DE2806389A1 - Multiple, automated immunoassay method - with reaction carried out in removable open=ended syringe tips - Google Patents

Abstract

Determination of the amt. of analyte (A) in a sample, where (A) is one of a pair of specific bindable substances, and one member of the pair is insolubilised, uses as detectant (D) one member of the pair giving a measureable signal proportion to the number of molecules of (D) present. The improvement is that first a soln. or (A) is drawn, by syringe, into an open-ended tip, detachably mounted on the syringe, which contains the insolubilised component. (where (A) can be bound to the tip it may be the insolubilised component). A soln. of (D), and other reagents or wash solns. is supplied to the tip (from an outside source or from the syringe), then the solns. removed from the tip by the syringe. The amt. of (D) insolubilised in, or remove from, the tip is then measured to determine the amt. of (A). Process is esp. for immunoassay procedures such as radioimmunoassay (RIA), immunoradiometric assay (IRMA) or 2-site IRMA. The system requires minimal supervision and avoids fluctuating conditions.

Description

description

The invention relates to a device and a method for multiple analysis.

The use of immunoassays (immunobiological detection methods) for the detection of compounds of physiological interest is increasingly gaining ground Meaning since disease states have been found to be related to the Presence or absence of compounds or the concentration of a specific one Link. Because of the great sensitivity with which radioactive tracers are determined radioimmunoassays have been and continue to be widely used Use for the detection and determination of haptens and antigens.

Radioimmunoassays are carried out in such a way that the one of interest Mixes compound with a radioactive analog.

These two connections then compete for the bond to a limited amount of antibodies. The radioactivity bound to antibodies is made by the "free" radioactivity is separated and either the "bound" or the "free" Determined radioactivity. In a variant, a solid phase antibody is used, so that the unreacted "free" radioactivity is simply washed out can.

There are two closely related excess vine assay systems, the immunoradiometric assay (IRMA) and the 2-site IRMA. With these systems becomes a labeled antibody in excess used to the to convert an unknown substance into a directly detectable product. The non-implemented one labeled antibody is discarded. The 2-site IRMA is different from that IRMA in that the unknown substance during a previous reaction is made insoluble.

A wide variety of errors occur in immunoassays. Fluctuations the temperature and the reaction times can be critical. Most Systems are highly sensitive to errors in the separation of bound and "Free" marked connections (classification errors) and the errors caused by the Workstones such as the manufacture, measurement, and transfer of reagents (Pipetting errors) are always significant. Complicated assay variants in which sequential and / or non-equilibrated reactions are used particularly prone to failure.

The concentration of the compound of interest in an unknown Sample is determined by comparing the result obtained with the unknown sample with the results obtained when analyzing multiple solutions, in which the concentration of the compound is known (a standard solution).

It is therefore extremely important that all assays be carried out the same way be carried out with the same reagents.

One is constantly looking for a simple, precise procedure for performing immunoassays, whereby any deviations from the exact Work such as fluctuations in terms of response times, Differences in terms of the amounts transferred and the like., Are kept to a minimum and are common to both the standards and the unknown samples.

In addition, such a system should have a minimum number of operations include, in particular those which include an extraction, a dilution and a Include dispensing (dosage).

If necessary or desired, the system should be sufficient work fast and it should be able to show the results in terms of that accurately record the original sample. It should preferably be based on the determination of different materials such as small haptens and large proteins it should be feasible using both the RIA and IRMA procedures be the equipibrated (balanced) and consecutive Take non-equilibrated responses into account and apply in any given experiment effectively measure both large and small values. The procedures should also be economical and expedient. There is currently no automated measuring device that meets all of these criteria.

Various automated systems for measuring the most diverse Samples are described in U.S. Patents 3,469,438, 3,684,448, and 3,723,066. There are also a number of commercially available radioimmunoassay apparatuses. In addition belong the Centria of the company Union Carbide Corporation, the LKB-Produkter of the LKB, Sweden, and the Darius of Digico Ltd., England. Interesting in this context Articles are those of Miles, "La Ricerce Clin. Lab.", 5, 59 (1975), who himself concerned with immunoradiometric detection methods, Cottrell et al in "Clin.

Chem. 11, g32 829 (19W5), the computer-controlled automation of Radioimmunoassays refer to, Hersh and Yaverbaum in "Clinica Chimico Acta", 63, 69 (1975), and Jones and Worwood in J. Clin.

Path. ", 20, 540 (1975), who performed an immunorodiometric assay for Ferritin affects.

The present invention relates to a method and a Device for performing assays in which a representative of one of two families of a pair of specifically bindable (bindable) substances (SBS) in insoluble made form is used. Specifically bindable substances include the pair of ligand and receptor for the ligand. It is a receptor any organic compound or composition that can distinguish primarily due to a significant difference in the binding constant, between their conjugate ligand and other compounds, usually organic Compounds, with a similar structure.

In general, the ligands are haptens and antigens, and the receptors are generally antibodies (antiligands), although other proteins and polysaccharides are also used as receptors can. When using antibodies with conjugate haptens or antigens, these compounds are hereinafter referred to as immunologically bindable (bindable) Substances (IBS) called.

In the device according to the invention, a plurality of fixed Syringes used, the syringe plungers are connected to each other that they execute a synchronous movement. One electronically controlled Actuator is programmed so that the piston moves after a previously the established procedure. They are mechanical or electronic devices provided to any leeway (any phase shift) in the movement of the pistons balance when reversing their direction of movement. One arranged overhead Reservoir is preferably in fluid distribution and receiving relationship with the Syringes.

A complementary array of small, conical shaped tubes with open ends serves as syringe tips. The tips are delivered and stored and stored in a compact stack of interlocking tip support racks. The tips are made by pushing the tip ends down into the tips in the Carrying frame, in which the tips are carried by a special insert block, placed on the syringes. When performing the assay, the tips activated by making one of the representatives of an SBS pair insoluble in the tips power.

The tips can then be washed free of soluble material and stored or used directly. The solutions to be examined are held by a multiple sample holder introduced into the tips. The tips are dipped into the samples and the Samples are drawn into the tips by pulling back the plungers (plungers). The reagents can be drawn up into the tips or they can be done simultaneously introduced as part of the sample solution or introduced from the syringes. the Solutions in the tips can be ejected into a waste trough or into the syringes are pulled up, with all pistons moving synchronously, introducing solutions and at the same time solutions are expelled from the tips. To the versatile Usability in relation to reagent storage, new or used, can be increased even further an overhead reservoir may be provided, which through openings with the syringe barrels communicates. The reservoir can deliver washing solutions as needed can be inserted into the cylinders, or the reservoir can contain spent sample or absorb reagent solutions using mild suction.

After each operation or after using the various reagents the inside of the tips have come into contact and after all washing solutions are used Removal of any unbound material has been peeled off the tip ends and wiped the outer wall of the tips. Using radioactive tracers as a reagent and by using standards in the one to be measured at the same time Sample container can be used for direct determination of the unknown substance in the assay sample the remaining radioactivity in the tips can be related to the standard sample tips. The present invention therefore relates to a method for simultaneous implementation of several assays in which all stages of work are synchronized with the unknown solutions and the standard solutions can be carried out so that any introduced Errors for the standard solutions and the unknown solutions are constant.

Although the present invention is primarily to carry out of simultaneous multiple determinations, the method according to the invention can can also be used for individual determinations. By using a single Syringe and A single tip can make an assay (detection) quick be carried out by introducing and aspirating reagents and washing solutions by the tip, which allows the concentration of the samples, the handling of the The reagent is facilitated and suitable for the preparation of the radioactive sample for counting.

