KR20050020985A - Determining the interaction between a C-reactive protein and constituents that bind to a C-reactive protein - Google Patents

Abstract

The present invention provides a method for determining the concentration of a solution containing CRP or C1q and for identifying substances that affect the interaction between CRP or C1q and the components that bind to them, in particular the interaction between CRP and C1q. ) Or an HTS-enabled method and test system for measuring the interaction between C1q and a component that binds CRP or C1q.

Description

Determining the interaction between a C-reactive protein and constituents that bind to a C-reactive protein}

The present invention provides an HTS-capable method and assay system for measuring the interaction between C-reactive protein (CRP) and C1q and components that bind CRP and C1q, respectively, for determining the concentration of CRP and C1q and for determining the concentration of CRP and C1q. And the determination of substances that affect the interaction between each of C1q and the components that bind to them.

C-reactive protein (CRP) is an acute plasma protein whose serum concentration rapidly increases significantly after infection or tissue injury (Volanakis (2001), Molecular Immunology 38, 189-197). CRP binds to phosphocholine (PCh) in the presence of Ca ²⁺ ions. Phosphocholine is very common in the polysaccharides of pathogenic organisms and the cell membranes of damaged and necrotic cells. CRP bound to PCh can activate the classical complement cascade by binding to protein C1q.

Complement is part of the immune system and is primarily involved in antibody-mediated immune defense. The three physiological functions of complement are defense against bacterial infections, the linkage of innate and acquired immunity and the elimination of immune complexes and apoptotic cells. There is a difference between the classic complement multistep, the alternative complement multistep and the mannose-lectin complement multistep (Walport (2001), N Engl J Med 344, 1958-1066). The classical complement multistep is initiated with C1q or PCh-bound CRP that induces lysis of bacterial cells and associates with antibodies that bind to the cell surface.

C-reactive protein is used as a marker and also as a predictor for coronary heart disease (Rifai and Ridker (2001), Clinical Chemistry 47, 403-411), the most common cause of death in developed countries.

Due to a new study (Jialal et al (2001), Circulaton 103, 1993-1935), lowering CRP levels or blocking complement activation by binding to CRP-mediated effector functions, eg, C1q. It may be useful for the prevention and treatment of coronary heart disease.

Therefore, it is a first priority to provide a method which enables the measurement of the concentration of CRP and / or C1q in plasma in a simple manner, and secondly, the interaction between CRP and / or C1q and other components, in particular between CRP and C1q. It is desirable to provide a method that enables the identification of substances acting on an interaction in a controlled, ie activated or inhibited manner.

Methods for measuring the binding of CRP to C1q are described, for example, in Jiang et al. (1991), J. Immunol. 146, 2324-2330 and Agrawal et al. (2001), J. Immunol. 166, 3998-4004. Both of these documents describe the use of ELISA methods. Both of these documents are very time consuming and material consuming multistage non-uniform methods. Thus, to carry out the method, it is always necessary to first immobilize one of the reagents on the solid phase. In addition, multiple washing steps are required.

See, eg, Lebdue et al. (1998), Ann Clin Biochem 35, 745-753, describe a method for immunoturbidity detection of CRP. To this end, samples are incubated with anti-CRP antibodies loaded with polystyrene particles. This method has the critical disadvantage that a turbidimeter is required and that a relatively large amount of sample must be used to carry out the method.

Accordingly, it is an object of the present invention to allow the concentration of CRP or C1q in plasma to be measured in a simple manner and also to regulate and activate substances which act on the interaction between CRP or C1q and other components, in particular the interaction between C1q and Crp. Or a method that enables identification in a suppressive manner, which is advantageous over the prior art in that it is performed more quickly than simple and / or already described methods that are more sensitive, cost-saving and material-saving. Are distinguished.

