Use of alkali metal hydroxide solution for single sampling column regeneration

Abstract

The present invention relates to the use of alkali metal hydroxide for regenerating an apheresis column for removing CRP by affinity chromatography, as well as a simplified regeneration method for an apheresis column for removing CRP by affinity chromatography under the use of an alkali metal hydroxide solution and configuring an apheresis device to tolerate an alkali metal hydroxide solution and allow regeneration of an apheresis column for removing CRP by affinity chromatography in a continuous operation.

Description

Technical Field

[0001] The present invention relates to the use of an alkali metal hydroxide solution for regenerating a single sampling column, the single sampling column is particularly a single sampling column used for removing CRP by affinity chromatography, and relates to a simplified regeneration method of the single sampling column used for removing CRP by affinity chromatography under the use of an alkali metal hydroxide solution, the alkali metal hydroxide solution is preferably a sodium hydroxide solution, and the single sampling device is configured to tolerate the alkali metal hydroxide solution. Background Art

[0002] According to the World Health Organization (WHO), approximately 17 million people died from cardiovascular disease in 2008. This makes cardiovascular disease the most common cause of death among non-communicable diseases, accounting for approximately one-third of all deaths worldwide each year. It is estimated that by 2030, this number will increase to approximately 23 million deaths per year.

[0003] Cardiovascular disease is and continues to be the leading cause of death worldwide and also represents a huge medical expense for national health systems and health insurance companies. Two of the most common and devastating manifestations of cardiovascular disease are atherosclerosis and thrombosis, which in turn are particularly causative factors for myocardial infarction and stroke.

[0004] In recent years, great progress has been made in the treatment of cardiovascular diseases. This progress has been made possible not only by the increasing understanding of pathogenic mechanisms, but also by the early identification of patients at risk. In fact, the identification of disease risk and early treatment are important features of modern medical practice. Over the past 25 years, various factors and clinical parameters have been identified that are associated with the current disease state or the likelihood of future cardiovascular disease. These risk factors may be measurable biochemical or physiological parameters, such as serum cholesterol, HDL, LDL, and fibrinogen levels, but may also include behavioral patterns, such as obesity and smoking. In cases where a risk factor is not only indicative of the disease or its development, but is actually causally related to the development of the disease, therapeutic manipulation of that risk factor can affect the course of the disease or reduce the risk of its development.

[0005] As an acute phase protein, CRP is part of the innate immune system and is formed in the liver and released into the blood during inflammatory responses. The formation of CRP is mainly induced by cytokines expressed during acute or chronic inflammatory responses. The strongest stimulus for CRP formation is interleukin 6 (IL-6). Therefore, the levels of CRP and IL-6 in the blood are indicators of local or systemic inflammatory responses. Chronic inflammation is speculated to be one of the fundamental and supporting pathological phenomena of cardiovascular disease. In this context, it is increasingly believed that CRP can not only predict cardiovascular disease, but also has a causal relationship with the occurrence of cardiovascular disease or may affect its course.

[0006] The normal value of CRP in human blood varies from person to person, but on average is about 0.8 mg CRP per liter of blood, but in the case of acute or chronic inflammatory response, it can rise to more than 100 mg CRP per liter of blood (for example, bacterial infection, atherosclerosis, myocardial infarction). Since the half-life of CRP in the blood (about 19 hours) is constant, it is independent of the patient's health status, and the synthesis rate of CRP is solely responsible for regulating the CRP level in the blood (Pepys & Hirschfield, J. Clin. Invest., 2003, 111: 1805-1812). Therefore, the substantial increase in CRP synthesis under acute pathological conditions places special requirements on the treatment method of removing CRP from patients (high-risk or acute patients), because a large amount of CRP must be removed to reduce the blood CRP level to a normal value. Therefore, a particularly effective device and boundary conditions for removing CRP from the patient's blood are required.

[0007] Extracorporeal apheresis is a process for physically removing substances from blood or plasma by filtration, precipitation or adsorption. The term therapeutic apheresis generally refers to medical procedures for removing pathogenic components from circulating blood. Removal of pathogenic substances from blood or plasma by extracorporeal apheresis using extracorporeal circulation has become a routine clinical practice treatment for many diseases. For example, immunoadsorption can be used to specifically remove antibodies and circulating immune complexes from patient plasma.

[0008] For the therapeutic in vitro removal of C-reactive protein (CRP) from blood or plasma, CRP apheresis columns are known from the prior art which allow the selective removal of CRP from blood or plasma, wherein these columns contain a matrix substrate material (e.g. as agarose) with phosphorylcholine or a phosphorylcholine derivative as ligand to which CRP is covalently coupled.

[0009] European patent application EP 3 020 726 A1 discloses a column material functionalized with ω-phosphonooxy-alkylammonium groups and / or with ω-ammonium alkoxy-hydroxyphosphoryloxy groups for the Ca2+ transport of CRP. 2+ The advantage of using such phosphorylcholine derivatives for CRP apheresis is that CRP can be selectively removed from other plasma components (e.g. other plasma proteins) by highly specific ligands. One reason for this is that CRP is bound to the Ca2+ of phosphorylcholine or its derivatives. 2+ A dependent binding mechanism is used to adsorb CRP. The only other substances that also bind to phosphorylcholine are antibodies against phosphatidylcholine. The loss of other plasma proteins such as gamma-globulin is therefore minimized.

[0010] For therapeutic use of extracorporeal apheresis by an extracorporeal circulation system, reusable adsorbers are usually used, since apheresis is performed in multiple cycles during a course of treatment. Reusable adsorbers consisting of a housing filled with a carrier substance and binding factors coupled to the carrier substance are known from the prior art. Reusable adsorbers are usually regenerable, because after a certain amount of plasma, which depends on the concentration of the substance to be adsorbed, the adsorber is "saturated" and the binding of the substance no longer occurs. Two adsorbers are usually used for one treatment. While the plasma is pumped through one adsorber, the second adsorber is regenerated at the same time. In this process, the adsorber is rinsed of the binding substances with various regeneration solutions, thereby preparing it again for a new plasma load. The number of regenerations allowed is pre-specified by the manufacturer. Reusable adsorbers can only be used for the same patient. To prevent bacterial growth in the adsorber, it must be filled with a storage solution at the end of each treatment and must be rinsed before each new treatment. However, a lot of costs can be saved by reusing them. The operation of existing devices (combinations of two or more medical devices) is very complicated and demanding. In addition, these devices are rarely used overall.

[0011] An apheresis column is used for one patient in multiple treatment courses. For this purpose, the apheresis column is stored in a storage solution after treatment. In the prior art, at the end of the apheresis treatment, the apheresis column is first treated with a regeneration solution, then rinsed with a rinse solution such as physiological saline, and finally a storage solution such as a 0.04% polyhexamethylene biguanide solution (PHMB-solution) or a solution of sodium azide and PBS (phosphate buffered saline) is introduced into the apheresis column. Therefore, the storage step involves a lot of time investment by clinical staff. If the regeneration solution and the storage agent are the same solution, two additional steps for storing the apheresis column can be avoided.

[0012] Typically, in therapeutic apheresis, the patient's blood is first mixed with an anticoagulant (such as citrate or heparin) and separated into plasma and cellular components using a blood centrifuge or plasma filtration device. The plasma is then passed through an apheresis column and pathogenic substances are removed by adsorption. The processed plasma is then recombined with the cellular components and returned to the patient. During the regeneration process, the apheresis column is separated from the extracorporeal circulation system.

[0013] In apheresis for selectively removing CRP from plasma, an average of 6000 mL of plasma is processed in 6 to 12 cycles in one course of treatment. Including the regeneration of the apheresis column, processing 6000 mL of plasma typically requires about 4-5 hours. Therefore, the regeneration of the apheresis column is a time factor that affects the total duration of a course of treatment. Therefore, it is desirable to keep the regeneration time to a minimum, particularly since the apheresis column cannot be used to remove CRP from blood or plasma during the regeneration process.

