets of instrument, it comprises as the described carrier material of claim 9 to 13 or as the described sorbing material of claim 1 to 8, a plurality of cylinder bodily form ducted bodies with entrance and exit, wherein said carrier material or sorbing material preferably have been arranged in the said ducted body, the aqueous solution that is used for load sorbing material of the present invention, said sorbing material has comprised the affinitive material that is dissolved in wherein, but perhaps placed apart with storing mode and affinitive material, with other the optional solution that is used to carry out solid phase assays, and other is used for the optional auxiliary reagent or the device of solid phase assays.

27, be used for preparing and/or carry out a kind of device as at least one described a kind of affinity analyzing of claim 21 to 24, it comprises and is used for blocking with at least one ducted body partially filled at least as at least one described sorbing material of claim 1 to 8 or that fill, test tube particularly, load on the sorbing material of said test tube with the composition that is used for to carry out solid phase assays, to obtain a utensil, perhaps comprise the utensil that at least one uses the ducted body that described carrier material filling maybe can be filled as at least one item in the claim 9 to 13 as at least one described a kind of carrier material in the claim 9 to 13.

28, device as claimed in claim 27, it is characterized in that comprising be used to wash, in conjunction with and/or the solution of wash-out solid phase assays composition or analyte inject utensil of said at least one partially filled at least test tube and optional at a kind of liquid that has flow through and/or at the detecting device of the analyte of eluent check and analysis composition or ducted body.

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