Device and method for purifying body fluids by removing angiotensin-specific antibodies

Abstract

The present invention relates to a device for purifying body fluids with the aim of removing certain antibodies e.g. from human or animal blood. The antibodies to be removed target angiotensins. The devices according to the invention are intended to remove these specific antibodies from the blood or the liquor and to thus alleviate certain illnesses, in particular long COVID and similar syndromes and their consequences. The invention also relates, inter alia, to methods for carrying out extracorporeal blood purification using the described devices.

Description

[0001] Device and method for purifying body fluids by separating angiotensin-specific antibodies

[0002] The present invention describes a device for purifying body fluids with the aim of removing certain antibodies from the human body. The antibodies to be removed are directed against angiotensins. Purification can be carried out, among other things, in the manner of a blood purification (apheresis).

[0003] There is evidence that some people develop antibodies directed against various angiotensins after certain viral infections (e.g., S ARS-CoV2). Such antibodies can cause long-lasting disorders and may be a cause of so-called "long COVID" diseases and similar pathological manifestations after viral infections, which are observed in a subset of infected individuals. The antibodies formed may not only be specifically directed against angiotensins but also against certain viral proteins, e.g., the spike protein. The device described in this document aims to remove these specific antibodies from the blood and thus alleviate the diseases described above and their consequences. Methods for performing extracorporeal blood dialysis using the described device are also described.

[0004] The state of the art includes the following publications:

[0005] WO 2020 / 127969 Al

[0006] WO 20197028006 Al

[0007] WO 2022 / 126119 A2

[0008] Briquez, Priscilla S. et al.: “Severe COVID-19 induces autoantibodies against angiotensin II that correlate with blood pressure dysregulation and disease severity,” Science Advances, Vol. 8, No. 40, pp. 1-10

[0009] The present invention is based on a technology aimed at removing antibodies from body fluids which are produced by the body as a result of an infection, e.g. with coronaviruses (in particular SARS-CoV1 & SARS-CoV2), but also other viruses, and which then develop a specificity against angiotensin II (angiotensinogen, angiotensin I [angiotensin 1-9 & angiotensin 1-7 are similar structures and would also be bound by the antibodies, among others]). This virus-induced specificity of the antibodies to the body's own angiotensins, such as angiotensin II, may result in a disruption of the renin-angiotensin-aldosterone system (RAAS) being induced by the binding of free angiotensins, such as angiotensin II. This disruption can be acutely life-threatening and / or long-lasting. This disorder could be a cause of “long Covid” and similar syndromes, such as post-vaccine syndrome or chronic SARS.The present invention aims to remove the formed anti-angiotensin antibodies from the body, e.g. by extracorporeal treatment of body fluids, in particular blood and cerebrospinal fluid. The body fluid is brought into contact with a solid phase-fixed angiotensin or one of the derivatives mentioned below. The antibodies bind to angiotensin II, angiotensin I or angiotensinogen or a derivative or a structure which contains the sequence of the various angiotensins, wherein the angiotensin or derivative thereof is coupled to a solid phase or matrix by means of a linker, optionally using a spacer. The linker / spacer system can be rigid or flexible. It is important that it enables the antibody to bind to the angiotensin and does not have a steric impairing effect. The binding can, for example, be to a hydrophilic or hydrated membrane.a solid phase or matrix, as is known from numerous other processes. For simplicity, we will refer to the term matrix from here on.

[0010] After blood collection (analogous to conventional dialysis), the angiotensin I / II / -ogen autoantibodies can be removed from the whole blood, blood plasma, or serum, and the serum, plasma, or blood can be reintroduced to the patient. The differential and specific removal of the angiotensin I / II / -ogen autoantibodies allows for gentle (no indifferent dialysis) and quantitatively controllable (variation of blood filtration efficiency based on various physiological factors) removal, allowing the RAAS to return to normal after a certain period of time. [The membrane can, for example, be easily implemented in commercially available plasma donation units.] The angiotensin I / II / -ogen is usually bound C-terminally, N-terminally, or in a mixed form (C- and N-terminally) to the membrane (whose surfaces are optimized), the linker / spacer, or the anchor (flexible or rigid if necessary).Alternatively, angiotensin can be bound via the functional groups of the angiotensin, for example, via the side groups of the amino acids. Covalent bonds are preferred, but ionic, dative, chelate, metal bonds, or any other type of immobilization are also possible. If necessary, degradation by amino and carboxy exoproteases can be prevented by C-terminal and / or N-terminal cappings to maintain long-term high efficiency of the membrane material (storage stability and durability under process stress).