[0014] For the regeneration of apheresis columns, especially CRP apheresis columns, glycine / HCl or EDTA solutions are used in the prior art, which denature bound proteins by a pH jump to the acidic range or release the bond between the adsorbed molecules to be removed (especially CRP) and the support material of the apheresis column by complexing the bound cations (especially calcium). In the prior art, no compounds are known that can separate the molecules to be removed (especially CRP) bound to the support material of the apheresis column by other mechanisms.

[0015] DE 4338858 C1 discloses a device for regenerating a single sampling column. DE 4338858 C1 teaches the use of a reservoir that temporarily stores plasma during the regeneration of the single sampling column. The regeneration of the single sampling column is performed by a combination of glycine / HCl, PBS and NaCl solutions known in the prior art. In addition, DE 4338858 C1 does not disclose a bypass line that allows the plasma flow to bypass the single sampling column during its regeneration and be diverted.

[0016] European patent application EP 3 020 726 A1 discloses the use of citrate solutions in the purification of CRP using affinity chromatography with phosphorylcholine and its derivatives. Binding buffer pH 8.0 (0.1 M Tris, 0.2 M NaCl, 2 mM CaCl2) and elution buffer pH 8.0 (EDTA) or regeneration solution pH 2.8 (glycine- / HCl) known in the prior art are used for the chromatography.

[0017] The inventors of the present invention have found that during regeneration with glycine / HCl, in particular during regeneration of a single sampling column used for affinity chromatography to remove CRP, a protein layer is formed around the matrix particles (e.g., agarose particles). This may be due to precipitation of acidic proteins. If the patient's blood to be purified contains a high concentration of DNA / RNA free from the cells, this effect may be enhanced. The formation of a protein layer in the single sampling column obscures the binding sites and reduces the performance of the single sampling material. The original state cannot be restored by known measures, such as further regeneration attempts with glycine / HCl solutions. So far, this problem has not been described in the prior art. As the damage to the single sampling column progresses, the patient's treatment time increases, and the patient's pain time also increases. In addition, damaged single sampling columns are often no longer usable, which greatly increases the processing cost.

[0018] Furthermore, protein layers or protein-DNA and protein-RNA layers lead to clogging of the pores, which increases the system pressure at a constant flow rate. A further increase in flow is accompanied by a further increase in pressure. This can lead to interruptions in the treatment. These apheresis columns are also no longer suitable for further use.

[0019] Typically, in affinity chromatography, the column used is first conditioned with a binding buffer that positively influences and promotes the interaction between the target substance and the ligand on the matrix substrate material. After the sample is applied and passed through the column, the bound target substance is flushed out or eluted. To elute the target molecule, an elution buffer specifically based on the binding mechanism is used to disrupt the interaction between the target substance and the ligand, such as by a competing ligand, by changing the pH, ionic strength or polarity. After the elution is complete, the column is washed again with a wash buffer or binding buffer. Elution is specific to the type of affinity chromatography, as it is used to dissolve a specific interaction between the target substance and the ligand.

[0020] For the selective removal of pathogenic substances from blood or plasma, prior art therapeutic apheresis is based on the principle of affinity chromatography, since this type of separation is based on the specific reversible interaction of a substance, such as a protein or a group of proteins, and a specific ligand coupled to a carrier matrix. The advantage of affinity chromatography purification is its high selectivity, which allows the targeted removal of the target substance from blood or plasma during therapeutic apheresis, while other components of the plasma are not or are only minimally removed due to nonspecific interactions. Affinity chromatography thus purposefully exploits the biological functions of the biomolecules or their individual structures. The interaction between the ligand and the target molecule can be the result of electrostatic or hydrophobic interactions, van der Waals forces or hydrogen bonds.

[0021] German patent application DE 100 65 241 A1 discloses an immunosorbent for extracorporeal apheresis and a method for preparing the same. The disclosed immunosorbent is intended to remove immunologically active substances, such as immunoglobulins, antibodies and immune complexes, from the plasma or whole blood of patients with immune diseases. The disclosed immunosorbent is prepared by covalently binding ligand protein A to different carrier materials.

[0022] It is known from the prior art that protein A shows a high affinity for the Fc region of polyclonal and monoclonal IgG antibodies. Therefore, a chromatographic column consisting of protein A bound to a matrix substrate material is suitable for affinity chromatography purification of antibodies, i.e. the chromatographic column is specific for antibodies but not for CRP. Therefore, DE 100 65 241 A1 does not disclose a single sampling column for the removal of CRP by affinity chromatography.

[0023] In DE 100 65 241 A1, for the adsorption of hIgG (Example 1D) onto a protein A adsorber, a binding buffer pH 7.0 and an elution buffer pH 2.2 were used in a standard cycle (single sampling elution buffer: 0.03-0.05 M citrate, 0.15 M NaCl). The disclosed protein A adsorber was tested for its chemical stability to various regeneration media in the pH range of 2-14 (Example 13D). As a regeneration medium, in particular, a 0.1 M sodium hydroxide solution (pH-value 14) was used. For regeneration, the respective regeneration medium was pumped through the column and then washed with a binding buffer at pH 7.0. Subsequently, the hIgG binding capacity was determined. The determination of the hIgG binding capacity was carried out here according to a standard cycle using a binding buffer pH 7.0 and an elution buffer pH 2.2. In addition, instead of human plasma, a hγ-globulin solution dissolved in a single sampling binding buffer at pH 7.0 (concentration 8 mg / mL) was used. The regeneration disclosed in DE 100 65 241 A1 is carried out in a separate and additional washing step, wherein after loading with hIgG the protein A adsorber is already treated with an elution buffer and a washing buffer at pH 2.2 before regeneration.

[0024] For therapeutic apheresis, additional rinsing steps for regenerating or also for washing the apheresis column represent a disadvantageous extension of the regeneration time and thus of the total treatment time. The more rinsing steps there are, the more regeneration solutions or washing solutions the regeneration cycle comprises, and the longer the total regeneration time of the apheresis column. In particular, if two adsorbers are used for a treatment, the plasma is pumped into one adsorber while the second adsorber is regenerated, it is not appropriate to increase the rinsing steps and use multiple regeneration solutions, since the regeneration of the second adsorber should be completed before the first adsorber is saturated in order to ensure a continuous and smooth removal of pathogenic substances from the blood or plasma.

[0025] Furthermore, DE 100 65 241 A1 only tests a single treatment with the respective regeneration medium. However, for therapeutic apheresis, the regeneration is reproducible, i.e. the regeneration medium can be used repeatedly without showing an adverse effect on the binding capacity, e.g. a reduction in the binding capacity for the substance to be removed from the blood or plasma. In therapeutic apheresis, the apheresis column is usually regenerated several times during a single treatment course.

[0026] Regarding repeated use, DE 100 65 241 A1 (Example 14D) describes the relevance of hIgG binding capacity to repeated use of 0.5N NaOH in an additional regeneration step. Therein, hIgG binding was performed using human plasma (4.2mL / mL gel; 6.5-fold excess of hIgG; concentration 9.8mg / mL) according to a standard cycle using a binding buffer pH 7.0 and an elution buffer pH 2.2. In the repeated regeneration after the standard cycle, 0.5N NaOH was pumped through the chromatographic column at a flow rate of 2mL / min for 2 hours (equivalent to 80 times the gel capacity). It was subsequently washed with water, a single sampling buffer of pH 2.2, and a single sampling buffer of pH 7.0. It is disclosed that when 0.5N NaOH was used, the hIgG binding capacity had dropped to 50% after 5 cycles. In another experiment, it is disclosed that when 0.5N NaOH was regenerated for 7 minutes in a 62.5mL adsorber, only 83% of the initial binding capacity was observed after 6 regeneration cycles. DE 100 65 241 A1 therefore teaches that sodium hydroxide solution can be used only to a limited extent as a regeneration medium, since a reduction in the binding capacity occurs in the case of repeated regenerations.