The purpose is

(a) initially introducing a CRP containing or C1q containing solution,

(b) a component comprising one or more compounds or groups of compounds (donor groups) capable of emitting signals after being excited by a light source and capable of binding to CRP or C1q directly or through one or more components of said groups Adding at least one donor component or component group containing at least one donor component to the CRP containing or C1q containing solution,

(c) a component comprising at least one compound or group of compounds (a group of receptors) capable of receiving a signal emitted by the compound or group of compounds according to step (b) and releasing it in the form of electromagnetic radiation and directly or in the group Adding to the CRP containing or C1q containing solution one or more receptor components or groups of components containing one or more receptor components capable of binding to CRP or C1q via one or more components,

(d) exciting at least one compound or group of compounds (donor group) according to step (b) with a light source, and

(e) detecting electromagnetic radiation emitted by at least one compound or group of compounds (receptor group) according to step (c) to detect binding events between two components that bind CRP or C1q directly or through one or more components A method of determining the binding event between two components that bind CRP or C1q, either directly or through one or more components.

The emitted electromagnetic radiation according to step (c) is preferably fluorescent radiation. The light source according to step (d) can be, for example, a laser or a lamp, for example a helium or halogen lamp. The electromagnetic radiation according to step (e) can be detected for example with the aid of a photomultiplier.

The component used according to the invention is preferably a polypeptide or comprises a polypeptide. In a particularly preferred embodiment of the invention, at least one of the components is an antibody, which may be a monoclonal or polyclonal antibody.

When component groups are used, the components can preferably be bonded to CRP or C1q or to one another in space continuously. The binding occurs in a binding multistage form, for example, the continuous binding of an antibody (primary antibody, secondary antibody, etc.). Herein, the component directly binding to CRP or C1q according to steps (b) and (c) is preferably a component which in each case binds to a specific binding region or a specific epitope of CRP or C1q, wherein step (b The CRP-binding component or C1q-binding component according to) is bound to a different binding site than the CRP- or C1q-binding component according to step (c).

In another preferred embodiment of the invention, one of the components directly binding to CRP or C1q is the natural binding partner of CRP or C1q, which binding partner is particularly preferably C1q in the case of CRP and vice versa. If the donor or acceptor component binds directly to CRP or C1q, in this case the donor or acceptor component is C1q or CRP correspondingly comprising the donor or receptor group. When using component groups capable of forming binding multistages, C1q or CRP is the first component correspondingly to CRP or C1q, through which a donor or receptor component can bind to CRP or C1q. The remaining components of the component group are particularly preferably antibody molecules.

Thus, for example, when initially introducing a CRP containing solution, in this embodiment preferably the donor or acceptor component itself comprises C1q, or the donor or acceptor component is bound to CRP via C1q. In such cases, the donor or receptor component may comprise an anti-C1q antibody, or a secondary antibody capable of binding to such anti-C1q antibody. Thus, in each case the other component (receptor or donor component) comprises an anti-CRP antibody or an antibody capable of binding to such anti-CRP antibody or a tertiary antibody directed against said antibody.

In a preferred embodiment, the at least one compound or group of compounds according to step (b) (donor group) and / or step (c) (receptor group) has a particulate diameter of preferably from 150 to 250 nm, particularly preferably from about 200 nm. It can be located at Thus, the donor or acceptor component comprises the particles, in which case the particles can be made of polymeric material.

The method of the present invention is an HTS (High Throughput Screening) -capable method that can be performed uniformly in the form of a one step method.

In a preferred embodiment, the method of the present invention is carried out according to the "mix and measure" principle which leads to significant time savings and high accuracy of the measured data.

The method of the present invention makes it possible to determine the binding of CRP to C1q to be measured, to identify substances affecting the binding, and to determine the concentration of CRP or C1q in the composite medium, as described previously. When the method is used, the measurement can be made within 10% of the time required.

A particular characteristic of this new method is that it allows monitoring of the binding between two proteins in solution, since no reactants need to be immobilized on the solid phase. In addition, proteins obtained from biological sources can be measured directly, for example, without the need to bind fluorophores to modify CRP or C1q. Thus, the method has the advantage that the protein can be introduced into its assay in its original state, eliminating the risk of denaturation that may be present when the protein is fixed and / or modified.