[0011] The invention therefore relates firstly to a device for purifying body fluids by separating angiotensin-specific antibodies from the blood plasma and / or cerebrospinal fluid, containing at least one cartridge with matrix-bound angiotensins and a method for extracorporeal blood purification (apheresis) using this device.

[0012] For the purposes of this description, angiotensin-specific antibodies (also: anti-angiotensin antibodies) are understood to be antibodies that specifically bind to angiotensins, in particular to angiotensin I (angiotensin 1 to 10), angiotensin 1 to 9, angiotensin II (angiotensin 1 to 8), angiotensin 1 to 7, angiotensin III (angiotensin 2 to 8), and angiotensin IV (angiotensin 3 to 8), or to angiotensinogen. Angiotensin-specific antibodies also include antibodies that not only bind to angiotensin or angiotensinogen, but also exhibit cospecificity and are therefore also directed against other structures, in particular against viral proteins, e.g., the so-called spike protein of SARS-CoV2.

[0013] The device initially contains one or more angiotensins bound to a matrix. Angiotensins are well described in the literature. These are peptide hormones that, among other things, affect blood vessels and the heart rate, thus primarily influencing blood pressure. They can also influence the adrenal glands, particularly aldosterone levels. Other related effects of angiotensins have been described, such as kidney / adrenal function, hormone secretion, vasodilation and vasoconstriction, and influence on RAAS signaling cascades via the ATI receptor, AT2 receptor, and Mas receptor.It is therefore immediately clear that disturbances of the angiotensin system caused by the described antibodies can lead to a variety of symptoms, as have also been widely described in the literature for “long Covid” and similar syndromes, such as post-vaccine syndrome or chronic SARS.

[0014] Angiotensins that are particularly known are angiotensin I (angiotensin 1 to 10), angiotensin

[0015] 1 to 9, angiotensin II (angiotensin 1 to 8), angiotensin 1 to 7, angiotensin III (angiotensin

[0016] 2 to 8) and angiotensin IV (angiotensin 3 to 8). The device according to the invention can contain individual angiotensins or a mixture of different angiotensins. The sequences of the angiotensins mentioned are described in the specialist literature and therefore require no repetition here. Also according to the invention are angiotensin derivatives with parts of the above-mentioned angiotensin sequences, as well as derivatives in which the sequence sequence is inverted and / or in which amino acids have been exchanged, as well as glycosylated or otherwise chemically modified derivatives. Inverted sequence sequences are, for example, the sequence 3'-DRVYIHPFHL-5' (ie 5'-LHFPHIYVRD-3' in standard reading direction) instead of the usual sequence 5'-DRVYIHPFHL-3'.

[0017] Also included in the invention are topologically equivalent peptides or chemical structures. In each case, the ability to bind the anti-angiotensin antibodies defined above is crucial. Angiotensin I is particularly preferred for the compound according to the invention.

[0018] The angiotensins mentioned are bound to a matrix. The binding can be N-terminal and / or C-terminal, but can also be achieved through one of the other binding possibilities described above.

[0019] The solid phases known from the state of the art for comparable processes can be used as the solid phase for binding the angiotensins, in particular

[0020] - Gold nanoparticles

[0021] - Silicon dioxide

[0022] - glass beads

[0023] - Carbon beads

[0024] - metal beads

[0025] - Magnetic Beads

[0026] - polystyrene derivative

[0027] - Polycarbonate derivatives

[0028] - polyvinyl chloride)

[0029] - Polyacrylamide

[0030] - Polyethylene terephthalate (PET)

[0031] - Polyvinylidene fluoride (PVDF) derivatives

[0032] - Polyvinyl acetate (PVAc) derivatives

[0033] - Polyvinylpyrrolidone (PVP) derivatives

[0034] - Polyethylene oxide (PEO) derivatives

[0035] - Poly(ethyleneimine) derivatives (PEI)

[0036] - NHS-activated agarose / Sepharose / hydrogels - Agarose / Sepharose / hydrogels optionally with immobilized DADPA or NHS

[0037] - (CarboxyLink resin)

[0038] The matrix materials can be surface-modified to bind spacers or angiotensins (e.g., thiol-modified, amine-modified). Alginates and hydrogels are particularly preferred as matrices for the present invention.

[0039] The methods for binding angiotensins to a solid phase are sufficiently described in the state of the art, the necessary materials and reagents are largely commercially available, so that explanations are unnecessary at this point.