[0027] European patent application EP 3 459 552 A1 discloses the provision of a "universal" plasma, which is a mixture of plasma from different donors of different blood groups A, B, AB and / or O. The "universal" plasma has low levels of anti-A and anti-B antibodies and is therefore intended to be compatible with all blood groups. Anti-A antibodies and anti-B antibodies are thereby removed from the plasma mixture by purification by immunoaffinity chromatography. The matrix has oligosaccharide groups that are equivalent to epitopes of blood groups A and / or B. For regeneration, there is no further explanation of the treatment with sodium hydroxide, such as 1M NaOH.

[0028] The binding mechanism on which the immunoaffinity chromatography purification using a functionalized matrix is ​​based according to EP 3 459 552 A1 involves a specific carbohydrate-protein interaction. Therefore, EP 3 459 552 A1 does not disclose a single sampling column for the removal of CRP by affinity chromatography, in particular a single sampling column for the in vitro removal of CRP from blood or plasma and thus a single sampling column that can be used for therapeutic apheresis. In addition, EP 3 459 552 A1 also relates more to the field of preparative plasmapheresis, in which plasma is obtained during donation. This is different from therapeutic apheresis, in which the treated plasma is returned directly to the patient.

[0029] DE 100 65 241 A1 and EP 3 459 552 A1 do not disclose an apheresis column for the removal of CRP by affinity chromatography. With regard to apheresis columns for the removal of CRP by affinity chromatography, apart from the use of glycine / HCl or EDTA solutions for regeneration, no other compounds and methods are known in the prior art that allow the separation of molecules to be removed (in particular CRP) bound to the carrier material of the apheresis column. Therefore, there is a need for uses, methods and devices that can prevent the masking of binding sites and the increase of apheresis processing time and in particular ensure smooth flow.

[0030] It is therefore an object of the present invention to provide uses, methods and devices which are able to prevent masking of binding sites and an increase in apheresis processing time and in particular ensure a smooth flow.

[0031] Another object of the present invention is to provide a use, method and device for simplified regeneration of a single sampling column, in particular a single sampling column for the removal of CRP by affinity chromatography, whereby the disadvantages of the devices and methods known in the prior art are minimized. In other words, the object of the present invention is to provide a device for simplified regeneration of a single sampling column, in particular a simplified regeneration device for a single sampling column for the removal of CRP by affinity chromatography, which device can be operated with reduced training workload and thereby reduces the workload of personnel and reduces overall costs.

[0032] This object is solved by the teaching of the independent claims. Further advantageous designs are given by the description, the examples and the dependent claims. Summary of the invention

[0033] As used herein, the term "CRP" is equivalent to "C-reactive protein". Here, it preferably refers to human C-reactive protein. C-reactive protein (CRP) is a pentamer, each of which has two Ca 2+ Ion binding, with the help of which they can bind to ligands such as phosphorylcholine or its derivatives.

[0034] As used in this application, the term "affinity chromatography" with respect to the removal of CRP means that the removal of CRP occurs by specific binding between CRP and the components of the apheresis column used to remove CRP. In this case, one can also speak of "selective removal of CRP" or "selective CRP apheresis". This specific binding between CRP and the components of the apheresis column is based on the structural properties of the CRP protein, including, for example, the characteristic binding of CRP to phosphorylcholine and its derivatives or the binding of CRP to antibodies against CRP epitopes. The selective or molecule-specific removal of CRP includes the binding of CRP to the matrix in the apheresis column with a higher affinity than to other structures / molecules. In addition, CRP also binds to the matrix in the apheresis column with a higher affinity than other substances present in the blood, i.e., the matrix has specificity for CRP or the matrix is ​​specific for CRP. The matrix, preferably a solid phase modified with phosphorylcholine, preferably selectively binds CRP, i.e., almost exclusively binds CRP, without other blood components, such as LDL-cholesterol, antibodies or uremic toxins. Therefore, as disclosed herein, "removal of CRP" preferably means selective removal of CRP. However, the term "selective" as used in the present application for removing CRP does not mean that only CRP is removed. It is obvious to those skilled in the art that in the case of such affinity chromatography to remove CRP, other substances may inevitably (unintentionally) bind to the column material to a certain extent and are therefore also removed to a certain extent. An example is an antibody to phosphorylcholine, so it can also bind to the column material functionalized with phosphorylcholine. Another possibility is the non-specific binding of biological fluid components to, for example, matrix substrate materials, which can never be completely avoided.

[0035] As used herein, regeneration refers to the process by which a matrix of an apheresis column including a matrix substrate material and a ligand removes substances deposited thereon and returns to (ie, regenerates) a therapeutically usable state.

[0036] As used herein, in the affinity chromatography method for removing substances from a sample (herein, a biological liquid such as blood or plasma), the term "regeneration solution" refers to a solution loaded after the sample is loaded on the column material and the substance to be removed is specifically bound to the column material, so as to release the specific binding again, and thus release (or elute) the substance to be removed from the column material again. In addition to the regeneration solution, as used herein, in the affinity chromatography method for removing substances from a sample (here biological liquid, such as blood or plasma) (here selective removal of CRP), the term "elution buffer" (also referred to as "elution solution") refers to a solution loaded after the sample is loaded on the column material and the substance to be removed is specifically bound to the column material, so as to release the specific binding again, and thus release (or elute) the substance to be removed from the column material again. In contrast to the binding buffer, the conditions that do not promote the removal of substances and on the contrary prevent binding are created in the column material by the elution buffer.

[0037] In contrast to the elution buffer, the regeneration solution according to the present invention also removes the deposited substances here so that the single sampling column returns to or regenerates to a therapeutically usable state. In other words, the regeneration solution is used to release the specific binding of the substance to be removed to the column material to release (or elute) the substance to be removed from the column material again, and to remove the deposited substances so that the single sampling column returns to or regenerates to a therapeutically usable state. Therefore, the regeneration solution is used for elution and regeneration at the same time. Therefore, it is preferred that when the regeneration solution is used, the substance to be removed (here CRP) is also eluted. Therefore, it is preferred that the regeneration solution is used to elute CRP from the single sampling column used to remove CRP by affinity chromatography, so that the single sampling column is restored to a therapeutically usable state again.

[0038] As used herein, in affinity chromatography methods for removing substances from a sample (herein a biological fluid such as blood or plasma), the term "binding buffer" (also referred to as "binding solution") refers to a solution that is added to the sample and then loaded onto a column material together with the sample to remove the substance. Appropriate conditions for specific binding of the substance to be removed on the column material are ensured by the binding buffer.

[0039] As used herein, the term "biological fluid" refers to aqueous solutions present in mammals and preferably humans, such as cerebrospinal fluid, peritoneal fluid, pleural fluid, ascites, blood, plasma, liver extracts and interstitial fluid. The present invention preferably relates to biological fluids containing CRP.

[0040] The use of glycine / HCl or EDTA solutions for the regeneration of apheresis columns, in particular CRP apheresis columns, is known from the prior art. By a sudden change in pH to the acidic range, bound proteins are denatured or the bonds between the adsorbed molecules to be removed (in this case, in particular CRP) and the support material of the apheresis column are released by complexation of mediated bound cations (in particular calcium). Similar to the addition of EDTA, citrate-containing solutions are suitable as elution buffers for releasing bound CRP from apheresis columns. In the prior art, no compounds are known that are capable of separating CRP to be removed bound to the support material of apheresis columns by other mechanisms.

[0041] In order to test the tolerance of CRP selective matrix substrate material described herein to glycine / HCl solution, the inventor used 7.5 matrix volumes (MV) of glycine / HCl buffer, followed by 4MV of PBS solution (pH 7.4) and then 4MV of NaCl, and carried out 200 regeneration cycles, wherein plasma was not loaded to the CRP-single sampling column between the regeneration cycles. Therefore, the CRP binding capacity was tested once before and after 200 consecutive regeneration cycles. It was shown that the repeated pH-changes from 7.4 to 2.8 had no effect on the CRP binding capacity of the CRP single sampling column used. Using glycine / HCl pH 2.8 as a regeneration solution would not reduce the CRP binding capacity due to the degradation of the column material or adsorber.