Another advantage of the process of the invention is the sample volume of the process of the invention, which is considerably reduced compared to conventional methods. Thus, volumes less than 10 μl may be used to minimize material costs. In addition, highly diluted solutions with CRP or C1q concentrations below 1 nM, even 100 pM, can be used. In addition, the method of the present invention is a very sensitive method. In addition, due to the above-mentioned description, the method of the present invention is the first to enable high throughput screening of substances that affect complement activation due to the binding of CRP to C1q.

In a preferred embodiment of the invention, the signal is transferred via singlet oxygen to at least one compound or compound group (donor group) according to step (a) to at least one compound or compound group (receptor group) according to step (b) do.

In such a case, the donor group preferably comprises a compound capable of converting triplet oxygen to singlet oxygen after excitation by a laser, and the receptor group is one or more first compounds that can be excited by singlet oxygen and And a second compound capable of absorbing energy absorbed by the group which may be excited by singlet oxygen in an emissionless manner and releasing it in the form of fluorescent radiation.

The singlet oxygen formed can diffuse from the donor group to the receptor group and react with the chemiluminescent material present in the receptor group. The energy released in this process is transferred to a fluorophore which finally emits this energy in the form of fluorescent radiation which can be detected using a photomultiplier. Since singlet oxygen is unstable and disintegrates in aqueous solution, a prerequisite for the detectable signal is the spatial accessibility of the donor and acceptor beads. Thus, the binding component to be used in the method is chosen such that when a binding event occurs, the distance between the donor group and the acceptor group is preferably less than 200 nm, which is due to the singlet oxygen at larger distances due to the decay time of singlet oxygen. This is because the effective energy transfer is impossible.

In one embodiment, it is particularly preferred that the donor group and the acceptor group are located on particles preferably having an average diameter of 150 to 250 nm, particularly preferably about 200 nm. Preferred use of such particles herein is such that the final concentration of the donor group bearing particles is 1-40 μg / ml and the final concentration of the receptor group bearing particles is 1-80 μg / ml. The binding performance of the particles can be, for example, about 0.1 nM to 1 nM per μg / ml of particles.

Particular preference is given to using alpha-screen-beads (Perkin-Elmer Life Sciences) as particles in the process. In this embodiment, the donor group can be excited by radiation irradiation at a wavelength of 680 nm using a laser and the radiation emitted by the receptor group can be detected at a wavelength of 520-600 nm.

Thus, in this case the method can be carried out such that donor and acceptor beads bearing the donor and acceptor groups are initially introduced. Subsequently, a component capable of binding CRP or C1q either directly or through additional components is bound to the donor and acceptor beads. As already mentioned above, these components may be C1q or CRP or may be anti-CRP or anti-C1q antibodies, or may be secondary antibodies directed against the anti-CRP or anti-C1q antibodies.

In a further preferred embodiment of the method of the invention, the signal is obtained from step (b) from one or more compounds or compound groups (donor groups) according to step (b) via emission-free energy transfer, particularly preferably via fluorescence resonance energy transfer. To one or more compounds or groups of compounds (receptor groups).

In this case, for example, at least one compound or compound group (donor group) according to step (a) comprises a europium salt containing compound and at least one compound or compound group (receptor group) according to step (b) Methods can be performed to include phycocyanin (Grepin et al. (2000), Drug Discovery Today 5, 212). Alternatively, the donor group and the acceptor group may be dyes suitable for no emission energy transfer. In addition, all compounds known to those skilled in the art, suitable for emission-free energy transfer, in particular fluorescence resonance energy transfer, can be used. Pope et al. (1999), Drug Discovery Today 4, 350].

In this embodiment, it is preferable to select the binding component to be used such that the distance between the donor group and the acceptor group after binding is less than 10 nm, since effective no-emission energy transfer is impossible at larger distances.

In a preferred embodiment according to the invention, the method of the invention is used to determine the concentration of a CRP containing or C1q containing solution of unknown CRP or C1q content, which solution is preferably plasma or blood serum or suitable physiological Blood plasma or blood serum diluted with buffer or water. The method can be used, for example, for diagnostic purposes, in particular for measuring the inflammatory state of an organism and / or for measuring the risk of cardiac arrest and / or heart attack.