[0040] A linker must be present between the matrix and the angiotensin to bind the angiotensin or the spacer associated with the angiotensin to the matrix. For the purposes of the present invention, the term linker also includes groups sometimes referred to as "anchors," which serve to bind peptides and / or spacers to the matrix. Such linkers are known in the art. Examples include the following:

[0041] - Peptide derivatives (e.g. PEG peptide, PVA peptide)

[0042] - Compounds containing disulfide bonds (e.g. BMPER)

[0043] - Derivatives with amine-reactive groups (e.g. DCC, DBCO)

[0044] - Derivatives with carbonyl-reactive groups (e.g. DCC, DBCO)

[0045] - Derivatives with thiol-reactive groups (e.g. DTSSP, DTSSL)

[0046] - DADPA (diaminodipropylamine)

[0047] - Hydrazide-containing compounds

[0048] - Derivatives of hydroxysuccinimide esters (e.g. DBCO, BMPER)

[0049] - Derivatives with reactive carboxyl groups (e.g. EDC, NHS)

[0050] - Derivatives with epoxy-reactive groups (e.g. EDC, NHS)

[0051] - Click chemistry derivatives (e.g. alkyne azide)

[0052] - Derivatives with photolabile groups (e.g. DMTP)

[0053] - Amino acids

[0054] - Peptides (e.g. Gly-Gly-Cys)

[0055] - Polypeptides

[0056] - Proteins

[0057] - Disulfide bonds

[0058] - Amine-reactive groups (e.g. NHS esters) - Carbonyl-reactive groups (e.g. NHS amides)

[0059] - Thiol-reactive groups (e.g. May eimide)

[0060] - Hydrazide

[0061] - Hydroxysuccinimide ester (NHS ester)

[0062] - Carboxyl-reactive groups (e.g. EDC)

[0063] - Epoxy-reactive groups (e.g. EDC)

[0064] - Photolabile groups / proteins (e.g. DMT, MeNPOC, NPPOC) see also Further reading, below)

[0065] - Click chemistry (e.g. azide-alkyne cycloaddition)

[0066] Explanation of click chemistry: In click chemistry, chemical reactions such as copper-catalyzed azide-alkyne cycloaddition (CuAAC) or strain-assisted azide-alkyne cycloaddition (SPAAC) are used to create covalent bonds between the protein and the surface.

[0067] Particularly preferred linkers are lectins as well as biotin / avidin or streptavidin and DADPA linkers or NHS linkers.

[0068] To avoid contamination of body fluids during extracorporeal washing by angiotensins, the bond between matrix and angiotensin or matrix and spacer as well as between spacer and angiotensin is preferably designed as a covalent bond.

[0069] The methods for coupling angiotensins to a matrix by means of a linker are sufficiently described in the prior art and do not require any explanation here.

[0070] A spacer can also be present between the matrix and the angiotensin. A spacer is generally a molecule or group of molecules that spatially separates the protein from the matrix and creates a gap between the protein and the matrix surface. Rigid or flexible peptides or proteins are particularly suitable for this purpose. Numerous other spacers are known from the state of the art and can be readily used.

[0071] Examples of spacers include:

[0072] - Polyethylene glycol (PEG) derivatives (e.g. PEG-thiol, PEG-amine)

[0073] - Polyvinyl alcohol (PVA) derivatives (e.g. PVA thiol, PVA amine) - Polysaccharide derivatives (e.g. dextran sulfate, chitosan)

[0074] - Polyamidoamine (PAMAM) dendrimers - Polyamidoxime dendrimers (PAOx)

[0075] - Polyethyleneimine (PEI) derivatives

[0076] - Polyethylene oxide (PEO) derivatives

[0077] - Polyvinylpyrrolidone (PVP) derivatives

[0078] - Polyvinyl acetate (PVAc) derivatives

[0079] - Polyvinyl chloride (PVC) derivatives

[0080] - Polyvinylidene fluoride (PVDF) derivatives

[0081] - Polyacrylamide (PAAm) derivatives

[0082] - Derivatives of polymethacrylate (PMMA)

[0083] - Polyvinylpyrrolidone (PVP)

[0084] - Amino acids

[0085] - Peptides (e.g. Gly-Gly-Cys)

[0086] - Polypeptides

[0087] - Proteins

[0088] - Polyethylene oxide (PEO)

[0089] - Poly(ethyleneimine) (PEI)

[0090] - Polyvinyl alcohol (PVA)

[0091] - Dextran

[0092] The entire process for producing the device according to the invention can in principle be carried out as described for other peptides as antibody capture devices in similar applications, e.g. in DE 1953864 A1.