The tip is extremely versatile as it is extremely useful for sampling allows small volumes in very small containers, while at the same time a Surface supplies to carry out the various reactions necessary for the assay (Proof) are required. The inner surface of the tip can be used by of bumps can be modified to increase the surface area. In the top Spacer arms can be provided which then serve to insolubilize the analyte (the substance to be analyzed) or the reagent can be used.

Preferred embodiments of the invention relate to a method and a device for the simultaneous execution of several operations. The method and the device relate in particular to heterogeneous (two-phase) immunoassays, in particular the radioimmunoassay (RIA) and the immunoradiometric assays (IRMA and 2-site IR). These assays are primarily concerned with the determination of antibodies and immunologically active compounds (antigens and haptens). The device has a plurality of syringes which are mounted so that they are a simultaneous and allow identical movement of the syringe plungers. Also are facilities for fastening open tubes that are used as solid phase reagents, at the ends of the syringes for sucking in and ejecting unknowns Samples, reagents and wash solutions provided. In an upper section of the device a reservoir is provided for holding solutions or for distribution from solutions to syringes.

Single assays (individual detections) can be performed using a single syringe, the process steps being carried out in the manner that one transfers solutions through a hollow tube.

The invention is described below with reference to the accompanying Drawings explained in more detail. 1 shows a perspective view of FIG Syringe battery mounted in the motorized control mechanism; Figure 2 is a front elevational view, partly in section, of the syringe battery comprising is in contact with a sample plate; Fig. 3 is a partial elevational view in Cross section of a hollow piston; Figure 4 is a side elevational view of a syringe battery during storage; Fig. 5 is a front elevational view of the Figs Racks mounted tips; Figure 6 is a fragmentary perspective View of the assembled tips shown in Figure 5; and Fig. 7 is a front elevational view of tips mounted in a frame and carried by an insert block will.

To illustrate the present invention, the assay (Detection) first described in general terms, using immunoassays as examples and then it will be in relation to three different types of immunoassays, in which reagents containing a radioactive tracer are used explained. This is the radioimmunoassay (RIA), the immunoradiometric Assay (IRMA) and the 2-site immunoradiometric assay (2-site IRMA). After that, will then described the device according to the invention including the additional devices. The immunoassay is then applied to the specific device, with the Application of the device to the various immunoassays is demonstrated, then followed by specific examples in which the device for Determination of specific compounds of interest or

Compositions is used.

I. Assay Method The subject of the present invention is the implementation of assays with an SBS pair containing one ligand and one specific for the ligand Receptor includes. Pipes or

Open ended tubes used to facilitate introduction and removal of reagents, the spooling of the tubes or tubes to wash out of unwanted or redundant material and a light, reproducible one and allow simultaneous handling and precise final determination. In the widest Meaning the tubes or tubes are used in assays in which a representative a specifically bindable substance pair (SBS), e.g. haptens, antigens, antibodies, Enzymes, enzyme substrates, serum proteins and the like, inside the tube or

Tube is made insoluble.

A reagent is used which reacts to the insolubilized representative of the SBS pair is bound and is therefore itself a specifically capable of binding Represents substance that has been modified in such a way that it is detectable.

In Assoys, different atoms are used to carry out a measurement or compounds that provide the desired signal, such as radinactive Atoms such as iodine, tritium, carbon, and the like; stable free radicals; Enzymes; Enzyme cofactors; Fluorine-releasing substrates; Bacteriophages and fluorescent substances. The reagent, which is used for the assay determination is subsequently used as a detectant (detection substance) designated.

The distribution of the detectant between the solid phase and the liquid Phase is determined by the amount of the compound to be determined, the analyte that is present in contained in the sample.

By creating competition between the detectant and the analyte towards the insoluble representative of the SBS pair or by introduction of the analyte and subsequent Introduction of the Detectant into the Open-ended tubes will affect the amount of analyte present Amount of the detectant that has been made insoluble. Depending on the type of assay, either the amount of the detectant in the tube or in the liquid phase is determined or both can be measured. The tubes or tips, the one Critical roles in the present invention can play from the most diverse Materials exist. The tubes do not need to be made of a single material consist, the important aspect of the tube is the inner surface or the filling in the tube. The inner surface or a filling or both can be the surface result on which the representative of the SBS pair is made insoluble.

The tips can be made from a wide variety of materials, e.g.

made of polyethylene, polypropylene, polyvinyl compounds such as polyvinyl chloride, Polyacrylonitrile, polyacrylate, polymethacrylate and mixed polymers thereof, polystyrene, Nylon, polyterephthalate, cellulose and the like., Exist. That means it can be in the Naturally occurring polymers, preferably naturally occurring modified polymers, and synthetic addition and condensation polymers can be used. It can inorganic substances such as glass can also be used.

While usually only the inner surface of the tip is the active one Surface for insolubilizing the representative of the SBS pair the tip can be conical or partially blocked to small particles or Pearls of the above materials, regardless of or in Working with the inner surface of the tip as an active surface for insolubilization can serve. Alternatively, an uneven surface, e.g. one with bumps or protrusions, as an inner surface, e.g. with ribs, elevations or the like, be provided to increase the surface. The insolubilization can affect the can be achieved in various ways. Depending on the type of special surface as well depending on the type of representative of the SBS pair that is to be made insoluble the absorption or the adsorption can be satisfactory. In this way of working a solution containing the representative of the SBS pair is sucked into the tip, so that the representative can bind to the surface. After a sufficient Duration the solution is expelled and the inner surface of the tip becomes washed with appropriate media, usually buffered media. In suitable In situations the analyte from the unknown sample may be the insoluble made representatives of the SBS pair act from the sample on the inner surface the tip is absorbed. A labeled receptor can be used to detect the absorbed Analytes are used.

Another method is to get the representative of the SBS pair binds covalently to the surface. There can be spacer arms covalently to the surface are bound using various methods known per se, wherein a function remains that is either active or can be activated, so that she responds with the representative of the SBS couple. For example, nitriles can be modified to form Imidoesters available with Amino groups on an antibody or antigen react. In relation to the bond of proteins on surfaces using activated carboxylic acids, carbodiimides, Imidoesters, active alkyl halides and the like with the formation of amido, amidine or amino bridges, there is an extensive literature.

Alternatively, an immunological spacer can be used. A representative of the IBS pair is bound to the surface, who then has the complementary representatives of the IBS pair binds to the surface. Apart from the representatives of an IBS pair for binding belong to further materials themselves proteins that bind to plasma, tissue gates, enzymes, cofactors, substrates, inhibitors and the like, virtually any material that has specificity for the analyte.

The small conical tips that are commonly used have a lower opening with a diameter (I.D.) within the range of about 0.25 to about 2 mm, preferably within the range of 0.4 to 1 mm. The height of the tip is not critical and it can depend on the desired one Volume of the tip can be varied. Usually the height of the tip is about 1 to about 8 cm, preferably 2 to 5 cm. The top opening of the on the syringe The assembled tip is sized to fit snugly into the syringe barrel fits, and generally has a diameter (I.D.) within the range from about 2.5 to about 6 mm.