[0042] The removal of CRP was studied on a laboratory scale using a column containing a CRP selective matrix substrate material and human plasma loaded on the column. The chromatographic column used contained 0.5g of matrix and 75mL (150 matrix volumes) of human plasma containing 100, 50 or 10mg / L of CRP, circulated at 1.2mL / min (corresponding to 17mL / min single column scale). The matrix was washed with buffer (0.1M Tris, 0.2M NaCl and 2mM CaCl2), CRP was eluted with elution buffer (EDTA, 0.2M NaCl and 0.1MTris), and the column was regenerated with glycine / hydrochloric acid pH 2.8. It was found that the matrix used specifically and selectively binds to CRP. When EDTA elution buffer was used, calcium-dependent binding could be prevented and CRP was completely eluted. When it was regenerated with glycine / HCl, almost no other proteins were detected. No decrease in CRP binding capacity after multiple uses of the column was observed.

[0043] Clinical apheresis is performed in a cycle, including pre-rinsing the adsorber with 0.9% NaCl buffer, loading the adsorber with the plasma that will be returned to the patient, and regenerating the adsorber to prepare it for the next cycle. Except for plasma, no other solution used is returned to the patient. In order to study the removal of CRP in plasma under clinical conditions, 20mL of matrix substrate is loaded into the column. It is then pre-rinsed with 200mL of 0.9% NaCl buffer, and then 2.5L of human plasma with an initial concentration of 100mg / LCRP is passed through at a flow rate of 30mL / min in 5 cycles (1L / cycle). After each cycle, the apheresis column is regenerated with 35mL of 0.9% NaCl, 50mL of glycine / HCl buffer, 80mL of PBS and 35mL of 0.9% NaCl. The matrix used contains a phosphorylcholine ligand for human CRP from plasma, which is particularly suitable for therapeutic apheresis. Therefore, the matrix is ​​very selective in binding CRP, and only very low non-specific protein binding with other plasma proteins is observed. CRP binds to phosphorylcholine and its derivatives in a highly specific manner through electrostatic interactions between the ligand and CRP.

[0044] In the apheresis therapy for selectively removing CRP from plasma, an average of 6000 mL of plasma is processed in 6 to 12 cycles in one course of treatment. Including the regeneration of the apheresis column, it usually takes about 4-5 hours to process 6000 mL of plasma. Regeneration is performed using a regeneration solution glycine / HCl buffer known in the prior art. Compared with the initial concentration in the blood, the minimal loss of other plasma proteins caused by CRP apheresis can be attributed to the loading load and regeneration of the changes in the adsorber during treatment, and is less than other in vitro methods. The nonspecific binding of plasma proteins to the adsorber matrix is ​​negligible. It has been shown that patients can therefore receive CRP apheresis treatment without restriction because blood loss is minimal and no side effects have been reported so far.

[0045] In therapeutic apheresis to remove CRP from blood or plasma, the plasma is returned to the patient after passing through an apheresis column. Reusable adsorbers can only be used for the same patient. However, when using apheresis columns to remove CRP from circulating blood or plasma in therapeutic apheresis, the inventors of the present invention have found that during regeneration with glycine / HCl, a protein layer is formed around the matrix particles (e.g., agarose particles), which may lead to an increase in this effect, especially when the patient's blood to be purified contains high concentrations of cell-free DNA / RNA. The formation of a protein layer in the apheresis column masks the binding sites and reduces the performance of the apheresis material. Further regeneration attempts by known measures, such as with glycine / HCl solutions, could not restore the original state.

[0046] As damage to the apheresis column progresses, the patient's treatment time increases and the patient's pain also increases. In addition, damaged apheresis columns are often no longer usable, which greatly increases the cost of disposal. In addition, the protein layer or protein-DNA and protein-RNA layer leads to clogging of the pores, which increases the system pressure at a constant flow rate. A further increase in flow is accompanied by a further increase in pressure. This can lead to interruptions in treatment. These apheresis columns are also no longer suitable for further use.

[0047] In therapeutic apheresis, the substance to be removed (here CRP) is removed from the blood or plasma of an individual patient. The composition of the plasma is individualized for the patient concerned. In this regard, different patients' plasma naturally contains different concentrations of DNA / RNA free from cells. The use of glycine / HCl buffer to regenerate the CRP apheresis column will therefore lead to different degrees of the above-mentioned problems depending on the patient, which have been shown in the case of glycine / HCl buffer regeneration. Therefore, regardless of the concentration of DNA / RNA free from cells in the plasma, it will be particularly advantageous to use a regeneration solution that allows complete regeneration of the apheresis column. In addition, for regeneration, it will be particularly advantageous to use a regeneration solution that improves the reusability of the apheresis column.

[0048] For regeneration, it is also possible to consider using different regeneration solutions through the apheresis column one after another for the regeneration of the apheresis column. However, an increase in the number of flushing steps or an increase in the number of different regeneration solutions will also lead to an increase in the total duration of regeneration. Therefore, this will also affect the treatment time and increase the patient's suffering time. Although the flow rate during blood purification is limited by the patient's blood flow, it is in principle possible to increase the flow rate during regeneration to speed up the process and thus make the apheresis column available for further use. However, higher flow rates lead to a simultaneous increase in the pressure on the apheresis and the matrix or column material. Depending on the column material, above a certain pressure, the shape and strength of the material will change, which will reduce the separation performance of the column material, so that the flow rate for the regeneration of the apheresis column depends on the pressure resistance of the column material used. Large volume flow rates compress the matrix, which leads to a decrease in the flow rate. Therefore, regeneration at an arbitrary rate is not possible. However, if the volume flow rate is reduced, the compression is reversible. Therefore, the regeneration time depends mainly on the volume and flow rate of the individual flushing steps, and therefore also on the number of flushing steps.

[0049] It is therefore particularly advantageous to use a regeneration solution for regeneration which, when used alone, already enables complete regeneration of the apheresis column. By using a single regeneration solution for regenerating the apheresis column, the number of flushing steps can be reduced to the required minimum. In particular, it is advantageous if the regeneration solution is used to elute the substance to be removed (here CRP) and, at the same time, to restore the apheresis column to a therapeutically usable state.

[0050] Surprisingly, it has been found that already used apheresis columns, i.e. apheresis columns after treatment for removing substances from the blood, can be regenerated with the aid of alkali metal hydroxide solutions, preferably sodium hydroxide solutions, and that apheresis columns regenerated in this way can still be used for apheresis. In particular, apheresis columns are regenerated which, due to their use, have protein deposits and / or protein-DNA and protein-RNA deposits which can no longer be removed by the regeneration solutions used in the prior art and which have therefore become unusable.

[0051] Therefore, the present invention relates to the use of an alkali metal hydroxide solution (preferably a sodium hydroxide solution) to regenerate a single sampling column. In other words, the present invention is intended to use an alkali metal hydroxide solution (preferably a sodium hydroxide solution) as a regeneration solution in single sampling. In more other words, the present invention is intended to use an alkali metal hydroxide solution (preferably a sodium hydroxide solution) as a regeneration solution for a single sampling column.

[0052] More specifically, the present invention relates to the use of an alkali metal hydroxide solution (preferably a sodium hydroxide solution) to regenerate a single sampling column for affinity chromatography to remove CRP. Therefore, the present invention preferably relates to the use of an alkali metal hydroxide solution for regenerating a single sampling column, wherein the single sampling column is a single sampling column for affinity chromatography to remove CRP. The present invention particularly preferably relates to the use of an alkali metal hydroxide solution for regenerating a single sampling column, wherein the single sampling column is a single sampling column for affinity chromatography to remove CRP from blood or plasma.

[0053] Surprisingly, it has been found that alkaline regeneration with an alkali metal hydroxide solution, preferably a sodium hydroxide solution, can also regenerate already damaged adsorber matrices in a single sampling column. If only an alkali metal hydroxide solution, preferably a sodium hydroxide solution, is used as a regeneration means, no acidic protein precipitation occurs. This is an important aspect of restoring the function of a single sampling column. In particular, it has been shown that alkaline regeneration with an alkali metal hydroxide solution, preferably a sodium hydroxide solution, can regenerate already damaged adsorber matrices in a single sampling column for the removal of CRP by affinity chromatography.

