JP2005529348A - HTS capable method and test system for measuring the interaction between C-reactive protein and components binding to C-reactive protein - Google Patents

Abstract

The present invention is for measuring the concentration of a solution containing C-reactive protein (CRP) or C1q, and the interaction of CRP or C1q with components that bind to them, in particular the interaction between CRP and C1q. The invention relates to HTS capable methods and test systems for measuring the interaction between CRP or C1q and components that bind to CRP or C1q in order to identify substances that affect the system.

Description

  The present invention measures the concentration of a solution containing C-reactive protein (CRP) and C1q, respectively, and measures substances that affect the interaction between CRP and C1q and the components that bind to them, respectively. Therefore, it relates to an HTS capable method and analysis system for measuring the interaction of CRP and C1q with components that bind to CRP and C1q, respectively.

C-reactive protein (CRP) is an acute phase plasma protein whose serum concentration increases rapidly and significantly after infection or tissue damage (Volanakis (2001), Molecular Immunology 38, 189-197). CRP binds to phosphocholine (PCh) in the presence of Ca ²⁺ ions. PCh is extremely ubiquitous in polysaccharides of pathogenic microorganisms and in 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 a component of the immune system and is primarily involved in immune defense mediated by antibodies. The three physiological functions of complement are defense against bacterial infection, the link between innate and acquired immunity, and the removal of immune complexes and apoptotic cells. The classical, alternative and mannose-lectin complement cascades are distinguished (Walport (2001), N Engl J Med 344, 1058-1066). The classical complement cascade lyses bacterial cells and first initiates ligation of C1q with an antibody or PCh-binding CRP that binds to the cell surface.

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

  According to new studies (Jialal et al. (2001), Circulation 103, 1933-1935), CRP levels are reduced, or CRP-mediated effector functions (eg complement activation by binding to C1q) It is presumed that blocking is useful for the prevention and treatment of coronary heart disease.

  Therefore, the plasma concentration of CRP and / or C1q is first measured in a simple manner and then adjusted to the interaction between CRP and / or C1q and other components, in particular the interaction between CRP and C1q. It would be desirable to provide a method that can identify substances that act in a manner (ie, activate or inhibit).

  A method for measuring the binding between CRP and C1q is described in, for example, Jiang et al. (1991), J. MoI. Immunol. 146, 2324-2330, and Agrawal et al. (2001), J. MoI. Immunol. 166, 3998-4004. All documents utilize the ELISA method. Both of these are multi-stage, non-uniform methods that are extremely time and material consuming. Therefore, in order to carry out these methods, one of the reactants must always be fixed in advance on the solid phase. In addition, a number of cleaning steps are required.

  Lebdue et al. (1998), Ann Clin Biochem 35, 745-753, describe an immunoturbidimetric detection method for CRP. For this purpose, the sample is incubated with an anti-CRP antibody loaded on polystyrene particles. The method has significant disadvantages in that a nephelometer is required and that a relatively large amount of sample must be used to perform the method.

  Therefore, the object of the present invention is to measure the plasma concentration of CRP or C1q in a simple manner, and also to regulate, activate or interact with the interaction of CRP or C1q with other components, in particular with C1q and CRP. It is to provide a method that can identify substances that act in an inhibitory manner, said method being more sensitive, cost effective and saving material compared to previously described methods. It is advantageous over the prior art in that it is more convenient and / or faster.