The thickness of the wall can vary, and it is for the present invention not critical. Appropriately, the thickness can vary along the wall, whereby they decreases with decreasing inner diameter of the cone. A range of thicknesses that the gives desired dimensional stability, is about 0.1 to about 1.5 mm, vorzugswpise at 0.2 to 1 mm, with the thickness ratio from the upper section (tip) to the lower Section (bottom) of the top can vary from 1: 1 to 10: 1. The volume of the tip is suitably within the range from about 50 to about 700 Rl, preferably within the range of 75 to 500 pl.

While generally the assays use combinations comprised of iBS pairs, there are special situations in which the interested Analyte has a specificity for a different substance than the other representative of the IBS couple has cases in which it is expedient to use the related bondable Substance to use. For example, if an enzyme substrate is to be determined, the enzyme and substrate can be used. When you specifically determine NAD If you want, you can make a tip in which the NAD is bound to the surface is.

When combining the sample with excess enzyme in the NAD as Substrate is used, and then introducing the combination into the Only the enzyme that was bound to the NAD in the sample will tip, not to the Lace surface bound. By ejecting the solution from the tip, one can remove the Determine the amount of enzyme that was not made insoluble.

The tips can be used immediately prior to performing the assay activated or they can be pretreated and stored. When storing the tips usually mild temperatures, generally from about -40 to +250 C, applied and the tips remain in a controlled environment to avoid deactivation of the insolubilized material. The inner surface can expediently the tips may be coated with different liquids to make a moist polar Maintain environment. Examples of suitable materials are polyols, such as Glycerin.

Depending on the type of assay, the unknown sample can be mixed with other reagents be combined or not. In most cases the sample will be more suitable Buffered in a way to improve binding, using common buffers and produces a pH that is within or near physiological range is in this area. Where the assay requires that the analyte to the detectant binds to the number of available binding centers that are insoluble can be made to affect the assay sample is usually below Conditions under which such binding can occur with the Detectant Reagent combined. If the detection is made insoluble to such an extent, until the analyte has been rendered insoluble, the analyte normally becomes inserted into the tip first, then the detectant reagent is introduced. To any introduction of a solution into the top is usually the top of any Washed free materials that are non-specifically bound to the surface.

Different incubation stages can be used in which the tips stored at ambient or controlled conditions usually at a temperature within the range of 15 to 400C and with a humidity that prevents drying. Usually be the tips after the introduction of the analyte and when the detectant is introduced separately is incubated after the introduction of the detectant. The incubation times can vary from 0.5 to 12 hours, depending on the particular compositions used, the concentrations to be detected and the like.

The method according to the invention is particularly advantageous in use on dilute analyte solutions. By appropriate selection of the assay method, you can large excesses of the insolubilized representative of the SBS pair were used will. The analyte solution can then be sucked through the tip, whereby the Analyte is made insoluble by the excess insolubilized agent, which is bound to the surface. A controlled rate of suction of the solution through the tip and into the syringe can be proportionate lead to rapid quantitative removal of the analyte.

In order to explain the method further, the following are general detailed three different assays using rcdioactive iracers. The first assay discussed is RIA. At the RIA, going out that the antigen is the analyte, a competition between one radioactive labeled antigen (detectant) and the antigen analyte generated. A limited one lot Antibody to the antigen is bound to the inner surface of the tip. the unknown antigen solution is then introduced into the tip, either at the same time with the detector or before the introduction of the detector. The advantage of the introduction of the two solutions in succession instead of in the form of a single solution is the one that one can tip off any interfering materials in the unknown sample can wash freely before the leg of the detector. In addition, if the ligand and the receptor are in a rapid state of equilibrium, the detectant and the insolubilized representative of the SBS pair are preincubated, before they are combined with the analyte. This makes it possible to use the reagents in a convenient and reproducible form and those attributable to the serum To further reduce fluctuations.

The next assay procedure to be discussed is the IRMA.

In the IRMA, the unknown analyte is combined with a reagent, contains or consists of soluble, purified radioactive antibodies (detectant). The tip has an insolubilized analyte or a homologous analyte on.

The combined analyte and antibody solutions are in the top sucked in, in which all free centers of the antibodies to the insolubilized Representatives of the IBS couple are bound. There is a change to the IRMA system in using antiglobulin (anti (antiligand)) that is radioactively labeled (Detectant). In this embodiment, the antibodies are not to the analyte marked. After the analyte antibodies have been made insoluble in the tip, can then the radiolabelled antiglobulin inserted into the tip so that the labeled antiglobulin is bound to the analyte antibodies, which have been made insoluble.

The third method discussed here is 2-site IRMA. In this procedure, antibodies become insoluble for the analyte in the tip made. The analyte is then introduced into the tip and attached to the antibody bound. The analyte must be polyepitopic, i.e. it must have at least two binding centers have which the simultaneous binding of at least two antibody molecules allow. After the analyte is bound to the insolubilized antibody, unreacted antibodies and other solutes are washed out the radioactively labeled antibody (detectant) for the analyte is in the tip introduced, in which he attached to any existing insolubilized analytes is bound. Alternatively, as described for FIRMA, you can use labeled antiglobulin Use (detectant) instead of the labeled anti (analyte) so that the labeled Antiglobulin can be used regardless of the particular analyte.

In all assays (detection methods) the solution is used after the introduction of the labeled detectant into the tip and after insolubilizing it the tip ejected or withdrawn, the inside of the tip with a suitable Solution, usually a buffered solution, is washed to any Remove non-specifically bound material, and the ends of the tips will be wiped and extinguished. This will get any residual liquid in the end the tip and any material on the outside of the tip removed.

II. Device The device can be divided into three basic parts are: (1) the syringe battery, (2) the electronically controlled motorized one Drive and (3) the auxiliary equipment. The syringe battery is next first explained.

Syringe battery The syringe battery has a large number of evenly Syringes arranged in rows and columns in a housing, the syringe barrel or syringe barrels are attached between two platforms, the ends the syringe barrel extend beyond the lower platform. The upper Platform may have a hollow portion that serves as a reservoir, wherein each of the syringes has one or more openings that are in fluid communication or is in fluid communication with the reservoir. Movably attached to the frame and arranged below the lower platform is a plate through which the the tip-receiving ends of the syringes extend which are pushed down can be used to remove the tips from the syringe tip receiving ends, and which can then be withdrawn so that new tips can be placed on the syringe tip female ends can be attached. A connection device is provided, which connects the plurality of pistons together, the pistons being complementary are arranged to the syringe assembly and into the syringe barrel extend. The plungers are long enough to be the length of the syringe barrel traverse and remove all material from the cylinders eject can. The pistons can be solid or hollow and a spring valve, which the Introducing solutions into the syringes through the plunger allowed to have.

Electronically controlled motorized drive The syringe battery is mounted in an electronically controlled motorized drive that is shaped like this is that it houses the syringe battery and an upper platform for the housing the syringe plunger plate. The platform is mounted for vertical Movement with simultaneous movement of the syringe plunger plate. It is a lower one Carrier element provided, which holds the sample cells or reagent cells, which can be raised to the top and then lowered again. Than less A suitable alternative is the syringe battery up to the sample platform lower. For rigidly holding the lower plate in the desired position during locking and support devices may be provided for the various operations be. Drive devices can be used to raise and lower the upper and carrier elements be provided which can be electronically controlled and programmed so that the assay can be performed essentially automatically. During the assay it may be necessary to replace the sample cells.

Additional equipment A top rack is provided for which it is a plate with a multitude of holes that are complementary to the distribution of the syringe assembly. It is useful in addition to the holes for the tips a variety of smaller holes provided, which have a similar distribution as the larger holes between the larger holes to have.