[0093] It is known that peptides, polypeptides, proteins, or fusion proteins can be degraded by proteases present in blood plasma. To prevent such degradation, the construct consisting of spacers, linkers, peptides, polypeptides, proteins, or fusion proteins can be protected from degradation by proteases by N-terminal and / or C-terminal cappings.

[0094] Known cappings are listed below:

[0095] A) N-terminal cappings:

[0096] 1 Acetylation - Addition of an acetyl group to the amino group of the N-terminus of a protein, which can affect the stability, localization, and function of the protein. 2 Formylation - Addition of a formyl group to the N-terminus of a protein, which occurs in bacterial proteins and can impair immune recognition of the protein.

[0097] 3 Myristoylation - Addition of a myristic acid molecule to the N-terminus of a protein, which can affect membrane association and signal transduction.

[0098] 4 Palmitoylation - Addition of a palmitic acid molecule to the N-terminus of a protein, which can affect membrane association and stability.

[0099] 5 Glycosylation - Addition of a sugar molecule to the N-terminus of a protein, which can affect folding, stability and function.

[0100] 6 Phosphorylation - Addition of a phosphate group to the N-terminus of a protein, which can affect signaling, activity and localization.

[0101] 7 Methylation - Addition of a methyl group to the N-terminus of a protein, which can affect protein-protein interactions and transcriptional regulation.

[0102] 8 Sumoylation - Addition of a small ubiquitin-like modifier (SUMO) to the N-terminus of a protein, which can affect protein-protein interactions and localization.

[0103] 9 Ubiquitination - Attachment of ubiquitin to the N-terminus of a protein, which can affect protein degradation, localization, and signal transduction.

[0104] 10 Acylation - Addition of an acyl group (such as acetyl, palmitoyl, or myristoyl) to the N-terminus of a protein, which can affect membrane association and stability.

[0105] 11 N-terminal truncation - Removal of the N-terminal amino acids from a protein, which can affect the folding, stability and activity of the protein.

[0106] 12 Proteolytic cleavage - enzymatic removal of a peptide bond at the N-terminus of a protein, which can activate or inactivate the protein.

[0107] 13 Oxidation - Addition of an oxygen molecule to the N-terminus of a protein, which can affect stability and activity.

[0108] 14 Sulfation - Addition of a sulfate group to the N-terminus of a protein, which can impair protein-protein interactions and function.

[0109] 15 Nitrosylation - Addition of a nitric oxide molecule to the N-terminus of a protein, which can affect protein activity and signal transduction.

[0110] 16 Glutathionylation - Addition of a glutathione molecule to the N-terminus of a protein, which can impair protein-protein interactions and function. 17 Proline isomerization - Isomerization of the N-terminal proline residue of a protein, which can impair protein folding and stability.

[0111] 18 Formylation-dependent initiation - Initiation of translation with a formylated methionine residue at the N-terminus of a protein, which occurs in bacterial proteins.

[0112] 19 Pyroglutamate formation - Formation of a pyroglutamate residue at the N-terminus of a protein, which can affect protein stability and function.

[0113] 20 The N-terminal attachment of amino acids to peptides, polypeptides or proteins, whereby the degradation of the extended construct containing spacers, linkers, peptides, proteins or fusion proteins and / or polypeptides is delayed and thus also the degradation of the angiotensins.

[0114] 21 N-terminal binding to the matrix (cross-linking or loop formation)

[0115] B) C-terminal cappings:

[0116] 1 Amidation - Conversion of the C-terminal carboxyl group of a protein into an amide group, which can affect the stability, activity and localization of the protein.

[0117] 2 Glycosylation - Addition of a sugar molecule to the C-terminus of a protein, which can affect folding, stability and function.

[0118] 3 Phosphorylation - Addition of a phosphate group to the C-terminus of a protein, which can affect signaling, activity and localization.

[0119] 4 Methylation - Addition of a methyl group to the C-terminus of a protein, which can affect protein-protein interactions and transcriptional regulation.

[0120] 5 Sumoylation - Addition of a small ubiquitin-like modifier (SUMO) to the C-terminus of a protein, which can influence protein-protein interactions and localization.

[0121] 6 Ubiquitination - Attaching ubiquitin to the C-terminus of a protein, which can affect protein degradation, localization, and signal transduction.

[0122] 7 C-terminal truncation - Removal of the C-terminal amino acids from a protein, which can affect the folding, stability and activity of the protein.