Accordingly, the present invention relates to an HTS-capable diagnostic assay system for measuring the binding event between CRP or C1q and a component that binds to CRP or C1q comprising: (a) a signal after being excited by a light source A component comprising one or more compounds or groups of compounds (donor groups) capable of releasing and containing one or more donor components or one or more donor components capable of binding to CRP or C1q directly or through one or more components of said groups Ingredient group and

(b) a component comprising at least one compound or group of compounds capable of receiving a signal emitted by the compound or group of compounds according to (a) and capable of releasing in the form of electromagnetic radiation and directly or at least one component of said group of components A component group containing one or more receptor components or one or more receptor components capable of binding to CRP or C1q via.

The HTS-enabled assay system herein comprises the following additional components:

(c) blood serum or blood plasma, or blood diluted with physiological buffer or water,

(d) a light source for exciting one or more compounds or group of compounds according to (a) and

(e) a detection system for detecting the emitted electromagnetic radiation according to (b).

In another preferred embodiment of the method of the invention, the at least one test substance is prepared before step (a) and / or before step (b) and / or before step (c) to observe whether such test substance affects the binding event. And / or before step (d). In this way, substances that act on the interaction between CRP or C1q and the binder, in particular the interaction between CRP and C1q, can be measured in a modulated, inhibited or activated manner. This involves screening libraries of test materials, preferably by HTS methods, to determine materials with desired properties.

Accordingly, the present invention relates to an HTS-capable assay system for determining, in a modulated, inhibited or activated manner, a substance that acts on the interaction between CRP or C1q and a component that binds to CRP or C1q, comprising:

(a) a component comprising one or more compounds or groups of compounds (donor groups) capable of emitting signals after being excited by a light source and capable of binding to CRP or C1q directly or through one or more components of said groups A component group containing at least one donor component or at least one donor component,

(b) a component comprising at least one compound or group of compounds capable of receiving a signal emitted by the compound or group of compounds according to (a) and capable of releasing in the form of electromagnetic radiation and directly or at least one component of said group of components A component group containing one or more receptor components or one or more receptor components capable of binding to CRP or C1q via

(c) one or more test compounds.

The HTS-enabled assay system herein preferably comprises the following additional components:

(d) a light source for exciting one or more compounds or group of compounds according to (a),

(e) a detection system for detecting the emitted electromagnetic radiation according to (b) and

(f) CRP containing or C1q containing solutions.

The HTS-enabled assay system of the present invention is preferably used to carry out the method of the present invention described above. Thus, certain aspects of the portions and components of the assay system correspond to the aforementioned specific aspects used in the methods of the invention.

Scheme 1 is a schematic illustration of the method described in the exemplary embodiment. In the above scheme, the donor component used is an anti-C1q antibody and the donor group is located on particles bound to the anti-C1q antibody. In this case, the donor component may bind to CRP via C1q. The receptor component in this case is an anti-rabbit secondary antibody and the receptor group is located on a particle bound to the anti-rabbit secondary antibody. In this case, the receptor component can bind CRP via an anti-CRP antibody derived from rabbits.

Scheme 1

Exemplary Embodiments: Binding Assay to Measure Binding of CRP to C1q

Experiments have shown that a donor bead comprising a compound capable of converting triplet oxygen to singlet oxygen after being excited at a defined wavelength, and a compound that can be excited by singlet oxygen and energy from the excited compound After receiving in metastatic form, it was performed using an Alphascreen Detection Kit (Packard Bioscience) containing receptor beads containing compounds capable of releasing in the form of fluorescent radiation. The compounds mentioned are in each case embedded in the hydrogel matrix. The beads used were precoated by the manufacturer, ie the donor beads were streptavidin and the receptor beads were precoated with anti-rabbit antibody.