In this way, the syringes with the smaller ends of the tips, that are contained in the smaller holes are stacked so that one staggered (staggered) configuration is obtained, creating a large number of top plates one on top of the other can be arranged. This way, a large number of tips can be placed on one relatively small area can be stored.

A second piece of the additional equipment is subsequently called an insert block designated. The block has a multitude of complementary shaped chambers for the inclusion and retention of the smaller ends of the tips, which is a mechanical one Form supports for the tips. The insert block is used when the tips can be mounted on the tip-receiving ends of the syringes by turning the Syringes into the tips carried by the block, a third piece of the Additional equipment is referred to as a storage carrier box, which is a moisture-controlled Environment supplies which, if desired, can also be kept at the same temperature can.

For a better understanding of the device according to the invention this explained in more detail below with reference to the accompanying drawings.

A multiple assay device 10 has a motorized housing 12th and a syringe battery 14 as shown in FIG.

The motor-driven housing 12 has a support frame 16, below to which a motor 20 is attached, which is electronically controlled by a programming device 22 is controlled. On the platform 12 are four threaded screws or bolts 24 are rotatably fastened, which are seated in gears 26.

The central gear 30 is driven by the motor 20, and it drives the gears 26 at the same time.

An upper platform 32 is by means of a screw thread on the threaded screws or bolts 24 attached so that they have a vertical Movement while turning the screws or

Bolt can perform. The upper platform 32 is symmetrical arranged L-shaped arms 34 and 36 for containment and secure retention of the syringe plunger plate 40. On the platform 12 are opposite one another Sides of the syringe battery 14 side walls 42 attached. Extend from the walls shoulders 44 inside, which carry the syringe battery and the syringes Lock the battery in the desired position. From shoulders 44 outwards extend guides 46 on which a lower platform is movably mounted is. The lower platform is mounted on a cam (not shown) that a cam arm 52 for raising and lowering the lower platform. feathers 54 are mounted on guides 46 to retract the lower platform.

On each of the guides stops 56 are provided to the lower To hold the platform above the middle gear 30.

There are safety switches 60 and 62 provided to the degree of to control vertical movement of the upper platform 32.

In the support bracket 66, a stand 64 is attached to the safety switch 60 to control the degree to which the upper platform 32 will be lowered can take up. In the support bracket 66, a stand 70 is attached, the one Arm 72 has the safety switch 62 for controlling the degree of upward movement the upper platform 32 receives.

The two normally closed safety switches 60 and 62 are in communication with a control mechanism 22 during operation so turn off the motor 20 when the switches are turned on.

The syringe battery 14 will now be described with reference to FIG Fi. 2 and 4 explained in more detail. The syringe battery 14 has a rectangular housing 70 with an upper guide plot 72 which is fastened to the housing cover 74 is. The guide plate has a plurality of channels for guiding the pistons 76 on.

The cover 74 can have similar channels or it can be hollow and serve as a reservoir for a solution, in particular a washing solution. This is explained in more detail below.

The support rods 80 support the cover 74 and are on a base plate 82 attached, which in conjunction with the cover 72 serves to hold the syringes 84 to hold in the desired position.

The bottom plate 82 is made rigid by side walls 42 and shoulders 44 held in the desired position. The tip receiving ends 86 of Syringes 84 extend through and beyond the bottom plate 82 in a sufficient Length so that the tips 90 are at the syringe ends 86 can be attached. The syringe plunger plate 40 has a hollow center 92 for receiving the piston heads 94 and for holding the piston heads in one fixed position.

The ejector plate 96 is attached to rods 100 at its four corners. At each end of the syringe battery 14 are rods 102. Springs 104 are open the rods 100 are fixed between the rod 102 and the bottom plate 82 so that they withstand the upward pressure against the rod 102. The cam 106 is rotatably mounted on the pin 110 in the yoke 112 and is by means of the cam arm 114 moves. By rotating the cam 106, the ejection plate 96 is moved downwards, as shown in phantom at 116 so as to separate tip 90 from the tip receiving Remove ends 86 of the syringes. The rods 100 extend upward through the cover 74 so as to remain aligned and not bound during movement to become. The tips 90 are in a rack 116 for storage and during for use in the assay device 10. The frame 116 allows the simultaneous fastening of all tips on the ends receiving the tips 86 and the simultaneous repulsion while they are held in the arrangement, in which they originally existed. A sample or reagent cuvette 120 is provided, the complementary beaker for the delivery of reagents to the tips or the intake of reagents aspirated from the tips.

It is convenient to use the manual operations during the assay or keep an operator's attention to a minimum.

Therefore, it is desired that the solutions, reagents and the like are automatically can be inserted into the tips 90.

In FIG. 3, the cover 74 is hollow with a reservoir 118 shown. The reservoir 118 stands through the opening 122 with the syringe 104 in fluid connection (fluid connection).

When the piston 76 is moved across the openings, liquid occurs from the reservoir into the syringe 104, with a positive pressure on the reservoir exercising as necessary to force the liquid into the syringe and this can then be expelled from the syringe 76 into the tips 90 by the piston is moved downwards. Alternatively, a hollow piston with a central one Stand 124 is used, which is mounted on a spring and connected to the foot 126 An O-ring 130 is placed between the foot 126 and the protrusion 132, which is rigidly attached is attached to the piston 76, compressed. The foot 126 is shaped so that between a gap remains between the foot 126 and the inner wall of the syringe 76. The O-ring 130 creates a seal with the inner wall of the syringe 84 by exerting a Pressure on the central post 124 lowers the foot 126, so that a fluid connection or fluid communication between the hollow center of the plunger 76 and the syringe 84 is created. The liquid (the fluid) can therefore be distributed by means of the piston or withdrawn from the syringe by suction. These types of pistons are commercially available and are called 'Eppendorfer flasks;' designated.

Additional devices The frame 116 has already been explained as a device for the Arrangement of the tips 90 at a distance from one another accordingly the syringes 84. The racks are also used for the storage and transport of the Sharpen. In addition, as shown in FIGS. 5 and 6, the frame can have a plurality of small holes 134 which are symmetrical in the same way as the larger, the tips retaining holes 136 are arranged. By using These two types of openings allow the tip to be stacked one on top of the other make sure that the small holes line up with the tips in the rack above are and wear them.

A tip injection block 140, as shown in FIG. 7, is also desired. When attaching the tips 90 to the tip receiving ends 86 of the syringes 84 the tips are arranged in conical bores 142, which have a mechanical Represent carriers for the tips. Doing this can create considerable pressure applied to the tips to ensure that the tips are firmly attached to the Syringe ends are attached. The tips 90 are small, their wall thickness is sufficient however, it is sufficient to impart considerable mechanical strength to the tips.

Explanation of the implementation of an immunoassay For further explanation of the subject matter of the present invention is subsequently a multiple assay which was performed in the multiple assay device. For explanatory purposes a 2-site IRMA assay is used.

A plurality of tips 90 are arranged in a tip rack 116. the Holes in the racks are complementary to the arrangement of the syringes 104 in the syringe battery 14. Not all of the holes need to be used, as the method and the Device does not require all syringes to be in the syringe battery be used.

Usually standard solutions of the analyte and the samples are used prepared, where any dilutions as required can be carried out. the Various standard solutions and assay samples are placed in the beakers of an assay container 120 introduced.

The syringe battery can expediently be used to prepare the various samples 14 can be used to dilute the samples, add the reagents and the like. By using the syringe battery 14, all operations are identical to the Operations that are used with the standard solutions.