[0123] 8 Proteolytic cleavage - enzymatic removal of a peptide bond at the C-terminus of a protein, which can activate or inactivate the protein. 9 Sulfation - addition of a sulfate group to the C-terminus of a protein, which can affect protein-protein interactions and function.

[0124] 10 Nitrosylation - Addition of a nitric oxide molecule to the C-terminus of a protein, which can affect protein activity and signal transduction.

[0125] 11 Glutathionylation - Attachment of a glutathione molecule to the C-terminus of a protein, which can impair protein-protein interactions and function.

[0126] 12 Ribosomal skipping - skipping of C-terminal amino acids during translation, which can affect the activity and stability of proteins.

[0127] 13 Proline isomerization - Isomerization of the C-terminal proline residue of a protein, which can affect protein folding and stability.

[0128] 14 Peptidylarginine deiminase (PAD) modification - Conversion of arginine residues to citrulline residues at the C-terminus of a protein by PAD enzymes, which can impair protein function and immune recognition.

[0129] 15 Glutamic acid decarboxylation - Decarboxylation of the C-terminal glutamic acid residue of a protein, which can impair the activity and function of the protein.

[0130] 16 Asparagine deamidation - Deamidation of the C-terminal asparagine residue of a protein, which can impair the function and stability of the protein.

[0131] 17 Lysine carboxylation - Addition of a carboxylic acid group to the C-terminal lysine residue of a protein, which can impair protein-protein interactions and function.

[0132] 18 Tyrosine sulfation - Addition of a sulfate group to the C-terminal tyrosine residue of a protein, which can impair protein-protein interactions and function.

[0133] 19 C-terminal thioesterification - Addition of a thiol group to the C-terminal carboxyl group of a protein, which can influence protein-protein interactions and function. 0 The C-terminal attachment of amino acids to peptides, polypeptides, or proteins, thereby delaying the degradation of the extended construct containing spacers, linkers, peptides, proteins, fusion proteins, and / or polypeptides, and thus also the degradation of angiotensins. 1 C-terminal binding to the matrix (crosslinking or loop formation). The methods for capping peptides, polypeptides, proteins, and fusion proteins are sufficiently described in the prior art and require no further explanation here.

[0134] For the application according to the invention, the matrix-coupled angiotensins are arranged, for example, in a cartridge. Such cartridges are already known from the prior art for other applications, in particular blood dialysis (apheresis), and are commercially available. The cartridges are usually made of plastic, glass, or a neutral metal, are cylindrical in shape, and have a size of 20 to 150 ml. The cartridges have two connections on the front sides and can be integrated into conventional blood dialysis devices. The cartridges can have diffusers, particularly at the inlet, which evenly distribute the flow in the cartridge and ensure turbulent mixing. Of course, several cartridges can be used consecutively if necessary. Likewise, several cartridges can be operated in parallel to enable a higher throughput of the system.

[0135] A first embodiment of the invention is a blood dialysis procedure: To perform extracorporeal blood dialysis, blood is first drawn from the patient. The blood plasma is extracted from the drawn whole blood and passed through the cartridge containing the matrix-bound angiotensins. As it passes through the cartridge, the anti-angiotensin antibodies bind to the angiotensins. The blood plasma emerging from the cartridge is therefore essentially free of these anti-angiotensin antibodies. The plasma is combined with the other blood components separated in the first step (particularly erythrocytes) and returned to the patient. This procedure can be designed as a continuous process. The patient is typically subjected to the described extracorporeal blood dialysis for two to five hours.

[0136] In some embodiments of the invention, the removal of anti-angiotensin antibodies from the blood serum is carried out.

[0137] In specially adapted devices, which are known in principle, the treatment of whole blood, ie without prior extraction of the blood plasma, is also possible.

[0138] After the described extracorporeal hemodialysis, the patient is initially free of the anti-angiotensin antibodies described above. The side effects triggered by the antibodies typically subside, particularly the renin-angiotensin-aldosterone system (RAAS) described in the introduction, which returns to normal. The invention is therefore suitable for the treatment of patients with "long COVID syndrome," but also for the treatment of similar conditions, such as post-vaccine syndrome (after COVID vaccination) or chronic SARS-CoV-2, which have comparable causes. If necessary, the procedure can be repeated multiple times.

[0139] The use of the invention can be advantageous in the treatment of acute COVID patients. With timely removal of anti-angiotensin antibodies, the aforementioned disorders can be prevented at an early stage. The use of the invention according to the invention can also be used in acute, life-threatening disorders of the renin-angiotensin-aldosterone system (RAAS), thereby eliminating or minimizing the RAAS system disorders. The invention can also be used preventively in acute infections to prevent any disruption of the RAAS system at an early stage.