The above objective is accomplished by a method for measuring the binding phenomenon of two components that bind to CRP or C1q, either directly or via one or more components, the method comprising the following steps:
(A) first introducing a solution containing CRP or C1q;
(B) adding at least one donor component or a component group containing at least one donor component to the CRP- or C1q-containing solution [the donor component emits a signal after being excited by a light source; A component comprising at least one compound or compound group (donor group) capable of binding to CRP or C1q directly or via one or more components from said component group can do],
(C) adding at least one acceptor component or a component group containing at least one acceptor component to the CRP- or C1q-containing solution [wherein the acceptor component is a compound or a compound group described in (b) A component comprising at least one compound or group of compounds (acceptor group) capable of receiving the emitted signal in the form of receiving and electromagnetic radiation, said acceptor component being directly or of said group of components Can bind to CRP or C1q through one or more components]
(D) exciting at least one compound or compound group (donor group) according to (b) with a light source;
(E) detecting electromagnetic radiation emitted by at least one compound or group of compounds (acceptor group) according to (c) in order to detect the binding phenomenon.

  In the present invention, the emitted electromagnetic radiation described in (c) is preferably fluorescent radiation. The light source described in (d) may be, for example, a laser or a lamp, such as a helium or halogen lamp. The electromagnetic radiation described in (e) may be detected using, for example, a photomultiplier tube.

  Preferably, the component that can be used in the present invention is a polypeptide or comprises a polypeptide. In a particularly preferred embodiment of the invention, at least one said component is an antibody, which may be a monoclonal antibody or a polyclonal antibody.

When a component group is used, the components can preferably be bound spatially continuously, to CRP or C1q, or to each other. The binding occurs in the form of a binding cascade, examples of which include continuous binding of antibodies (primary antibodies, secondary antibodies, etc.). In the present invention, the component that directly binds to CRP or C1q described in (b) and (c) preferably binds to a specific binding region or a specific epitope of CRP or C1q in any case. And the CRP or C1q binding component described in (b) binds to a different binding site than the CRP or C1q binding component described in (c).

  In other preferred embodiments of the method of the invention, one of the components that binds directly to CRP or C1q is a naturally occurring binding partner of CRP or C1q, and in the case of CRP, such a partner is particularly preferably C1q. And vice versa. Thus, when the donor or acceptor component binds directly to CRP or C1q, either the donor or acceptor component is either a C1q or CRP containing donor group or acceptor group. Thus, when using a group of components capable of forming a binding cascade, C1q or CRP is the first component that binds to CRP or C1q, through which either the donor or acceptor component is transferred to CRP. Alternatively, it can be bound to C1q. The remaining component of the component group is particularly preferably an antibody molecule.

  Thus, for example, when initially introducing a CRP-containing solution, preferably in this embodiment either the donor or acceptor component itself contains C1q, or the donor or acceptor component is mediated via C1q. Bind to CRP. In this case, the donor or acceptor component may comprise an anti-C1q antibody or a secondary antibody that can bind to the anti-C1q antibody. Thus, in any case, the other component (acceptor or donor component) comprises an anti-CRP antibody, or an antibody capable of binding to said anti-CRP antibody, or a tertiary antibody directed against the latter. .

  In a preferred embodiment, at least one compound or compound group (donor group) described in (b) and / or at least one compound or compound group (acceptor group) described in (c) is The average diameter of the particles is preferably 150 to 250 nm, particularly preferably about 200 nm. Accordingly, in this case, the donor or acceptor component includes the particles, and the particles can be made of a polymer material.

  The method of the present invention is a method capable of high-throughput screening (HTS), and the method can be uniformly performed in the form of a one-step method.

In a preferred embodiment, the method of the present invention comprises a “mix and measure (mix
and measure) ”principle, which can save time and provide highly accurate measurement data.

  By the method of the present invention, the binding of CRP to C1q can be measured, so that in less than 10% of the time required when using the previously described heterogeneous method, Substances that affect binding and the concentration of CRP or C1q in the complex medium to be measured can be identified.

  A notable feature of this new method is that it is possible to monitor the binding between two proteins in solution since the reactants need not be immobilized on a solid phase. Moreover, proteins obtained from biological sources can be measured directly, and there is no need to modify CRP or C1q, for example by attaching fluorophores. Therefore, the method has the particular advantage that the protein can be analyzed in its natural state, thus eliminating the risk of denaturation that can occur when the protein is immobilized and / or modified. .