The tips 90 attached to the frame 116 will now be inserted into the insert block 140 inserted, the syringe ends 86 inserted into the tips and the syringe battery 14 is pushed down to cause the syringe ends to be firmly attached to the tips sit.

The syringe battery 14 is then inserted into the motorized housing 12 introduced by sliding the syringe plunger plate 40 between the L-shaped Arms 34 and 36 of the upper platform 32.

A sample cuvette with the appropriate antibodies is placed on the lower Platform 50 placed, with the upper platform 32 in their lowered Position is so that the pistons follow a considerable distance extend down into the syringes.

The syringes 84 can expediently be partially filled with washing solution so that after the antibody has been drawn up into the tips 90, the tips directly can be washed after expelling the antibody solution by propulsion the plunger down and by expelling the vial solution through the tips.

After a sufficient period of time, namely after about 1 to about 16 hours, the contents of the tips are placed in a waste container (a waste cuvette) ejected and the tips are washed twice, either with a washing solution in the syringes 84 from the upper reservoir 118 or with a wash solution in one Sample container (sample cell). After completing each work step, the tips wiped and extinguished. The tips 90 are then by means of the ejection plate 96 ejected and they can be frozen or in a simple storage container 144, as shown in FIG. 4, can be stored under controlled humidity.

The carrier control container 144 is rectangular Box 146 without a cover that carries the syringe battery 14, with the syringe battery used to enclose the box. A damp sponge 150 is included in the box, which creates the desired moisture so that the tips do not dry out.

Assuming that the tips 90 are retained on the syringe ends 86 the samples and the standard solutions containing The cuvette (the container) is now placed on the lower platform 50 and raised, like this that the tips substantially reach the bottom of the cups 152 (see. Fig. 4).

The approximate amount of the solution is then drawn up into the tips and the unit is either left in the motorized housing 12 or in the Bearing carrier boxl44 transferred. The incubation of the samples is under controlled Temperature and humidity conditions.

The tips are then washed as indicated above, one The reagent cuvette containing radioactive antibodies is placed on the lower platform 50 and raised so that the tips are essentially over the entire Distance of the cups 152 extend. The reagent is released by lifting the top platform 40 drawn into the tips and after the antibody has been given enough time for the Has given binding, the labeled antibody solution is expelled into a waste cuvette and the tips are washed, wiped and blotted dry0 In the syringes A variety of reagent solutions can be contained and these can be gas bubbled be separated from each other.

In this way you can go through a number of different solutions the tip is carried out by sucking reagents into the syringes several times the sample cuvettes, whereby the aspiration is carried out alternately in the air. The amount of clearance air should be sufficient to expel the solution from the tip.

These used up solutions can be discarded by getting them withdraws into the reservoir.

The tips are then ejected again into the insert block, wherein however, they remain in contact with the tip rack 116.

The tips are then inserted one after the other into counting tubes for counting the radioactivity adhering to the inner wall of the tips. The amount of radioactivity at the tips is a direct function of the amount of in the standard solutions or analytes present in the unknown solutions. By making a standard curve you can compare the results with the unknown samples with the standard curve, to calculate the amount of analyte in the unknown samples. In a similar way other immunoassays can be performed according to the applicable procedures, such as RIA, IRMA, enzyme immunoassays and the like.

The device according to the invention has been described above with reference to the Use in one immunoassay is explained in more detail, but it can also be used in all others In cases where it is desired to have multiple simultaneous transmissions of liquids, so that the device according to the invention for the automatic pipetting, for automatic sampling, for automatic Suction or the like. Can be used.

The invention is more preferred in the following examples by hand Embodiments explained in more detail, but without being limited thereto. All in it specified temperatures are given in ° C and all percentages relate to unless otherwise stated, by weight.

Example A Use of an assay device to perform the 2-site immunoradiometric assays (2-site IRMA) for the determination of human plasma growth hormone (HGH) Procedure 1 1. Preparation of the standard solutions and unknown samples Es Standard solutions of HGH were prepared with hormone-free human blood plasma that had been diluted with a diluent in the ratio of 1: 5 (Diluent = 0.05M sodium barbitan, 0.1M NaCl, 0.1% bovine serum albumin (BSA) and 0.01% sodium azide with a pH of 8.0; this diluent is hereinafter referred to as BSA buffer * Four replicates of each of the ten standard solutions (o to 10 ng / ml) were made stored in a plastic container with 96 positions (Cooke Microtiter Ltd.). The container also contained 6 replicas of each of the 3 "quality control" human plasmas, which had also been diluted in a ratio of 1: 5. These human plasmas (Q1, Q2 and Q3) were selected to have low, medium and high levels of circulating HGE and they were frozen in the form of many aliquots.

Human plasma samples with an unknown HGH content were tested with a Diluent diluted in a ratio of 1: 5 and analyzed in duplicate. the samples analyzed at a dilution of more than 1: 5 were compared with the in the ratio 1: 5 diluted hormone-free human plasma further diluted. After each 44 "unknown" Tips were used a duplicate "zero dose" standard and duplicate Q2 samples These assay samples were made and diluted in the containers using the same test device as in a sample diluent. the Samples were stored in the frozen state in the containers after using a Cookes M microtiter distributor and tape.

2. Making the Tips Each tip rack was made using of the insert block placed on a unit. The top support frame stayed with connected to the tips. The tips were drawn up by drawing up 120 / ul Neerschweinchen-Anti (HGE) serum, diluted with 0.2M sodium bicarbonate of pH 9.2 in a ratio of 1: 4000 "covered" with anti (HGH). This antiserum was raised from one under the tips Cell (bowl) drawn up. After 1 to 16 hours, the contents of the tips became ejected into the waste cuvette and the tips were twice with the liquid washed from the wash reservoir. In each case 120kl1 washing solution was put into the waste cuvette given. The washing solution contained a 0.05M phosphate buffer of pH 8.0, 0.1% BSA, 0.45 ffi NaCl and 0.01% NaN3. At the end of each pass, the tips were wiped off and extinguished.

3. Storage of the activated tips Alternatively, the 120 "coated" antiserum or tips containing washing solutions were frozen and the tips became complete ejected from the tip support frame (using the tip ejection mechanism), and in the form of a stack of lace support racks in a plastic bag in FIG stored frozen. These tips were thawed and put back on the unit put on if this was necessary to perform the assay.

4. Assay method (detection method) (a) Incubation of an unknown Sample and a Standard Sample (Reaction 1) The assay began as the standards and the unknown samples into the "coated" and washed tips simultaneously were raised. 120 pl solution were under a programmable electronic Control raised using a minimal top airspace. The unit was then left in the motorized section or taken out and into the Bearing carrier box provided. In each case, the incubation was at a controlled rate Temperature and humidity performed, (b) First wash The tips were then washed, wiped, and bled in the same manner as above.

(c) Second reaction 120F1 were purified radioactive anti (HGH) then drawn up from a reagent cuvette. This reagent was in the BSA buffer diluted so that 120 »11 = 20,000 to 40,000 cpm.

The unit (plant) was incubated again as indicated above.

(d) Final wash. After 24 to 48 hours, the labeled antibody became ejected into the waste cuvette and the tips washed again, wiped off or extinguished to dryness.

5. Determination of the radioactivity at the tips The tips were ejected into the insert block using the ejection mechanism (where they were still attached to the tip support plate). Then the tips became one by one Introduced into counting tubes to count the number of sticks on the inner wall of the tips Radioactivity using an automatic gamma counter.