[0140] The treatments according to the invention, such as hemodialysis, can be performed in combination with other clinical treatments, such as extracorporeal ventilation (ECMO) or dialysis. The device according to the invention can, for example, be arranged upstream or downstream of the oxygenator or dialyzer in the extracorporeal circuit, or alternatively, be directly connected to the oxygenator or dialyzer at the blood inflow or outflow, wherein the device is configured to be perfused with whole blood.

[0141] A particularly advantageous feature is that the removal of angiotensin-specific antibodies is gentle, without burdening the patient with additional medication. The removal of antibodies according to the invention can also be performed slowly, allowing a gradual return of the affected physiological control circuits to normal, which places less strain on the affected body than a sudden change.

[0142] A second embodiment of the invention is cerebrospinal fluid (CSF) washing. To perform extracorporeal CSF washing, cerebrospinal fluid (CSF) is first withdrawn from the affected patient, e.g., from the subarachnoid space or spinal canal, by means of a lumbar puncture. The CSF is passed through the cartridge containing the matrix-bound angiotensins. As it passes through the cartridge, the specific antibodies bind to the angiotensins. The CSF exiting the cartridge is therefore essentially free of these anti-angiotensin antibodies and is returned to the patient. This procedure can be designed as a continuous process. The patient is typically subjected to the described extracorporeal CSF washing for two to five hours. If necessary, the procedure can be repeated several times.

[0143] The procedures described are similar to other immunoadsorption procedures, such as those used to remove autoimmune antibodies from the body.

[0144] The cartridges loaded with angiotensin-specific antibodies after the procedure can optionally be subjected to regeneration. For this purpose, either the bound antibodies are separated from the angiotensins using standard techniques, or the linker bond is broken, separating the angiotensins. Chaotropic salts and / or detergents can be used for this purpose. When using a biotin / avidin or streptavidin linker, it is also possible to rinse with a biotin solution, whereby the solution has a concentration 10–100 times higher than the bound biotin. In the case of bonds between carbohydrates and lectin, carbohydrate solutions can cleave the bond through competitive reactions. Alternatively or additionally, pH changes or chaotropic salts can lead to detachment. When using light-dissociable linkers, release can occur through light irradiation.The individual procedures are standard practice and will therefore not be explained further here.

[0145] After a final rinse, the cartridge can be reinserted.

[0146] By separating the anti-angiotensin antibodies from cartridges loaded with them, the antibodies can also be obtained commercially, for example for research purposes.

[0147] In a further alternative embodiment of the invention, the matrix-coupled angiotensins are arranged in a vascular implant. Such vascular implants are known in principle and are usually made of PTFE (Teflon), polyethylene terephthalate (Dacron), or polyurethane, with the angiotensin coupled to the inner wall as a solid phase. Vascular implantation can provide prolonged blood dialysis.

[0148] In a further development of this invention, the vascular implant can also be regenerated intravascularly. External ports are provided for this purpose. For the purpose of regeneration, the blood flow through the vessel must be clamped off. The ports allow the rinsing fluids or regeneration fluids to be introduced into the implant, analogous to the regeneration of the aforementioned cartridges. After regeneration is complete, the implant must be sufficiently rinsed, for example, with physiological saline solution. After the ports are closed, blood flow can be restored.

[0149] The treatment options described above can also be applied to mammals affected by long COVID and similar syndromes. Similarly, the methods for washing body fluids can also be used to separate anti-angiotensin antibodies from cell cultures.

[0150] Description of the characters

[0151] Figure 1 schematically shows the first application of the device according to the invention: First, blood is drawn from the patient. The blood plasma is extracted from the drawn whole blood and passed through the cartridge containing the solid-phase-bound angiotensins. The plasma is combined with the other blood components (e.g., erythrocytes) separated in the first step and returned to the patient. This procedure can be designed as a continuous process.

[0152] Figure 2 schematically shows matrix-bound angiotensin, which is bound to the matrix (e.g., hydrogel matrix) via a spacer, which in turn is bound to the matrix via a linker. The linker is not shown in the graphic. The bound antibody is also shown schematically.