[0054] Furthermore, it is advantageous that the single sampling column can be stored by using an alkali metal hydroxide solution (preferably a sodium hydroxide solution) as a storage agent. In a preferred embodiment, an alkali metal hydroxide solution (preferably a sodium hydroxide solution) can be used to store a single sampling column for affinity chromatography to remove CRP. This allows single sampling treatment to be performed as efficiently as possible, including preparations for the storage of the single sampling column, because time-consuming washing steps of the single sampling column after treatment can be avoided. Alkali metal hydroxide solutions are suitable for the storage of single sampling columns, and in particular sodium hydroxide solutions are very suitable for the storage of single sampling columns because their bactericidal effect is very good. The growth of microorganisms is substantially eliminated.

[0055] Therefore, another aspect of the present invention relates to the use of an alkali metal hydroxide solution (preferably a sodium hydroxide solution) for regenerating and storing a single sampling column (preferably a single sampling column for removing CRP by affinity chromatography). Therefore, the present invention is also intended to use an alkali metal hydroxide solution (preferably a sodium hydroxide solution) for regenerating and storing a single sampling column. In other words, the present invention is also intended to use an alkali metal hydroxide solution (preferably a sodium hydroxide solution) as a regeneration solution in a single sampling and for storing a single sampling column. More preferably, the present invention is intended to use an alkali metal hydroxide solution (preferably a sodium hydroxide solution) as a regeneration solution in a single sampling for removing CRP by affinity chromatography and for storing a single sampling column for removing CRP by affinity chromatography.

[0056] Compared to the methods known from the prior art for regenerating apheresis columns, the alkali metal hydroxide solution (preferably sodium hydroxide solution) is characterized in that it is not a chelating agent for cations. It is assumed that the alkali metal hydroxide (preferably sodium hydroxide) on the one hand reduces the binding affinity of the substance to be released to the carrier material and on the other hand denatures the proteins that may bind and thereby reduces the binding to the carrier material. Rapid regeneration allows the chromatographic column to be restored to a usable state during the apheresis treatment. The regeneration of the apheresis column can be performed during the apheresis treatment, but has no effect on the apheresis treatment itself and is therefore not a diagnostic or therapeutic method, but a method for purifying the apheresis column, which does not involve an apheresis patient, even if the regeneration of the apheresis column occurs during the apheresis treatment.

[0057] In the use of an alkali metal hydroxide solution (preferably a sodium hydroxide solution) according to the present invention as described herein for regenerating a single sampling column (preferably a single sampling column for removing CRP by affinity chromatography), the regeneration of the single sampling column can be performed during the single sampling treatment. Preferably, in the use of an alkali metal hydroxide solution (preferably a sodium hydroxide solution) according to the present invention as described herein for regeneration of a single sampling column, regeneration of the single sampling column occurs during the removal of target compounds (particularly CRP). Therefore, a preferred embodiment of the present invention is to use an alkali metal hydroxide solution (preferably a sodium hydroxide solution) for the regeneration of a single sampling column, wherein regeneration occurs during the single sampling treatment.

[0058] Therefore, the present invention also relates to the use of an alkali metal hydroxide solution for regenerating a single sampling column, wherein the single sampling column is a single sampling column for removing CRP by affinity chromatography, wherein the regeneration is completed during the single sampling treatment. Therefore, the present invention also relates to the use of an alkali metal hydroxide solution for regenerating a single sampling column, wherein the single sampling column is a single sampling column for removing CRP by affinity chromatography, wherein the regeneration is completed during the single sampling treatment for removing CRP from blood or plasma in vitro. During the regeneration process, the single sampling column is separated from the extracorporeal circulation system. In other words, the single sampling column is not connected to the extracorporeal circulation system during the regeneration process. In other words, the single sampling column is not fluidically connected to the extracorporeal circulation system during regeneration.

[0059] Therefore, the present invention also relates to the use of an alkali metal hydroxide solution for regenerating an apheresis column, wherein the apheresis column is an apheresis column for removing CRP by affinity chromatography, wherein the regeneration is performed during an apheresis treatment for the extracorporeal removal of CRP from blood or plasma, wherein the apheresis column is separated from the extracorporeal circulation system during the regeneration process.

[0060] Therefore, the present invention preferably relates to the use of an alkali metal hydroxide for regenerating an apheresis column, wherein the apheresis column is an apheresis column for the removal of CRP by affinity chromatography, wherein the regeneration takes place during an apheresis treatment for the extracorporeal removal of CRP from blood or plasma, wherein the apheresis column is not connected to an extracorporeal circulation system during the regeneration process.

[0061] Preferably, the use of an alkali metal hydroxide solution for regenerating an apheresis column is an apheresis column for removing CRP by affinity chromatography when plasma has passed through an apheresis column for removing CRP before regeneration.

[0062] Therefore, it is preferred to use an alkali metal hydroxide solution to regenerate the single sampling column, wherein the single sampling column is a single sampling column for removing CRP by affinity chromatography, wherein the single sampling column contains CRP. It is also preferred to use an alkali metal hydroxide solution for regenerating a single sampling column, wherein the single sampling column is a single sampling column for removing CRP by affinity chromatography, wherein the single sampling column is saturated with CRP.

[0063] "During apheresis treatment" or "during the in vitro removal of a target compound (particularly CRP in the blood)" refers herein to the regeneration of the apheresis column being completed during a therapeutic operation in the same apheresis device. Thus, the in vitro removal of the target compound (particularly CRP in the blood) can be interrupted, and the same device can be used to regenerate the apheresis column. Alternatively, regeneration of the apheresis column can be performed while the in vitro removal of the target compound (particularly CRP in the blood) continues by another apheresis column. Regeneration of the apheresis column is completed in the apheresis device. However, the term "during apheresis treatment" does not mean that the regeneration and removal of CRP from the blood occur simultaneously on the same apheresis column.

[0064] Furthermore, as already mentioned, the inventors have recognized that deposits form in the apheresis column during regeneration with glycine / HCl (protein layer). The effect can be enhanced if the patient's blood to be purified contains high concentrations of cell-free DNA / RNA. The formation of deposits in the apheresis column leads to an irreversible sticking of the matrix, i.e. no further regeneration attempts can be made by known measures to restore the conditions, such as with glycine / HCl solution. In the event that proteins are deposited on the column material of the apheresis column until a protein layer is formed on the column material of the apheresis column, it is no longer possible to regenerate the apheresis column by known measures, such as flushing with glycine / HCl solution, because the protein deposits or protein layer can no longer be removed with conventional regeneration solutions.

[0065] Surprisingly, it was found that 0.01M to 1M alkali metal hydroxide solution, preferably 0.04M to 0.4M alkali metal hydroxide solution, allows protein deposits to be removed again, preferably completely. It was surprisingly found that in particular 0.01M to 1M sodium hydroxide solution, preferably 0.04M to 0.4M sodium hydroxide solution, allows protein deposits to be removed again, preferably completely. The relevance of alkaline regeneration solutions, for example in the pH range of 12-14, to the regeneration of CRP apheresis columns is not known in the prior art.

[0066] Therefore, the use according to the present invention preferably refers to an apheresis column containing DNA / RNA (free DNA or free RNA), preferably refers to an apheresis column on which there is DNA or RNA bound to a matrix, and particularly preferably refers to an apheresis column on which the DNA or RNA is bound to a ligand of the matrix.

[0067] Therefore, a preferred embodiment of the present invention relates to the use of an alkali metal hydroxide solution, preferably a sodium hydroxide solution, to regenerate an apheresis column, preferably an apheresis column for the removal of CRP by affinity chromatography, wherein the apheresis column contains a DNA and / or RNA deposit, i.e. on an adsorber or on an adsorber matrix as also referred to herein.