Another advantage of the method of the present invention is that the sample volume of the method of the present invention can be significantly reduced compared to conventional methods. Thus, an amount of less than 10 μl can be used, thus minimizing material costs. Moreover, highly diluted solutions with CRP or C1q concentrations of less than 1 nM and even less than 100 pM can be used. In addition, the method is extremely sensitive. Moreover, according to the above observations, this is the first method that has enabled high-throughput screening of substances that affect complement activation by binding CRP to C1q.

  In a preferred embodiment of the method of the invention, the signal is transmitted from at least one compound or compound group (donor group) according to (b) to at least one compound or compound group (acceptor according to (c). Group) via singlet oxygen.

  In this case, the donor group preferably comprises a compound capable of converting triplet oxygen to singlet oxygen after excitation by a laser, and the acceptor group comprises at least one first species that can be excited by singlet oxygen. And a second compound that can absorb energy absorbed by the group excitable by singlet oxygen in an emission-less manner and release it in the form of fluorescent radiation.

  The formed singlet oxygen can diffuse from the donor group to the acceptor group and react with the chemiluminescent substance present there. The energy released in this process is transferred to the fluorophore and finally releases the energy in the form of fluorescent radiation, which can be detected using a photomultiplier tube. A prerequisite for a detectable signal is that in aqueous solution, singlet oxygen is unstable and decays so that the donor and acceptor beads are in spatial proximity. Therefore, the binding component that can be used in this method is selected such that the distance between the donor group and the acceptor group is preferably less than 200 nm when the binding phenomenon occurs, because for longer distances, a single This is because effective energy transfer by singlet oxygen is impossible due to the decay time of the term oxygen.

  In this embodiment, particularly preferably, the donor group and the acceptor group are localized on the particles, and the average diameter of the particles is preferably 150 to 250 nm, particularly preferably about 200 nm. In this case, the preferred use form of the particles is that the final concentration of the donor group-containing particles is 1 to 40 μg / ml and the final concentration of the acceptor group-containing particles is 1 to 80 μg / ml. The binding capacity of the particles can be, for example, about 0.1 nM to 1 nM per 1 μg / ml of particles.

  Particularly preferably, alpha-screen beads from Perkin-Elmer Life Sciences are used as particles in the method. In this embodiment, the donor group is excited by emitting a laser at a wavelength of 680 nm, and the radiation emitted by the acceptor group is detectable at a wavelength of 520-600 nm.

  Therefore, in this case, the present method can be carried out in such a manner that donor beads and acceptor beads containing a donor group and an acceptor group are first introduced. Next, components that can bind to CRP or C1q either by themselves or through additional components bind to the donor and acceptor beads. As already mentioned above, these components can be either C1q or CRP, or an anti-CRP or anti-C1q antibody, or a secondary antibody directed against said anti-CRP or anti-C1q antibody.

In a further preferred embodiment of the method of the invention, the signal is derived from at least one compound or group of compounds (donor group) as described in (b) to at least one compound or group of compounds as described in (c) ( To the acceptor group) via emissionless energy transfer, particularly preferably via fluorescence resonance energy transfer.

  In this case, the method can be carried out as follows; for example, at least one compound or group of compounds (donor group) described in (b) comprises a europium salt-containing compound and (c ) May contain allophycocyanin (Grepin et al. (2000), Drug Discovery Today 5,212). Alternatively, the donor group and the acceptor group may be dyes suitable for emissionless energy transfer. In addition, all compounds known to those skilled in the art suitable for emissionless energy transfer, in particular fluorescence resonance energy transfer, can be used (see, eg, Pope et al. (1999), Drug Discovery Today 4,350).

  In this embodiment, it is preferable to select the binding component that is used so that the distance between the donor and acceptor groups after binding is less than 10 nm, because at longer distances the effective emission This is because less energy transmission is impossible.