[0153] Figure 3 shows a schematic of several angiotensin molecules bound to a matrix via a spacer, which in turn is bound to a matrix via a linker. The linker is not shown in the graphic. The number of spacers and angiotensins can be higher than that shown schematically here, and a spacer can also be attached after an angiotensin in a different way than shown in the graphic. There is another spacer between the two angiotensin molecules shown here as an example. The advantage of several angiotensin molecules bound to one point in the matrix is, on the one hand, that a higher density of angiotensin molecules can be achieved, and on the other hand, the degradation of the angiotensin is delayed, since the degradation of the angiotensin contained in the construct by exopeptidases only occurs later. This can be achieved by attaching amino acids, peptides, polypeptides, or fusion proteins to the angiotensins.

[0154] Figure 4 shows a schematic of a matrix-bound angiotensin which is protected from degradation by exopeptidases by a capping. Capping can also be the attachment of amino acids, peptides, polypeptides or fusion proteins to the angiotensins. Figure 5 shows a schematic of several angiotensin molecules which form a "loop" with the spacers, i.e. are bound to the matrix in a loop-like manner. In this embodiment of the invention, the angiotensins are better protected from degradation by exopeptidases. The matrix acts as a capping in a sense, since the N-terminus and C-terminus are bound to the matrix and neither the N-terminus nor the C-terminus is exposed, which can be considered necessary for degradation by exopeptidases.

[0155] Figure 6 schematically shows an embodiment of the invention in which the construct consisting of a spacer and angiotensin is arranged in a cross-linking manner between two matrix surfaces. Analogous to the embodiment in Figure 5, no capping is required here to protect against exopeptidases, since the second matrix surface acts as a capping agent. The matrix acts as a capping agent in a sense, since the N-terminus and C-terminus are bound to the matrix, leaving neither the N-terminus nor the C-terminus exposed, which can be considered necessary for degradation by exopeptidases.

[0156] Further reading

[0157] General publications on protein immobilization https: / / www.sciencedirect.com / topics / biochemistry-genetics-and-molecular-biology / protein- immobilization.

[0158] Publications for linkers https: / / broadpharm.com / product-categories / adc-linkers https: / / www.pepscan.com / custom-peptide-svnthesis / peptide-modifications / linkers-spacers /

[0159] Li G, Huang Z, Zhang C, Dong BJ, Guo RH, Yue HW, Yan LT, Xing XH. Construction of a linker library with widely controllable flexibility for fusion protein design. Appl Microbiol Biotechnol. 2016 Jan;100(l):215-25. doi: 10.1007 / s00253-015-6985-3. Epub 2015 Sep 22. PMID: 26394862. (https: / / pubmed.ncbi.nlm.nih.gov / 26394862 / )

[0160] Ziliang Huang, Chong Zhang, Xin-Hui Xing, Chapter Two - Design and construction of chimeric linker library with controllable flexibilities for precision protein engineering, Editor(s): Maarten Merkx, Methods in Enzymology, Academic Press, Volume 647, 2021, Pages 23-49, doi: 10.1016 / bs.mie.2020.12.004. ISSN 0076-6879, ISBN 9780128208182 (https: / / www.sciencedirect.coni / science / article / abs / pii / S0076687920303700) https: / / www.kbdna.com / publishinglab / lnkr Federica Rusmini, Zhiyuan Zhong, and Jan Feijen. Protein Immobilization Strategies for

[0161] Protein Biochips. Biomacromolecules. 2007 8 (6), 1775-1789

[0162] DOI: 10.1021 / bm061197b (https: / / pubs.acs.org / doi / pdf / K' 102 l / bmlv I l °7h)

[0163] Ziliang Huang, Gang Li, Chong Zhang, Xin-Hui Xing, A study on the effects of linker flexibility on acid phosphatase PhoC-GFP fusion protein using a novel linker library, Enzyme and Microbial Technology, Volume 83, 2016, Pages 1-6, ISSN 0141-0229, doi: / 10

[0164] (https: / / Dihub)

[0165] Datenbanken für Linker https : / / www.ibi .vu.nl / program s / linkerdbwww / http: / / parts.igem.org / Protein domains / Linker

[0166] Lectin Bindung: itml

[0167] Zhou SM, Cheng L, Guo SJ, Zhu H, Tao SC. Lectin microarrays: a powerful tool for glycan- based biomarker discovery. Comb Chem High Throughput Screen. 2011 Sep;14(8):711-9. doi: 10.2174 / 138620711796504398. PMID: 21933110.