[0068] Therefore, the present invention also relates to the use of an alkali metal hydroxide solution for regenerating an apheresis column, wherein the apheresis column is an apheresis column for the removal of CRP by affinity chromatography, wherein the regeneration is accomplished during an apheresis treatment for the extracorporeal removal of CRP from blood or plasma, and wherein the apheresis column is separated from the extracorporeal circulation system during the regeneration, wherein the apheresis column comprises a DNA and / or RNA deposit, i.e. on an adsorber or as described herein on an adsorber matrix.

[0069] As used herein, "free DNA" or "free RNA" refers to deoxyribonucleic acid or ribonucleic acid, respectively, that is located outside of a cell. DNA / RNA bound to a single sampling column ligand is also referred to as free DNA / free RNA because it is also located outside of a cell.

[0070] Surprisingly, using alkali metal hydroxide solution to regenerate the single mining column, preferably for affinity chromatography to remove CRP, in addition to restoring the single mining column to a treatment-usable state, it is also allowed to simultaneously elute the bound CRP. Therefore, it is preferred to use alkali metal hydroxide solution to regenerate the single mining column, elute the bound CRP, so that the single mining column is restored to a treatment-usable state. It is particularly preferred to use alkali metal hydroxide solution to regenerate the single mining column, wherein the single mining column is a single mining column for affinity chromatography to remove CRP, wherein the CRP bound to the single mining column is eluted by alkali metal hydroxide solution. Therefore, it is particularly preferred to use alkali metal hydroxide solution to regenerate the single mining column, wherein the single mining column is a single mining column for affinity chromatography to remove CRP, wherein the bound CRP is eluted and the single mining column is brought back to a treatment-usable state at the same time.

[0071] The use of an alkali metal hydroxide solution to regenerate a single sampling column, preferably a single sampling column for affinity chromatography to remove CRP, and the possibility of simultaneously eluting bound CRP, can thereby advantageously dispense with the use of other regeneration solutions than the alkali metal hydroxide solution according to the invention. Thus, the use of an alkali metal hydroxide solution to regenerate a single sampling column eliminates the need for additional regeneration with glycine / HCl or EDTA to elute bound CRP from the CRP single sampling column.

[0072] In a preferred embodiment, no additional regeneration solution is used for regeneration in addition to the alkali metal hydroxide solution used for regeneration of the single sampling column. In a preferred embodiment, no additional elution buffer is used to elute the CRP in addition to the alkali metal hydroxide solution used for regeneration of the single sampling column. Therefore, the use of an alkali metal hydroxide solution to regenerate a single sampling column, preferably a single sampling column for affinity chromatography to remove CRP, is particularly advantageous because the number of flushing steps for regeneration of the single sampling column does not have to be increased and the total regeneration time is not extended.

[0073] Therefore, preferably alkali metal hydroxide solution is used for the purpose of single acquisition of the regeneration of column, wherein the single acquisition column is a single acquisition column for affinity chromatography to remove CRP, wherein other regeneration solutions are not used. Therefore, preferably alkali metal hydroxide solution is used for the purpose of single acquisition of the regeneration of column, wherein the single acquisition column is a single acquisition column for affinity chromatography to remove CRP, wherein glycine / HCl is not used for elution or regeneration. Therefore, preferably alkali metal hydroxide solution is used for the purpose of single acquisition of the regeneration of column, wherein the single acquisition column is a single acquisition column for affinity chromatography to remove CRP, wherein elution buffer or other regeneration solutions for eluting the bound CRP are not used before using alkali metal hydroxide solution. However, it is not excluded that flushing solution or neutralization solution, such as NaCl solution or PBS solution, are used before and after this.

[0074] Regeneration of the apheresis column can be performed during apheresis treatment. In this case, plasma loss can be minimized by removing plasma from parts of the apheresis device, in particular from the apheresis column, before introducing the alkali metal hydroxide solution (preferably sodium hydroxide solution), for which purpose a flushing solution can be used. The alkali metal hydroxide solution (preferably sodium hydroxide solution) can be further removed from the apheresis column by a neutralization solution in preparation for apheresis treatment.

[0075] Therefore, preference is given to the use of an alkali metal hydroxide solution for the regeneration of an apheresis column, wherein the apheresis column is an apheresis column for the removal of CRP by affinity chromatography, wherein a rinse solution is introduced before the introduction of the alkali metal hydroxide solution, and wherein the rinse solution is introduced after the introduction of the alkali metal hydroxide solution, or a neutralization solution is introduced followed by the rinse solution.

[0076] Therefore, the use of alkali metal hydroxide solution for the regeneration of the single sampling column is preferred, wherein the single sampling column is a single sampling column for the removal of CRP by affinity chromatography, wherein a flushing solution is introduced into the single sampling column before the introduction of the alkali metal hydroxide solution. The flushing solution is preferably a sodium chloride solution, in particular a physiological sodium chloride solution or a PBS solution (phosphate buffered saline). Preferably, a NaCl solution is used, particularly preferably a physiological NaCl solution as the flushing solution.

[0077] Therefore, the use of alkali metal hydroxide solution for the regeneration of the single sampling column is preferred, wherein the single sampling column is a single sampling column for the removal of CRP by affinity chromatography, wherein the rinsing solution is introduced after the introduction of the alkali metal hydroxide solution. The rinsing solution is preferably a sodium chloride solution, in particular a physiological sodium chloride solution or a PBS solution (phosphate buffered saline). Preferably, a NaCl solution is used, particularly preferably a physiological NaCl solution as the rinsing solution.

[0078] Therefore, preferably alkali metal hydroxide solution is used for the purpose of regeneration of single mining column, wherein single mining column is used for affinity chromatography to remove CRP single mining column, wherein after introducing alkali metal hydroxide solution, neutralizing solution is introduced, and then flushing solution is introduced. Flushing solution is preferably sodium chloride solution, particularly physiological sodium chloride solution or PBS solution (phosphate buffered saline). Flushing solution is preferably NaCl solution, particularly preferably physiological NaCl solution. Neutralizing solution is preferably selected from the group comprising the following or consisting of the following: PBS solution or NaCl solution or citrate solution. Particularly preferably, the neutralizing solution is a citrate solution.

[0079] Particularly preferably, the flushing solution is not an elution buffer for removing bound CRP. Particularly preferably, the flushing solution is even more not a regeneration solution. In other words, it is particularly preferred that the flushing solution is not a glycine / HCl buffer. It is further preferred that no regeneration solution other than an alkali metal hydroxide solution is used to regenerate the single sampling column. Therefore, it is preferred that no other regeneration solution is introduced before the introduction of the flushing solution and after the introduction of the alkali metal hydroxide solution and after the subsequent introduction of the flushing solution or the neutralization solution and the flushing solution.

[0080] Therapeutic apheresis is performed in cycles, alternating between loading and regeneration of the apheresis column. Preferably, an alkali metal hydroxide solution is used as the regeneration solution in each regeneration cycle. Further preferably, an alkali metal hydroxide solution is used as the only regeneration solution.

[0081] Therefore, it is preferred to repeatedly use an alkali metal hydroxide solution to regenerate the single sampling column, wherein the single sampling column is a single sampling column for removing CRP by affinity chromatography. Therefore, it is preferred to use an alkali metal hydroxide solution to regenerate the single sampling column, wherein the single sampling column is a single sampling column for removing CRP by affinity chromatography, wherein the alkali metal hydroxide solution is repeatedly used to regenerate the single sampling column. Therefore, it is preferred to use an alkali metal hydroxide solution to regenerate the single sampling column, wherein the single sampling column is a single sampling column for removing CRP by affinity chromatography, wherein the alkali metal hydroxide solution is used to regenerate the single sampling column in each regeneration cycle.

[0082] The advantage of using an alkali metal hydroxide solution, preferably a sodium hydroxide solution, for regenerating a single sampling column, preferably a single sampling column for affinity chromatography to remove CRP, is that the number of flushing steps for regenerating the single sampling column does not have to be increased compared to the prior art using a glycine / HCl buffer using an alkali metal hydroxide solution, because the alkali metal hydroxide solution can be advantageously used to elute the bound CRP and simultaneously to remove other substances. Another advantage is that when regenerating a single sampling column using an alkali metal hydroxide solution, acidic protein folding no longer occurs, and the problems shown when regenerating a single sampling column using a glycine / HCl buffer do not occur. Therefore, regenerating a single sampling column using an alkali metal hydroxide solution significantly improves the reusability of the single sampling column. It has been shown that a CRP adsorber can be regenerated with a sodium hydroxide solution for 200 cycles without a reduction in performance.