Overview of the materials used Reagent / Plate Molecular weight or concentration Source, catalog number Fine Plate, 384K, PS, White Greiner 784075 C-reactive protein, human, tooth. Recombinant from E. coli 1 mg / ml, 115 kd (pentamer) Calbiochem 236608 c1q, person 1mg / ml, 410kd Calbiochem 204876 Anti-CRP, Rabbit 3.57mg / ml, 165kd Calbiochem 235752 Alpha Screen IgG Detection Kit (Protein A) Packard BioScience 6760617 Alpha Screen Rabbit IgG Detection Kit Packard BioScience 6760607 Anti-c1q, chlorine Polyclonal serum Calbiochem 234390 Anti-streptavidin Polyclonal serum Sigma S6390 EZ-Linked Sulfo-NHS-LC-Biotinylation Kit Pierce 21430 Sodium chloride 58.44 g / mol Merck 101540 Potassium Chloride (CaCl ₂ × 2H ₂ O) 147.02 g / mol Merck 102382 Tris (hydroxymethyl) -aminomethane hydrochloride (tris) 157.6 g / mol Merck 108219 Bovine Serum Albumin (BSA) Sigma P7888 Phosphoryl choline chloride 329.7 g / mol Sigma P0378 Phosphate Buffered Saline without Mg ²⁺ and Ca ²⁺ (PBS) Gibco BRL14200-067 Slide-A-Laser Mini Dialysis Unit, 10,000 MWCO Pierce P.69570 Dimethyl Sulfoxide (DMSO) Merck KGaA, 1.02931 ethanol Riedel de Haen, 32250 Pluronic F-68 10% strength solution Sigma, P5556 Hydrochloric acid 1M Merck, 109057 Sodium hydroxide solution 1M Merck, 109 137 Milli-QH ₂ O

Preparation of Reaction Buffer

The following reaction buffer was used in the alpha-screen-system: 20 mM Tris-HCl, pH 7.2, 150 mM NaCl, 5 mM CaCl ₂ , 1 mM phosphoryl choline, 0.1% BSA. A mother liquor of Tris-NaCl buffer was prepared by dissolving 3.15 g of Tris-HCl and 8.77 g of NaCl in H ₂ O, adjusting the pH to 7.2 with 1 M NaOH, and then adding H ₂ O to 1 L. Buffer was stored at room temperature. Reaction buffer was prepared by adding 36.7 mg CaCl ₂ , 12.9 mg phosphoryl choline and 50 mg BSA to Tris-NaCl buffer.

Biotinylation of Anti-C1q Antibodies

In order to bind the anti-C1q antibody to streptavidin-coated donor beads, it was necessary to first biotinylate the anti-C1q antibody. To this end, 100 μl of anti-C1q polyclonal serum was dialyzed twice against 200 ml of PBS for 1 hour at room temperature. After addition of 20 μl of a 1.5 mM sulfo-NHS solution (in milli-Q water), the solution was left at room temperature for 30 minutes and then dialyzed twice against PBS to remove excess biotinylation reagent.

Black Course

The assay was initially performed by introducing donor beads / anti-CRP solution followed by addition of 2 μl C1q, 2 μl CRP and 2 μl of receptor beads / anti-CRP. This resulted in the following final concentrations: CRP: 1 nM; C1q: 10 nM; Anti-CRP: 7.3 nM; Anti-C1q: 1: 1500; Donor beads: 20 μg / ml: Receptor beads: 40 μg / ml. As a negative control, CRP solution, C1q solution or both solutions were replaced with reaction buffer. After incubation for 2 hours at room temperature, the data were read with AlphaQuest reader.

result

The intensity of radiation emitted at a CRP concentration of 1 nM and a C1q concentration of 10 nM was measured as follows (describing the photomultiplier coefficients):

In the absence of CRP and in the absence of C1q: 952

In the presence of CPR and in the absence of C1q: 1255

In the absence of CRP and in the presence of C1q: 1114

In the presence of CRP and in the presence of C1q: 80376

As can be seen, the negative control does not give a positive result and strong signals are visualized when the C1q containing solution and CRP containing solution are combined.