[0168] (https: / / www.researchgate.net / publication / 51657008 Lectin Mi croarrays A Powerful Tool mfor Glycan-Based Biomarker Discovery / figures?lo=l)

[0169] Photolabile linkers

[0170] Light-dissociable syntactic organelles / light-dissociable optogenetic proteins, see, for example: Schematic of optoCluster and PixELL formation; Zhao et al. (2019)

[0171] Proceedings

[0172] Immobilization using NHS-activated agarose https: / / de.vwr.com / store / product / 18502975 / nhs-activated-agarose-piercetm

[0173] Immobilization using diaminodipropylamine (DADPA)-activated agarose https: / / www.thermofisher.com / order / catalog / product / 20266

[0174] Autoimmune adsorption process https: / / www.freseniusmedicalcare.com / fileadmin / data / de / pdf / Healthcare Professional s / IA / IA

Claims

Claims 1. Device for washing body fluids by separating angiotensin-specific antibodies from the body fluid, containing at least one cartridge with matrix-bound angiotensins.

2. Device for washing body fluids according to claim 1, wherein the angiotensins comprise angiotensin II, angiotensinogen, angiotensin I, angiotensin 1-9 and / or angiotensin 1-7 and their derivatives.

3. Device for washing body fluids according to claim 1, wherein the angiotensins are bound to a matrix by linkers.

4. A device for washing body fluids according to claim 1, wherein a spacer is arranged between the angiotensins and the matrix.

5. Device for washing body fluids according to claim 1, wherein the angiotensins contained in the construct of linker, spacer, peptides, polypeptides and / or fusion proteins are protected from degradation by proteases by N-terminal and / or C-terminal cappings.

6. Device for washing body fluids according to claim 1, wherein the angiotensins are bound N-terminally and / or C-terminally 7. Device for washing body fluids according to claim 1, containing a hydrogel matrix, a peptide spacer, wherein the peptide spacer is bound by NHS or DADPA and angiotensin I coupled to the peptide spacer.

8. Device for washing body fluids according to claims 1 to 7, wherein the device consists of a cartridge containing a solid phase to which angiotensins are bound by linkers and which has two ports as inlet and outlet of body fluids.

9. Device for washing body fluids according to claims 1 to 7, wherein the device consists of a vascular implant which serves as a solid phase to which angiotensins are bound by linkers.

10. Device for washing body fluids according to claim 9, wherein the vascular implant contains two ports by means of which a passage of rinsing fluids for regeneration is possible. REVISED SHEET (RULE 91) ISA / EP 11. A method for the extracorporeal washing of body fluids by separating angiotensin-specific antibodies from the blood, comprising the following steps: a) taking whole blood from a patient b) optionally obtaining plasma or serum from the whole blood c) passing the blood serum, blood plasma or whole blood through at least one cartridge containing matrix-bound angiotensins of claims 1-8 d) optionally reuniting the purified blood plasma or blood serum with the blood components separated in step b e) returning the purified whole blood to the patient.

12. A method for extracorporeal washing of body fluids by separating angiotensin-specific antibodies from the blood according to claim 11, wherein the method is designed as a continuous method.

13. A method for the extracorporeal washing of body fluids by separating angiotensin-specific antibodies from the cerebrospinal fluid, comprising the following steps: a) taking cerebrospinal fluid from the subarachnoid space or the spinal canal of a patient b) passing the cerebrospinal fluid through at least one cartridge containing matrix-bound angiotensins of claims 1-8 c) returning the purified cerebrospinal fluid to the patient.

14. A method for extracorporeal washing of body fluids by separating angiotensin-specific antibodies from the cerebrospinal fluid according to claim 13, wherein the method is designed as a continuous method.

15. A method for extracorporeal washing of body fluids according to claim 11-14, wherein the cartridge is regenerated by separating the anti-angiotensin antibodies.

16. A method for obtaining anti-angiotensin antibodies by separating the anti-angiotensin antibodies from the cartridges according to claims 1-8 during or after the extracorporeal washing of body fluids according to claims 11-14.

17. Method for washing cell cultures by separating angiotensin-specific antibodies from the cell culture supernatant with the following steps REVISED SHEET (RULE 91) ISA / EP a) Taking cell culture supernatant from a cell culture containing anti-angiotensin antibodies b) Passing the cell culture supernatant through at least one cartridge containing matrix-bound angiotensins c) Returning the purified cell culture supernatant to the cell culture.

18. A method for treating or preventing long COVID, post-vaccine syndrome, chronic SARS, or disorders of the renin-angiotensin-aldosterone system (RAAS), comprising separating angiotensin-specific antibodies using a device according to claims 1-10.

19. A method for treating or preventing long COVID, post-vaccine syndrome, chronic SARS, or disorders of the renin-angiotensin-aldosterone system (RAAS), comprising performing a method according to claims 11-14. REVISED SHEET (RULE 91) ISA / EP

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