6. Data Analysis The amount of radioactivity at the tips is one direct function of the amount of IifGH contained in the standard or unknown solutions, From the results of the assay of the HGH standard solutions, a "standard curve" and the concentration of HGH in the unknown plasma was calculated by comparing the tube radioactivity of the unknown solutions with the tube radioactivity of the standard solutions.

Method 2 Use of the assay device to perform the 2-Site IRMA to Determine Plasma HGH by Washing Syringes This procedure was identical to procedure 1, but this time the syringes with a washing solution were filled. The first and second responses were with the unknown or labeled antibody, which was separated from the wall of the skin by an air bubble, carried out.

All solutions contained a small amount of propylene glycol for reduction the surface tension. At the end of each reaction, the solid phase was purged the solution in the tips washed into the waste cuvette, whereupon 200 µl of wash solution followed, ejected from the syringes and through the tips became. By means of the syringe piston, a chamber around the syringe barrel around, or by drawing new buffer into the syringes from below to be introduced.

Method 3 Use of the Assay Device to Perform the 2-site IRMA for determining plasma HGH with all of the solution in the syringe This procedure was identical to procedure 1, but this time however, upon completion of each reaction, the solution in the tips through the tips has been drawn into the syringes. There was a bubble gap during the reaction 2 required to remove the waste solutions (in the syringes) from the labeled antibody (in the tips) to separate. All solutions contained a small amount of propylene glycol to reduce surface tension. The waste solution in the syringes could through the plungers or through a chamber around the syringe barrel be removed with a mild suction effect.

Method 4 Use of the assay device to perform the 2-site IRMA to determine HGH with repeated aliquots of the unknown Samples This assay procedure was identical to that described in Procedure 1, this time, however, assay reaction 1 taking repeated Aliquots of the unknown sample (e.g. from 120 81 at hourly intervals) was carried out. When a new aliquot was taken, the previous one Aliquot drawn into the syringe. Alternatively, the previously removed Aliquots are ejected into the waste cuvette. This procedure yields an assay with increased sensitivity.

Method 5 Use of the Assay Device to Perform the 2-site IRMA for the determination of HGH with slow continuous suction of the unknown sample This assay procedure was identical to that described in Procedure 1, this time assay reaction 1 by converting an unusually large amount the unknown sample was performed. The unknown solution became slow and continuously sucked through the tip into the syringe.

In this procedure, the unknown sample was extracted from a larger one Volume reduced for reaction with the labeled antibody (reaction 2).

Example B Using the Assg Apparatus to Perform the IRMA In the IRflA method, the unknown antigen was purified with soluble radioactive Antibodies implemented. The radioactive complex remained in solution while unused radioactive antibodies removed by a second reaction with a solid phase antigen became. The amount of in solution remaining radioactivity was a direct function of antigen concentration. The IRMA was also on use of labeled anti-IgG applied as an additional "universal" reagent, whereby the need for the production of radioactive antibodies for the unknown Antigen specific was avoided. The assay for determining HGH taking Application of the IRMA was carried out in a manner analogous to that described for RIA and 2-site IEMA Procedure carried out.

In the IINA procedure, excess antigen became on the solid phase (in the tips) fixed and the unknown antigen and purified labeled specific Incubation mixture containing antibody was aspirated into the tips.

Any labeled antibody that does not bind to soluble antigen was tied to the tips. The tips were washed and the marker tied hint: was inversely related to the amount of in the unknown solution present antigen.

Example C Use of the Asssy device; to carry out the BLA joined the RIA procedure and the analogous sonic binding assay procedures a competition between the unknown antigen and the labeled antigen derivative with respect to a limited amount of a specific binding reagent (usually of an antibody). Free labeled antigen was linked to an antibody (or another specific binding agent) bound antigen using the different procedures separately, but if the binding reagent in if a solid phase was present, then the unreacted labeled antigen could simply washed out. The amount bound to the binding reagent labeled antigen was an inverse function of the concentration of the unknown Antigen. The assay could be performed in the form of a single reaction or the labeled antigen could later turn into a "delayed" or "second" response be admitted. This operation is often called "delayed addition of the tracer" and it often leads to increased assay sensitivity. If the The solid phase was washed free of unreacted, unknown components of the solution before the "tracer" (labeled antigen) was added, then joined the labeled antigen no potentially impairing or interfering components the unknown solution directly in appearance.

In an alternative method, the labeled derivative (detectant) could are reacted with the solid phase antibody (the insoluble representative), before the reaction with the analyte was carried out. With hapten assays it was in this way possible to carry out reproducible pretreatments without the dose response was affected, and in this way could Assay interference can be prevented by the presence of serum.

Example D Use of Assay Device to Perform RIA for the determination of CortisoL in human plasma method 1 1. Manufacture of standard, unknown and quality control samples Nine quadruplicate standard solutions (with 0 to 5000 ng / ml) were processed into a solution containing 0.05M phosphate buffer pH 8.0 containing 0.1% bovine serum albumin and 0.01% sodium azide. This solution is referred to below as BSA buffer. Quadruple quality control plasma samples became (Q1, Q2 and Q3) used which show the low, medium and high levels of circulating PlasmacortisoL represented. The unknown plasma samples were duplicated examined. The 125J-CortisoL was diluted with BSA buffer.

The standard samples, the quality control samples, and the unknowns Plasma samples were diluted 1:21 with 95% ethanol. The one with it The resulting precipitate was centrifuged off and an aliquot of the supernatant Fluid was diluted with 125J-cortisol. The final sample dilution was 1: 399 and it contained 6000 cpm / 100Fl10. This solution was stored in sample plates and covered with a special film until the assay procedure is carried out.

2. Pre-treatment of the tips The tips were as indicated above activated, but this time using multiple layers of IgG. on an initial coating of normal rabbit serum diluted 1: 1000 with 0.2M NaHCO3 (pH = 9.2) a coating of guinea pig anti (rabbit IgG) serum, diluted with a BSA buffer in a ratio of 1: 1000.

After 4 hours this solution was expelled and the tips became washed twice with the BSA phosphate buffer. The tips could now be fed Bind (insolubilize) antiserum produced in rabbits. By doing The CortisoL assay was diluted 1: 25,000 with a BSA buffer Rabbit anti (Cortiso) serum sucked into the tips and pre-drawn for 16 hours incubated after washing in the BSA buffer. All incubations were at ambient temperature carried out.

The tips could be saved for later use, wherein they were frozen in racks while they were still antibody solution or BSA buffer contained. The tips that were stored before the finished specific Antibodies had been converted could be used for any assay using a rabbit antiserum.

3. Assay procedure The solutions to be tested (which also contain 125J-cortisol contained) were drawn into the tips from the sample cuvette. The incubation was kept at room temperature in the inotorized section or in the bearing carrier box carried out at controlled temperature and humidity. After 16 hours were the tips evacuated, washed and dry blotted before being attached to the solid phase bound radioactivity was counted (alternatively, the 125J-CortisoL not mixed with the unknown sample, but later in a second reaction, d. H. in the form of a "delayed addition of the tracer" or in the form of a pre-incubation of the tracer "can be added).

4. Dose Integration The amount of radioactivity bound to the tips was an inverse function of the amount in the standard solution or in the unknown Solutions existing OortisoLs.

Method 2 Use of the assay device to perform the RIA for the determination of CortisoL using the delayed addition of the tracer and using repeated aliquots of the unknown Sample This assay was performed as indicated above, but this time it was repeated aliquots of the unknown sample were taken and the unknown sample was not mixed with 5J-Cortisote. The 125J-CortisoL was in the form of a second (later) reaction raised. This resulted in an assay with increased sensitivity.