[0083] Therefore, it is preferred to use an alkali metal hydroxide solution to regenerate the single sampling column, wherein the single sampling column is a single sampling column for removing CRP by affinity chromatography, wherein the single sampling column can be reused at least 50 times, preferably at least 100 times, more preferably at least 150 times, and most preferably at least 200 times.

[0084] regeneration solution

[0085] Glycine-HCl solutions used as regeneration solutions are known in the prior art. The pH value of the glycine / HCl solution is in the range of 2-3, for example pH=2.8. The concentration range of the glycine-HCl solution is 0.1M to 1M. For the preparation of the glycine-HCl solution or a synonymous solution using a glycine-HCl buffer, 15.01 g of glycine is dissolved in 1 liter of water and 25% hydrochloric acid is added. The pH value can be adjusted using hydrochloric acid or sodium hydroxide. The preparation of the glycine-HCl solution is known to those skilled in the art.

[0086] "Sodium hydroxide solution" refers herein to a solution comprising sodium hydroxide in a solvent such as water or an alcohol such as methanol, ethanol, propanol, etc. or in a solvent mixture of water and at least one alcohol. Preferably, it comprises or consists of sodium hydroxide in water. Thus, aqueous sodium hydroxide solution (also referred to as NaOH aq ).

[0087] Sodium hydroxide can be obtained in various purities, preferably 98%, more preferably 99%, more preferably 99.5%, more preferably 99.9%, and most preferably 99.99%. Preferably, the purity is between 98% and 100%.

[0088] In principle, the sodium hydroxide in the sodium hydroxide solution can exist in all available concentrations (based on the sodium hydroxide in the solvent or solvent mixture). Usually, the solution is an aqueous sodium hydroxide solution. However, the concentration of sodium hydroxide in the preferred sodium hydroxide solution is in the range of 0.005mol / l to 1.0mol / l, more preferably between 0.01mol / l to 1.0mol / l, further preferably between 0.02mol / l to 0.80mol / l. Further preferably between 0.03mol / l to 0.60mol / l, further preferably between 0.04mol / l to 0.50mol / l, further preferably between 0.05mol / l to 0.40mol / l, further preferably between 0.06mol / l to 0.30mol / l, further preferably between 0.07mol / l to 0.20mol / l, further preferably between 0.08mol / l to 0.10mol / l.

[0089] A preferred embodiment of the present invention relates to the use of an alkali metal hydroxide solution (preferably a sodium hydroxide solution) for the regeneration of a single sampling column, wherein the concentration of the alkali metal hydroxide, preferably sodium hydroxide, in the solution is in the range of 0.01-1.0 mol / l.

[0090] Another preferred embodiment of the present invention relates to the use of an alkali metal hydroxide solution (preferably a sodium hydroxide solution) for the regeneration of a single sampling column, wherein the concentration of the alkali metal hydroxide (preferably sodium hydroxide) in the solution is in the range of 0.05-0.20 mol / l, preferably 0.05-0.10 mol / l, more preferably 0.07-0.10 mol / l.

[0091] The alkaline pH value of the alkali metal hydroxide solution (preferably sodium hydroxide solution) is not limited. Preferably, the alkali metal hydroxide solution (preferably sodium hydroxide solution) has a pH of 7 to 14, more preferably 7.5 to 14, more preferably 8.0 to 14.0, more preferably 8.5 to 14, more preferably 9.0 to 14.0, more preferably 9.5 to 14.0, more preferably 10.5 to 14.0, more preferably 11 to 14.0, more preferably 11.5 to 14.0, more preferably 12 to 14.0, more preferably 12.5 to 14.0, even more preferably 13 to 14. Particularly preferably, the sodium hydroxide solution has a pH of 12 to 13.7.

[0092] Therefore, one embodiment of the present invention is to use an alkali metal hydroxide solution (preferably a sodium hydroxide solution) for the regeneration of the single sampling column, wherein the alkali metal hydroxide solution (preferably a sodium hydroxide solution) has a pH value in the range of 12 to 14.

[0093] It is also preferred to use an alkali metal hydroxide solution for the regeneration of the single sampling column, wherein the single sampling column is a single sampling column for the removal of CRP by affinity chromatography, wherein the alkali metal hydroxide solution (preferably sodium hydroxide solution) has a pH value in the range of 12 to 14, more preferably a pH value of 12 to 13.7.

[0094] As used herein, the term "alkali hydroxide solution" or "alkali metal hydroxide solution" refers to "lithium hydroxide solution", "sodium hydroxide solution", "potassium hydroxide solution" or a mixture of two or three of the above solutions.

[0095] "Lithium hydroxide solution" refers herein to a solution comprising lithium hydroxide in a solvent such as water or an alcohol such as methanol, ethanol, propanol, etc., or in a solvent mixture of water and at least one alcohol. Preferably, it comprises or consists of a solution of lithium hydroxide in water.

[0096] "Potassium hydroxide solution" herein refers to a solution comprising potassium hydroxide in a solvent such as water or an alcohol such as methanol, ethanol, propanol, etc. or in a solvent mixture of water and at least one alcohol. Preferably, it comprises or consists of a solution of potassium hydroxide in water.

[0097] In principle, the lithium hydroxide in the lithium hydroxide solution or the potassium hydroxide in the potassium hydroxide solution can be present in all useful concentrations (based on the lithium hydroxide in the solvent or solvent mixture or based on the potassium hydroxide in the solvent or solvent mixture). However, the concentration of the lithium hydroxide in the lithium hydroxide solution or the potassium hydroxide in the potassium hydroxide solution is preferably 0.005 to 1.0 mol / l, more preferably 0.01 to 1.0 mol / l, more preferably 0.02 to 0.80 mol / l, still more preferably 0.03 to 0.60 mol / l, further preferably 0.04 to 0.50 mol / l, further preferably 0.05 to 0.40 mol / l, further preferably 0.06 to 0.30 mol / l, further preferably 0.07 to 0.20 mol / l, further preferably 0.08 to 0.10 mol / l.

[0098] If two of the above three alkali metal hydroxide solutions are used, the cumulative concentration of the two alkali metal hydroxide solutions should be in the range of 0.005 mol / l to 1.0 mol / l, more preferably between 0.01 mol / l and 1.0 mol / l, further preferably between 0.02 mol / l and 0.80 mol / l, further preferably between 0.03 mol / l and 0.60 mol / l, further preferably between 0.04 mol / l and 0.50 mol / l, further preferably between 0.05 mol / l and 0.40 mol / l, further preferably between 0.06 mol / l and 0.30 mol / l, further preferably between 0.07 mol / l and 0.20 mol / l, further preferably between 0.08 mol / l and 0.10 mol / l. This means that, for example, a potassium hydroxide concentration of, for example, 0.06 mol / l in a solution and a sodium hydroxide concentration of, for example, 0.08 mol / l in the same solution results in a cumulative alkali hydroxide concentration of 0.14 mol / l of solution. The same applies if all three of the abovementioned alkali hydroxides are used in one solution, for example 0.04 mol / l of lithium hydroxide, 0.02 mol / l of potassium hydroxide and 0.05 mol / l of sodium hydroxide, so that the cumulative alkali hydroxide concentration in the solution is 0.11 mol / l.

[0099] Therefore, one embodiment of the present invention relates to the use of an alkali metal hydroxide solution selected from the group comprising or consisting of a sodium hydroxide solution, a lithium hydroxide solution and / or a potassium hydroxide solution for the regeneration of the single sampling column.

[0100] However, it is particularly preferred to use sodium hydroxide solution to regenerate the single sampling column.

[0101] Furthermore, alkali metal hydroxide solutions are able to regenerate adsorbers which have formed protein deposits which can no longer be removed by conventional regeneration methods.