  In a preferred embodiment according to the invention, the method of the invention is used to determine the concentration of a CRP or C1q containing solution with unknown CRP or C1q content, said solution preferably being plasma or serum, or Plasma or serum diluted with appropriate physiological buffer or water. The method can be used, for example, for diagnostic purposes, in particular to measure the state of inflammation of an organism and / or the risk of cardiac arrest and / or stroke.

Therefore, the present invention also relates to an HTS capable diagnostic analysis system for measuring the binding phenomenon between CRP or C1q and a component that binds to CRP or C1q, the system comprising the following components:
(A) at least one donor component or a component group comprising at least one donor component [the donor component is at least one compound or compound group capable of emitting a signal after being excited by a light source ( A donor group), which can bind to CRP or C1q directly or via one or more components from said group of components],
(B) at least one acceptor component or a component group comprising at least one acceptor component [wherein the acceptor component receives a signal emitted by the compound or the compound group described in (A) and forms of electromagnetic radiation A component comprising at least one compound or group of compounds that can be released at the same time, and the acceptor component is directly or via one or more components of the group of components to CRP or C1q Can be combined].

In this case, the HTS-capable analysis system preferably includes the following additional components:
(C) serum or plasma, or blood diluted with physiological buffer or water,
(D) a light source for exciting at least one compound or group of compounds according to (A),
(E) A detection system for detecting the emitted electromagnetic radiation according to (B).

  In other preferred embodiments of the method of the invention, in order to observe whether the test substance affects the binding phenomenon, step (a) and / or step (b) and / or step (c) and At least one test substance is added before / or step (d). In this method, it is possible to measure substances that act in a manner that regulates, inhibits or activates the interaction between CRP or C1q and the binder, in particular the interaction between CRP and C1q. This embodiment preferably includes screening a library of test substances with the HTS method to determine substances having the desired properties.

Therefore, the present invention also relates to an HTS-capable analytical system for measuring active substances acting in a manner that regulates the interaction between CRP or C1q and components that bind to CRP or C1q. Contains the following ingredients:
(A ′) at least one donor component or a component group including at least one donor component [the donor component is capable of emitting a signal after being excited by a light source; (Donor group), wherein the donor component can bind to CRP or C1q directly or via one or more components from the component group],
(B ′) at least one acceptor component or a group of components comprising at least one acceptor component [wherein the acceptor component receives and emits a signal emitted by the compound or group of compounds according to (A ′) and electromagnetic radiation A component comprising at least one compound or group of compounds that can be released in the form of: and the acceptor component is CRP or directly or via one or more components of the group of components Can bind to C1q],
(C ′) at least one test compound.

In this case, the HTS capable analytical system preferably comprises the following additional components:
(D ′) a light source for exciting at least one compound or group of compounds according to (A ′),
(E ′) a detection system for detecting emitted electromagnetic radiation according to (B ′),
(F ′) CRP or C1q-containing solution.

  In this case, the HTS capable analysis system of the present invention is preferably used to perform the method of the present invention described above. Therefore, specific embodiments of the elements and components of the analysis system correspond to the specific embodiments described above that are used in the method of the present invention.

  FIG. 1 schematically illustrates the method described in the examples. Here, the donor component used is an anti-C1q antibody, and the donor group is localized on the particle and binds to the anti-C1q antibody. In this case, the donor component can bind to CRP via C1q. In this case, the acceptor component is an anti-rabbit secondary antibody, and the acceptor group is localized on the particle and binds to the anti-rabbit secondary antibody. In this case, the acceptor component can bind to CRP via a rabbit-derived anti-CRP antibody.