Method 3 Use of the Assay Device to Perform the RIA for the determination of CortisoL using the delayed addition of the tracer and slow continuous aspiration of the unknown sample. This assay was used carried out as stated above under method 2, but this time the unknown Sample was slowly and continuously drawn into the syringes through the tips (as described for the 2-site IRMA). This resulted in an assay with an increased Sensitivity.

When using method 1 for determining human growth hormone The following results were obtained in hormone-free human plasma: Table reproducibility the assay device 2-site IRMA for the determination of human growth hormone in hormone-free Human plasma diluted with a buffer in the ratio 1: 5 - CV of the sensitivity (Radioactivity) via the standard curve and the quality control plasma, and the minimum detectable dose (MDD), incubation volume = 120µl.

HGH concentration% CV = SD / mean x 100 mean CV of response (ng / ml) Assay number sensitive- (replicates sensitive- = 4) 1 2 3 4 5 6 speed 0.0 5.90 1.15 3.74 6.51 5.07 4.10 4.41 * 0.02 6.40 2.48 2.86 1.78 2.22 4.58 3.38 * 0.05 4.20 3.22 2.97 4.91 3.09 3.70 3.68 * 0.1 2.80 0.98 2.00 1.06 2.53 2.16 1.92 0.2 0.73 4.46 2 , 27 1.78 2.82 1.34 2.23 0.5 1.23 2.41 0.62 2.56 3.04 0.51 1.73 1.0 1.55 2.24 1.52 3. 69 1.44 1.87 2.02 2.0 1.18 2.75 1.24 1.40 0.93 1.51 1.50 5.0 3.63 1.02 1.37 1.42 3, 53 1.60 2.09 10.0 2.45 0.85 0.82 0.81 0.96 0.62 1.08 Table (continuation) HGH concentration% CV = SD / mean x 100 mean CV ng / ml) assay number of Response replicas assay number sensitive- = 4 1 2 3 4 5 6 quality control plasmas (Replicates = 6) mean 0.11 ng / ml 2.28 5.50 2.03 2.14 2.50 3.37 2.97 mean 0.36 ng / ml 0.77 1.73 3.59 1.32 2.14 2.35 1.98 Mean 1.70 ng / ml 1.23 2.24 1.63 2.45 1.29 1.70 1.75 Total mean CV of responsiveness (all points) 2.36 mean excluding the three lowest standard samples 1.93 MDD one Double determination mean (picograms / 120µl) MDD 1.43 0.83 1.06 0.95 0.40 0.40 0.84 * The responsivities were 800 to 2000 cpm, so that the counting error for 3.5 in this part of the standard curve was 2.2% (counting time = 1 minute).

When using methods 4 and 5 of the 2-site IRMA method described above greatly improved sensitivity was achieved. In the standard curve was the cpm spread of tube radioactivity between 10 and 100 pg / ml for the procedure 1,300 cpm. Method 4 had a spread of about 1200 cpm, while method 4 had a spread of about 1200 cpm Method 5 the dispersion was approximately 1250 cpm. In addition, the curves pointed at that Method 4 and in method 5 in comparison to the curves in method 1 much steeper drop in the 10 to 50 pg / ml range. Therefore, depending on the desired accuracy and sensitivity extremely low concentrations easily be detected.

The method according to the invention is extremely simple and sensitive Process for the determination of the most varied of immunologically bindable substrates or other compounds for which there are known receptors. The manual Operations are minimal, especially in areas where individual Errors can occur. Since all transmissions are simultaneously calibrated with carefully and equivalent spraying, all conditions affect the as a function of time, the standard samples do not fluctuate differently from that unknown sample. Therefore the standard determinations and the standard curve provide Results under conditions identical to those under which the unknown samples were determined.

The inventive method is in a tube with an open Carried out at the end so that solutions can be easily introduced and withdrawn, without movement or disturbance of the tube. Unlike one simple test tube does not occur with the open-ended tube mechanical Disturbance during the withdrawal of the solutions.

In addition, a column effect can be achieved in the open tube, so that the unknown solution becomes the new, non-unsettled, insoluble one SBS can be introduced into the tip by drawing up the solution without equilibration (Equilibration) between the insolubilized SBS and the analyte he follows. The reagents and / or washing solutions can be poured into the tips from the syringes can be introduced and later ejected into a washing level. The reagents and / or wash solutions can be transferred from the reservoir via the syringes into the syringes to be introduced. Alternatively, the reagents, wash solutions, and assay sample inserted into the tip from a cuvette and later drawn into the syringes will. The "used" samples, reagents and / or washing solutions can be passed through Suction through the reservoir can be removed from the syringes. If necessary successive solutions can be kept from one another by keeping air bubbles apart be separated.

The device according to the invention creates a convenient method with whose help pre-treats a large number of samples in the cuvette, the samples in the pretreated tips introduced and the multiple determinations at the same time and can be carried out under identical conditions. Using Carefully calibrated and same syringes and plungers can be extremely accurate and reproducible Results are achieved. Large or small volume samples can be used will result in effective insolubilization of the analyte the great capacity in the tips.

The tips are extremely versatile in terms of the role they play to play. By using conical tips with a small opening and a conical outer diameter at the opening allows the tips for sampling and used for aspirating extremely small volumes, making a more efficient Transport from the sample cell to the tip is possible. The tip can go into the Sample can be immersed without overflowing. The volume of the tip is sufficient to complete the various reactions required to perform the assay are to be carried out and you can then go straight to the determination of the detectant be used.

In the devices used so far, a single sampling device was used used. It could happen that material contained in a sample was in the sampling device be deposited, causing all subsequent samples to fail. at the device according to the invention prevents such an error from occurring.

In addition, the original arrangement of the samples and standard solutions, if used, maintained throughout the assay so that each sample is a specific one Position, thereby recording the results, especially on automatic Ways that is facilitated. Finally, in many cases, tips can be used with all reagents in the tip can be prepared beforehand so that measurements of the reagent volumes are carried out The techniques used in the laboratory are avoided, thereby reducing the introduction of human Errors are practically excluded.

The invention has been described above with reference to preferred Embodiments explained in more detail by way of example, but it is self-evident for a person skilled in the art that it is by no means restricted to this, but that it is in many respects altered and modified without departing from the scope of the present Invention is abandoned.