Claims (36)

(a) initially introducing a C-reactive protein (CRP) containing solution, (b) one or more components comprising one or more compounds or compound groups (donor groups) capable of emitting signals after being excited by a light source and one or more components capable of binding to CRP directly or through one or more components of the component groups Adding a donor component or component group containing one or more donor components to the CRP containing solution, (c) a component comprising at least one compound or group of compounds (receptor group) capable of receiving a signal emitted by the compound or group of compounds according to step (b) and releasing it in the form of electromagnetic radiation; Adding to the CRP containing solution at least one receptor component or a group of components containing at least one receptor component capable of binding to the CRP through at least one component of (d) exciting at least one compound or group of compounds (donor group) according to step (b) with a light source, and (e) detecting electromagnetic radiation emitted by at least one compound or group of compounds (receptor group) according to step (c) to detect a binding event between two components that binds to the CRP either directly or through one or more components A method for measuring a binding event between two components, comprising, directly or through one or more components to the CRP. (a) initially introducing a C1q containing solution, (b) one or more components or compounds comprising one or more compounds or groups of compounds (donor groups) capable of emitting signals after being excited by a light source and capable of binding to C1q directly or through one or more components of those component groups Adding a donor component or a component group comprising one or more donor components to the C1q containing solution, (c) a component comprising at least one compound or group of compounds (receptor group) capable of receiving a signal emitted by the compound or group of compounds according to step (b) and releasing it in the form of electromagnetic radiation; Adding one or more receptor components or a group of components containing one or more receptor components capable of binding to C1q through one or more components of to the C1q containing solution, (d) exciting at least one compound or group of compounds (donor group) according to step (b) with a light source, and (e) detecting electromagnetic radiation emitted by at least one compound or group of compounds (receptor group) according to step (c) to detect a binding event between two components that bind C1q directly or through one or more components Method for measuring the binding event between two components that bind to C1q directly or through one or more components comprising. The method according to claim 1 or 2, wherein the electromagnetic radiation is fluorescent radiation. The method according to claim 1 or 2, wherein the light source according to step (d) is a laser or a lamp. The method according to claim 1 or 2, wherein the detection according to step (e) is carried out with the aid of a photomultiplier. The method according to claim 1, wherein the components of the component group according to step (b) and / or step (c) can be bonded to the CRP or to each other continuously in space. The method of claim 2, wherein the components of the component group according to step (b) and / or step (c) can bind to C1q or to each other continuously in space. 8. The method of any one of claims 1 to 7, wherein the component is or comprises a polypeptide. The method of claim 8, wherein at least one of the components is an antibody. The method of claim 9, wherein the antibody is a monoclonal or polyclonal antibody. The method of claim 1, wherein at least one of the components is a natural binding partner of the CRP. The method of claim 11, wherein the natural binding partner of CRP is C1q. The method of claim 12, wherein the component group according to step (a) of claim 1 or step (b) of claim 1 comprises C1q and anti-C1q antibodies. The method of claim 2, wherein at least one of the components is a natural binding partner of C1q. The method of claim 14, wherein the natural binding partner is CRP. The method of claim 15, wherein the component group according to step (a) of claim 2 or step (b) of claim 2 comprises CRP and anti-CRP antibodies. The compound according to claim 1 or 2, wherein the signal is one or more compounds or compounds according to step (c) from one or more compounds or compound groups (donor groups) according to step (b) by means of emissionless energy transfer. How to metastasize to a group (receptor group). 18. The method of claim 17, wherein the emission free energy transfer is a fluorescence resonance energy transfer. The method of claim 1 or 2, wherein the signal is transferred via singlet oxygen. 20. The method of claim 19, wherein the triplet oxygen is converted to singlet oxygen after the at least one compound or group of compounds (donor group) according to step (b) or step (b) of claim 1 is excited by a laser. At least one first compound comprising a compound which can be excited and wherein at least one compound or group of compounds (receptor group) according to step (b) of claim 1 or step (b) of claim 2 can be excited by singlet oxygen. And at least one second compound that is capable of absorbing energy absorbed by the first compound that can be excited by singlet oxygen in a non-emission manner and releasing it in the form of fluorescent radiation. The method of claim 20, wherein the donor group and / or the receptor group are located on the particle. The method of claim 21, wherein the average diameter of the particles is about 200 nm. The method of claim 1, wherein the CRP content is used to determine the concentration of the unknown CRP containing solution. 