Exemplary Embodiment: Binding Analysis to Measure CRP Binding to C1q This experiment was performed using an alpha-screen detection kit from Packard Biosciences, which includes donor and active Including a acceptor bead, wherein the donor bead includes a compound capable of converting triplet oxygen to singlet oxygen after being excited at a predetermined wavelength, and the acceptor bead is a compound capable of being excited by singlet oxygen; Further included are compounds capable of receiving energy from the excited compound in the form of emissionless energy transfer and then releasing it in the form of fluorescent radiation. Here the compounds were embedded in the hydrogel matrix in each case. The beads used were pre-coated by the manufacturer with donor beads being streptavidin and acceptor beads pre-coated with anti-rabbit antibodies.

Reaction Buffer Preparation The following reaction buffers were used in the alpha-screen system: 20 mM Tris-HCl (pH 7.2), 150 mM NaCl, 5 mM CaCl ₂ , 1 mM phosphorylcholine, 0.1% BSA. A stock solution of Tris-NaCl buffer was prepared by dissolving Tris-HCl (3.15 g) and NaCl (8.77 g) in H ₂ O, adjusting the pH to 7.2 with 1 M NaOH, and then H _2. Prepared by adding O to 1L. This buffer was stored at room temperature. Reaction buffer was prepared by adding CaCl ₂ (36.7 mg), phosphorylcholine (12.9 mg) and BSA (50 mg) to Tris-NaCl buffer (50 ml).

For biotinylated anti C1q antibody anti C1q antibody to be able to bind to streptavidin-coated donor beads,
The anti-C1q antibody must first be biotinylated. For this purpose, anti-C1q polyclonal serum (100 μl) was dialyzed twice against PBS (200 ml) for 1 hour at room temperature. After addition of 1.5 mM sulfo-NHS solution (MilliQ aqueous solution) (20 μl), this solution is allowed to stand for 30 minutes at room temperature and then twice against PBS to remove excess biotinylation reagent. Dialyzed.

Analytical Method Analysis was performed by first introducing donor beads / anti-CRP solution (2 μl) and then adding C1q (2 μl), CRP (2 μl) and acceptor beads / anti-CRP (2 μl). This gave the following final concentrations: CRP: 1 nM; C1q: 10 nM; anti-CRP: 7.3 nM; anti-C1q: 1: 1500; donor beads: 20 μg / ml; acceptor beads: 40 μg / ml. In the negative control, CRP solution, C1q solution, or both of these solutions were used instead of the reaction buffer. After incubating for 2 hours at room temperature, the data was read with an AlphaQuest reader.

Results The measured intensities of the radiation emitted at a CRP concentration of 1 nM and a C1q concentration of 10 nM were as follows (listing photomultiplier tube counts):
When CRP is absent and C1q is absent: 952
CRP present and C1q absent: 1255
CRP absent and C1q present: 1114
CRP present and C1q present: 80376.

  From the above, it can be seen that when the C1q-containing solution and the CRP-containing solution were combined, the negative control showed a strong signal but did not show a positive result.

Fig. 2 schematically shows the method described in the examples.

Claims (36)