Claims (31)

Device and method for multiple analysis Patent claims f Method for performing assays for the determination of the amount of an analyte in a sample in which the analyte is a member of one of two families used, which consist of a pair of specifically bindable substances, and in the assay one insolubilized representative of this pair as well as uses a detective who is a representative of one of these families acts and that depending on the number of molecules present Detectant supplies a physically measurable signal, which means that that a solution of the analyte by means of a syringe into a tip with an open End draws, which is detachably attached to the syringe and which is insoluble contains made representative, with the proviso that if the Analyte can be bound to the tip, while the analyte is insoluble made representative acts, solutions of the detectant or other reagents and Wash solutions from an external source or from the syringe by means of the syringe into the tip, the solutions removed from the tip using the syringe and, by means of a physically detectable signal, the amount of that which has been insolubilized Detectants in the tip or the detectors removed from the tip as a measure of measures the amount of analyte in the sample. 2. The method according to claim 1, characterized in that it is the assay is an immunoassay and that the pair of are specific substances capable of binding are ligand and antiligand. 3. The method according to claim 1 or 2, characterized in that it the analyte is the ligand that it is the insolubilized one Representative is the anti-ligand that is present in insufficient quantity, to bind all of the ligand and all of the detectant, and that it is at the detectant is a radioactively labeled ligand. 4. The method according to claim 1 or 2, characterized in that it the analyte is a polyepitopic ligand that it is the insoluble representative is an anti-ligand and that it is in a Amount in excess versus the amount of ligand present is present and that the detectant is a radioactively labeled antiligand acts. 5. The method according to claim 1 or 2, characterized in that it the analyte is a polyepitopic ligand that it is the insolubilized representative to an antiligand of a first group of Connections and that it is the detector. a combination of an antiligand from a second group of compounds and the anti (antiligand) for the second group of compounds. 6. The method according to claims 1 to 5, characterized in that that the insolubilized representative is a ligand which is present in an amount in excess compared to the amount of the detectant present, that the detectant is an antiligand that is simultaneously with the analyte is introduced into the tip. 7. The method according to claims 1 to 5, characterized in that that the insolubilized representative is a ligand which is present in an amount in excess of the detectant present that it the analyte is a polyepitopic ligand and that it is in the case of the detectant, a combination of an antiligand from a first group of compounds and a radioactively labeled anti (antiligand) for the antiligand acted from the first group of compounds 8. Procedure according to claims 1 to 5, characterized in that the insolubilized representative is present in excess over the analyte and that both the solution of the analyte as well as the solution of the detectant are drawn into the syringe through the tip, to dissolve the analyte continuously to the unbound insolubilized representative suspend. 9. The method according to claims 1 to 8, characterized in that that the insoluble representative is an anti-ligand, which is present in excess compared to the analyte, and that the solution of the analyte removed from the tip, before introducing the detector, the tip with a Wash solution washes the detector solution through the syringe into the tip and through draws this through and removes the possibly existing non-specific bound detector washes the tip. 10. Procedure for Performing a Variety of Assay Determinations in a variety of samples, which is the analyte to be determined a representative of one of two families that have a pair of specifically bindable Substances comprise and wherein at least one sample contains a known amount of the analyte contains, in which one is insoluble for carrying out the assay determinations made representative of one of these families, a detective who is a Representatives of one of these families acts and that depending on the number of existing detectant molecules provides a physically measurable signal, and one Battery of syringes with plungers designed to achieve synchronous movement with each other tied together are attached and detachable on the ends of the syringes Tips for the uptake of the solution and the retention of the insolubilized Representative used, characterized in that one in each of the tips that the contain insoluble representatives, each draws up a different one of these samples, a detector solution from an external source or from syringes by means of these syringes or introduce any other reagent solutions and wash solutions into the tips, the solution is removed from the tips by means of the syringes and physically detectable signal is the amount of insolubilized detectant in the tips or the detectant removed from the tips as a measure of the amount of in each the analytes contained in the sample. 11. The method according to claim 10, characterized in that the makes insoluble representatives in the tip insoluble by means of the Syringe draws the insoluble representative in soluble form into the tip, whereby the insolubilized representative is bound to the surface of the tips will. 12. The method according to claim 10 or 11, characterized in that the insolubilized representative is used in an amount sufficient for binding of the total of the detectant and the total of the analyte is insufficient and that the Detectant marked radsuctively, this being made insoluble with respect to the Representatives can compete with the analyte. 13. The method according to claim 10 or 11, characterized in that the insoluble representative is in excess over the amount of what is present Analytes used that one radioactively labeled the detectant, the detectant and the insolubilized agent can bind the analyte. 14. The method according to claims 10 to 13, characterized in that that an antibody from a first group is used as an insoluble representative of compounds and, as a detectant, a combination of an antiligand of a second group of compounds and a radioactively labeled anti (antiligand) from this second group of compounds used. 15. The method according to claims 10 to 14, characterized in that that one uses as a representative made insoluble one that with the analyte can compete with respect to the detectant, and that the detectant is radioactive labeled, the detectant and the analyte with each other prior to introduction into the tips be combined. 16. The method according to claim 15, characterized in that as Analyte and, as the insoluble representative, representatives from the same family used, which are ligands selected from the Group of hold and antigens and that one as a detectant is a combination of one Antiligands and a radioactively labeled anti (anti-ligand) are used. 17. Device for performing a large number of simultaneous Transfers of liquids through the introduction and expulsion of solutions small tips that sit on the tip-receiving ends of syringes, g e k e n n -z e i c h n e t by a housing (12) with an upper platform (32) and a lower platform (50), one between the upper platform (32) and the lower platform (50) retained in a predetermined spatial relationship to one another A plurality of syringe barrels with tip receiving ends (86) extending from one another of the cylinders extending through the lower platform (50), a plurality of syringe plungers (76) disposed within the syringe barrel, wherein the pistons (76) have rods (100) extending beyond the upper platform (32) extend, means for simultaneously moving the pistons (76) in one vertical direction connected to an upper portion of the pistons (76), and means for removing the tips (90) from the syringe ends (86), the one between the lower platform (50) and the upper edge of the one on the syringe ends (86) seated tips (90) is mounted. 18. The device according to claim 17, characterized by a liquid reservoir or fluid reservoir (118), the fluid connection or fluid connection with openings stands, each of the cylinders having at least one opening in an upper one section the cylinder has. 19. Apparatus according to claim 17 or 18, characterized in that that the syringe plungers (76) are hollow on the inside. 20. Device according to claims 17 to 19, characterized in that that the means for removing the tips (90) from a plate (96) with a A plurality of openings through which the syringe ends (86) extend, one Cam means (106) operatively connected to the plate (96) for controlling the Plate (96) lowering to push the tips (90) out, and an elastic one There is means for returning the plate (96) to its rest position. 21. Device for the recording and control of the device according to Claims 17 to 20, characterized by a frame (16), a holding device for rigidly holding the piston moving device in an upper section of the frame (16), a carrier and locking device for holding the lower Platform (50) in a stationary position in a lower portion of the frame (16), a lifting and lowering device that is operable with the piston holder device is connected to raise and lower the pistons (76) to predetermined positions and driving means connected to the raising and lowering means is. 22. The device according to claim 21, characterized by a carrier platform (40) below the holding device for the lower platform (50). 23. The device according to claim 22, characterized by a lifting and lowering means operatively connected to the support platform (40). 24. Device according to claims 21 to 23, characterized in that that the raising and lowering device for the piston moving device a plurality of threaded rods (100) which form the support means for the piston movement device hold by means of a screw connection, and gears (30), which is operatively connected to the drive device in drive connection with the threaded support rods (100) are connected. 25. Device according to claims 21 to 24, characterized in that that the raising and lowering means for the support means one hydraulic device is. 26. Device according to claims 21 to 25, characterized in that that the programmed control device is operatively connected to the drive device is. 27. Small conical tip for use in immunoassays and For attachment to syringe ends, characterized by a large number of spacer arms, which are bound to the inner surface of the tip and are covalent or non-covolent binding can form proteins and polypeptides. 28. Conical tip according to claim 27, characterized in that the spacer arms are anti-ligands. 29. Conical tip according to claim 27, characterized in that the spacer arms have functional groups that correspond to amino groups of the proteins and polypeptides can react to form amide or amidine bridges. 30. Conical tip according to claims 27 to 30, characterized in that that they have a lower opening with a diameter within the range of 0.25 to 2 mm. 31. Conical tip according to claims 27 to 30, characterized in that that it has an uneven inner surface.