23. The method of any one of claims 1 to 22, wherein the C1q content is used to determine the concentration of the unknown C1q containing solution. The method of claim 23 or 24, wherein the solution is blood serum, blood plasma, or blood serum or blood plasma diluted with physiological buffer or water. The method of claim 25, which is a diagnostic method for measuring the inflammatory state of an organism and / or for determining the risk of cardiac arrest and / or heart attack. 27. The method of any one of claims 1 to 26, which is an HTS-capable uniform method that can be performed as a one step method. The method according to claim 1, wherein at least one test substance is added before step (a) and / or before step (b) and / or before step (c) and / or before step (d) such that the test substance is selected from CRP and CRP-. And a further step of observing whether the binding event between the binding components is affected. The method according to claim 2, wherein at least one test substance is added before step (a) and / or before step (b) and / or before step (c) and / or before step (d) so that the test substance is selected from C1q and C1q-. And a further step of observing whether the binding event between the binding components is affected. The method of claim 28 or 29, which is used to measure a substance acting on a binding event in a modulated, inhibited or activated manner. (a) one or more components or compounds comprising a compound group (donor group) capable of emitting a signal after being excited by a light source and which can bind to the CRP directly or through one or more components of the component group A component group comprising a donor component or one or more donor components, and (b) a component comprising at least one compound or group of compounds (receptor group) capable of receiving a signal emitted by the compound or group of compounds according to (a) and releasing it in the form of electromagnetic radiation and directly or of the group of components An HTS-capable diagnostic assay system for measuring the binding event between a CRP and a component that binds to CRP, comprising at least one receptor component capable of binding to the CRP through one or more components or a group of components containing one or more receptor components. (a) one or more compounds or compounds comprising a compound group (donor group) capable of emitting a signal after being excited by a light source and which can bind to C1q directly or through one or more components of that component group A component group comprising a donor component or one or more donor components, and (b) a component comprising at least one compound or group of compounds (receptor group) capable of receiving a signal emitted by the compound or group of compounds according to (a) and releasing it in the form of electromagnetic radiation and directly or of the group of components An HTS-capable diagnostic assay system for measuring the binding event between C1q and a component that binds to C1q, comprising one or more receptor components capable of binding to C1q through one or more components or a component group containing one or more receptor components. 33. The method of claim 31 or 32, (c) blood serum or blood plasma, or blood serum or blood plasma diluted with physiological buffer or water, (d) a light source capable of exciting one or more compounds or compound groups (donor groups) according to (a) and (e) an HTS-enabled assay system comprising additional components of a detection system for detecting emitted electromagnetic radiation according to (b). (a) one or more components or compounds comprising a compound group (donor group) capable of emitting a signal after being excited by a light source and which can bind to the CRP directly or through one or more components of the component group A component group comprising a donor component or one or more donor components, (b) a component comprising at least one compound or group of compounds (receptor group) capable of receiving a signal emitted by the compound or group of compounds according to (a) and releasing it in the form of electromagnetic radiation and directly or of the group of components One or more receptor components or groups of components containing one or more receptor components capable of binding to the CRP through one or more components, and (c) HTS-capable assay system for controlling, in a controlled manner, an active substance that acts on the interaction between CRP and a component that binds to CRP, comprising one or more test compounds. (a) one or more compounds or compounds comprising a compound group (donor group) capable of emitting a signal after being excited by a light source and which can bind to C1q directly or through one or more components of that component group A component group comprising a donor component or one or more donor components, (b) a component comprising at least one compound or group of compounds (receptor group) capable of receiving a signal emitted by the compound or group of compounds according to (a) and releasing it in the form of electromagnetic radiation and directly or of the group of components One or more receptor components or groups of components containing one or more receptor components capable of binding to C1q through one or more components, and (c) HTS-enabled assay system for determining the active substance that acts on the interaction between C1q and a component that binds C1q, comprising one or more test compounds. The method of claim 34 or 35, (a) a light source capable of exciting one or more compounds or compound groups (donor groups) according to (a) and (b) an HTS-enabled assay system comprising additional components of a detection system for detecting emitted electromagnetic radiation according to (b).