A method for measuring the binding phenomenon of two components that bind to CRP directly or via one or more components, comprising the following steps:
(A) first introducing a CRP containing solution;
(B) adding at least one donor component or a component group containing at least one donor component to the CRP-containing solution [the donor component can emit a signal after being excited by a light source; A component comprising at least one compound or group of compounds (donor group), wherein the donor component can bind to CRP directly or via one or more components from the group of components ],
(C) adding at least one acceptor component or a component group containing at least one acceptor component to the CRP-containing solution [the acceptor component is released by the compound or compound group described in (b) A component comprising at least one compound or group of compounds (acceptor group) capable of receiving and emitting a signal in the form of receiving and electromagnetic radiation, wherein the acceptor component is directly or one or more of the component groups Can bind to CRP via further components],
(D) exciting at least one compound or compound group (donor group) according to (b) with a light source;
(E) detecting electromagnetic radiation emitted by at least one compound or group of compounds (acceptor group) according to (c) to detect the binding phenomenon;
Including the above method. A method for measuring the binding phenomenon of two components that bind to C1q directly or via one or more components, comprising the following steps:
(A) first introducing a C1q-containing solution;
(B) adding at least one donor component or a component group containing at least one donor component to the C1q-containing solution [the donor component can emit a signal after being excited by a light source. A component comprising at least one compound or group of compounds (donor group), which can bind to C1q directly or via one or more components from the group of components ],
(C) adding at least one acceptor component or a component group containing at least one acceptor component to the C1q-containing solution [the acceptor component is released by the compound or compound group described in (b) A component comprising at least one compound or group of compounds (acceptor group) capable of receiving and emitting a signal in the form of receiving and electromagnetic radiation, wherein the acceptor component is directly or one or more of the component groups Can bind to C1q via further components],
(D) exciting at least one compound or compound group (donor group) according to (b) with a light source;
(E) detecting electromagnetic radiation emitted by at least one compound or group of compounds (acceptor group) according to (c) to detect the binding phenomenon;
Including the above method.   The method of 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 (d) is a laser or a lamp.   The method according to claim 1 or 2, wherein the detection according to (e) is performed using a photomultiplier tube.   The method of claim 1, wherein the components of the component group described in (b) and / or (c) can be bound to CRP in a spatially continuous manner or to each other.   The method according to claim 2, wherein the components of the component group according to (b) and / or (c) can be bound spatially consecutively to C1q or to each other.   8. A method according to any one of claims 1 to 7, wherein the component is a polypeptide or comprises a polypeptide.   9. The method of claim 8, wherein at least one of the components is an antibody.   The method according to 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 CRP.   12. The method of claim 11, wherein the natural binding partner of CRP is C1q.   The method according to claim 12, wherein the component group according to claim 1 (a) or claim 1 (b) comprises C1q and an anti-C1q antibody.   The method of claim 2, wherein at least one of the components is a natural binding partner of C1q.   15. A method according to claim 14, wherein the natural binding partner is CRP.   The method according to claim 15, wherein the component group according to claim 2 (a) or claim 2 (b) comprises CRP and an anti-CRP antibody.   A signal is transmitted from at least one compound or compound group (donor group) described in (b) to at least one compound or compound group (acceptor group) described in (c) by emissionless energy transfer. The method according to claim 1 or 2.   The method of claim 17, wherein the emissionless energy transfer is fluorescence resonance energy transfer.   The method according to claim 1 or 2, wherein the signal is transmitted via singlet oxygen.   The at least one compound or compound group (donor group) according to claim 1 (b) or claim 2 (b) is a compound capable of converting triplet oxygen to singlet oxygen after excitation by a laser. Wherein at least one compound or group of compounds (acceptor group) according to claim 1 (b) or claim 2 (b) is at least one first compound excitable by singlet oxygen, and The at least one second compound capable of absorbing energy absorbed by the compound excitable by singlet oxygen in an emissionless manner and releasing it in the form of fluorescent radiation. Method.   21. The method according to claim 20, wherein the donor group and / or the acceptor group are localized on the particle.   The method of claim 21, wherein the average diameter of the particles is about 200 nm.   23. A method according to any one of claims 1 to 22 used for measuring the concentration of a CRP-containing solution having an unknown CRP content.   23. A method according to any one of the preceding claims, used to measure the concentration of a C1q-containing solution having an unknown C1q content.   25. The method according to claim 23 or 24, wherein the solution is serum, plasma, or serum or plasma diluted with physiological buffer or water.   26. The method according to claim 25, which is a diagnostic method for measuring the state of inflammation of an organism and / or the risk of cardiac arrest and / or stroke.   27. A method according to any one of the preceding claims, which is a HTS capable homogeneous method that can be performed as a one-step method.   Step (a) and / or step (b) and / or step (c) and / or step (d) to observe whether the test substance affects the binding phenomenon between CRP and the CRP binding component The method of claim 1, comprising the additional step of adding at least one test substance prior to the step.   Step (a) and / or step (b) and / or step (c) and / or step (d) in order to observe whether the test substance affects the binding phenomenon between C1q and the C1q binding component The method of claim 2, comprising the additional step of adding at least one test substance prior to.   30. The method according to claim 28 or 29, wherein the method is used to measure substances that affect the binding phenomenon in a modulating, inhibiting or activating manner. An HTS capable diagnostic analysis system for measuring a binding phenomenon between a CRP and a component that binds to CRP, comprising:
(A) At least one donor component or a component group including at least one donor component [the donor component is capable of emitting a signal after being excited by a light source ( A component comprising a donor group, which can bind to the CRP directly or via one or more components from the component group],
(B) at least one acceptor component or a group of components comprising at least one acceptor component [the acceptor component receives a signal emitted by the compound or group of compounds described in (A) and forms of electromagnetic radiation A component comprising at least one compound or group of compounds (acceptor group) that can be released in the CRP, wherein the acceptor component is directly or via one or more components of the group of components to the CRP Can be combined]
Including said diagnostic analysis system. An HTS capable diagnostic analysis system for measuring a binding phenomenon between C1q and a component binding to C1q, comprising the following components:
(A) At least one donor component or a component group including at least one donor component [the donor component is capable of emitting a signal after being excited by a light source ( A component comprising a donor group, which can be bound to C1q directly or via one or more components from the component group],
(B) at least one acceptor component or a group of components comprising at least one acceptor component [the acceptor component receives a signal emitted by the compound or group of compounds described in (A) and forms of electromagnetic radiation A component comprising at least one compound or group of compounds (acceptor group) that can be released in the form of an acceptor component to C1q directly or via one or more components of the group of components Can be combined]
Including said diagnostic analysis system. The following additional ingredients:
(C) Serum or plasma, or serum or plasma diluted with physiological buffer or water,
(D) a light source for exciting at least one compound or compound group (donor group) according to (A),
33. An HTS capable analysis system according to claim 31 or 32 comprising a detection system for detecting the emitted electromagnetic radiation of (E) (B). An HTS-capable analytical system for measuring active substances acting in a regulated manner on the interaction between components binding to CRP and CRP, comprising the following components:
(A ′) at least one donor component or a component group including at least one donor component [the donor component is capable of emitting a signal after being excited by a light source; (Donor group), which can bind to CRP directly or via one or more components from the group]
(B ′) at least one acceptor component, or a group of components comprising at least one acceptor component [the acceptor component receives and emits a signal emitted by the compound or group of compounds according to (A ′) and electromagnetic radiation A component comprising at least one compound or group of compounds (acceptor group) that can be released in the form of, wherein the acceptor component is directly or via one or more components of the group of components Can bind to CRP],
(C ′) The above analytical system comprising at least one test compound. An HTS-capable analytical system for measuring active substances acting in a regulated manner on the interaction between C1q and components binding to C1q, comprising the following components:
(A ′) at least one donor component or a component group including at least one donor component [the donor component is capable of emitting a signal after being excited by a light source; (The donor group), which can be bound to C1q directly or via one or more components from the group]
(B ′) at least one acceptor component, or a group of components comprising at least one acceptor component [the acceptor component receives and emits a signal emitted by the compound or group of compounds according to (A ′) and electromagnetic radiation A component comprising at least one compound or group of compounds (acceptor group) that can be released in the form of, wherein the acceptor component is directly or via one or more components of the group of components Can bind to C1q],
(C ′) The above analytical system comprising at least one test compound. 36. The HTS capable analysis system of claim 34 or 35, wherein the following additional components:
(D ′) a light source for exciting at least one compound or compound group (donor group) according to (A ′),
(E ′) An analytical system as described above, comprising a detection system for detecting the emitted electromagnetic radiation according to (B ′).

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