[0102] Therefore, preference is given to the use of an alkali metal hydroxide solution for the regeneration of an apheresis column, wherein the apheresis column is a apheresis column for the removal of CRP by affinity chromatography, wherein the regeneration is done when protein deposits can no longer be removed by conventional regeneration methods. Therefore, preference is given to the use of an alkali metal hydroxide solution for the regeneration of an apheresis column, wherein the apheresis column is a apheresis column for the removal of CRP by affinity chromatography, wherein the regeneration is done when protein deposits can no longer be removed by regeneration agents such as glycine-HCl or EDTA.

[0103] Single mining column

[0104] How to design or construct apheresis columns (or cassettes or cartridges) in principle is part of the prior art and can be learned in particular from EP 0237659 B1. The exact dimensions of the cassette, column or cartridge used according to the invention (as a device for selectively removing target compounds, in particular CRPs) depend largely on the intended use of the device according to the invention. Apheresis columns generally comprise a housing, for example in the form of a cassette or cartridge, which is fluidically connected to an extracorporeal circulation system via at least one inlet and at least one outlet and contains a matrix for affinity chromatography or adsorption removal of target compounds, in particular CRPs.

[0105] "Target compound" herein refers to a substance removed from blood by apheresis, ie a substance that should be bound to a matrix during apheresis. Particularly preferably, the target compound removed from blood or plasma by apheresis is CRP herein.

[0106] The matrix for affinity chromatography (or adsorption) removal of the target compound (especially CRP) comprises a matrix substrate material (column material), on which a compound (ligand) having the property of specifically binding the target compound, especially CRP, is again bound. According to a preferred embodiment of the present invention, the matrix is ​​integrated into or fixed in a single sampling column for affinity chromatography to remove the target compound, especially CRP, so that it does not flow out of the column with the plasma flow. This depends on the implementation form, for example, in the form of filters at the inlet and outlet of the device.

[0107] The use according to the invention is particularly useful in regenerating an apheresis column for removing CRP by affinity chromatography.

[0108] In principle, all inert chromatographic or column materials are suitable as matrix substrate materials (column materials) for producing the matrix, which in particular do not react with blood or plasma or change or contaminate blood or plasma in such a way that blood or plasma cannot be returned to the patient after contact with the matrix. Therefore, suitable matrix substrate materials according to the invention include but are not limited to Polyvinyl pyrrolidone, methacrylate, methacrylate resin, agarose, cross-linked agarose such as Preferably, the matrix substrate material is selected from the group comprising or consisting of the following: agarose and sepharose. Particularly preferably, the matrix substrate material is agarose. The solid phase silica matrix can actually comprise almost any form of granular silicon dioxide, including amorphous silicon dioxide, such as colloidal silicon dioxide, silica gel, precipitated silicon dioxide and fumed or pyrogenic silicon dioxide; microcrystalline silicon dioxide such as diatomaceous earth; and crystalline silicon dioxide such as quartz.

[0109] Although the flow rate during blood purification is limited by the patient's blood flow, it is in principle possible to increase the flow rate during regeneration to speed up the process and thus make the apheresis column available for further use. However, a higher flow rate leads to a simultaneous increase in the pressure on the apheresis column and thus to an increase in the pressure on the matrix or column material. Depending on the column material, above a certain pressure the shape and strength of the material changes and thereby reduces the separation performance of the column material, so that the flow rate for regenerating an apheresis column depends on the pressure resistance of the column material used.

[0110] It has been found herein that the use of an alkali metal hydroxide solution, preferably a sodium hydroxide solution, not only allows for higher pressures, but even such high pressures can be used that the single sampling column can be purified in only minutes or seconds. In particular, the combination of an alkali metal hydroxide solution, preferably a sodium hydroxide solution, and agarose and its derivatives is suitable for rapid regeneration. Therefore, one embodiment of the present invention relates to the use of an alkali metal hydroxide solution, preferably a sodium hydroxide solution, for regenerating a single sampling column, wherein the single sampling column comprises agarose and its derivatives. Particularly preferred is agarose or cross-linked agarose, such as The fast regeneration time can guarantee the patient's treatment time, thus greatly reducing the painful time.

[0111] Matrix substrate material

[0112] According to the present invention, the compound (ligand) bound to the matrix substrate material having the property of specifically binding to the target compound (particularly CRP) is selected from the group comprising or consisting of lipids, lysophospholipids, lysophosphatidylcholine, peptides, peptides with charged amino acids, peptides containing the sequence ArgProArg, phosphorylcholine, phosphorylcholine derivatives, DNA, DNA derivatives, RNA, RNA derivatives, L-ribonucleic acid aptamers, e.g. (RNA-like molecules composed of L-ribose units), glycosides, sugars, and aptamers.

[0113] In some embodiments, the bound compound is preferably not a glycoside or a carbohydrate. According to the present invention, the bound compound is not a protein. Therefore, the compound (ligand) bound to the matrix substrate material having the property of specifically binding the target compound (especially CRP) is preferably selected from the group consisting of lipids, lysophospholipids, lysophosphatidylcholine, peptides, peptides with charged amino acids, peptides containing the sequence ArgProArg, phosphorylcholine, phosphorylcholine derivatives, DNA, DNA derivatives, RNA, RNA derivatives, L-ribonucleic acid aptamers such as (RNA-like molecules composed of L-ribose units) and aptamers.

[0114] In the use according to the invention of an alkali metal hydroxide solution, preferably sodium hydroxide solution, for regenerating an apheresis column as described herein, this is therefore preferably a CRP apheresis column.

[0115] Therefore, one embodiment of the present invention relates to the use of an alkali metal hydroxide solution (preferably a sodium hydroxide solution) to regenerate a single sampling column, wherein the single sampling column is a CRP single sampling column. In addition, another embodiment of the present invention relates to the use of an alkali metal hydroxide solution (preferably a sodium hydroxide solution) for regenerating a single sampling column for the removal of CRP by affinity chromatography.

[0116] Furthermore, another embodiment of the present invention relates to the use of an alkali metal hydroxide solution, preferably a sodium hydroxide solution, to regenerate the apheresis column, wherein the apheresis column is a CRP apheresis column and the regeneration is accomplished during the removal of CRP from the blood.

[0117] Furthermore, another embodiment of the present invention relates to the use of an alkali metal hydroxide solution, preferably a sodium hydroxide solution, for regenerating an apheresis column for the removal of CRP by affinity chromatography, wherein the regeneration is accomplished during the removal of CRP from blood.

[0118] CRP Ca 2+ Dependent ligand

[0119] For affinity chromatography to remove CRP from biological fluids (e.g. from blood or plasma), a column material containing phosphocholine and / or phosphoethanolamine or a derivative thereof may be used, thereby allowing CRP to bind to the Ca of the functionalized column material. 2+ Dependency binding.

[0120] For this purpose, phosphorylcholine, phosphorylethanolamine or its derivatives are fixed on the column material. This is usually done by an organic linker group, through which phosphorylcholine, phosphorylethanolamine or its derivatives are adsorbed or more preferably covalently bound to the column material. This produces a so-called "functionalized column material" (functionalized matrix substrate material), in which the Ca 2+ The chemical group to which the binding is dependent is exposed to the outside, so that CRP present in the biological fluid can come into contact with the chemical group.

[0121] In other words, as used herein, the term "functionalized column material" refers to a column material for affinity chromatography that has a functional chemical group (ligand). Here, the functional chemical group can be attached to the column material by adsorption or ionic interaction but preferably by a covalent bond. Of course, it is important that the functional group chemical group (ligand) is attached to the column material in such a way that the functional group is active and exposed, thereby maintaining its functionality. Therefore, the group (here: ω-phosphonoalkyl ammonium group and / or ω-alkoxy-hydroxy-phosphoryloxy group) attached to the chromatographic column material may interact or bind to the target compound (here CRP) in the sample (here: biological fluid such as blood or plasma).

[0122] Depending on whether phosphorylcholine, phosphoethanolamine or their derivatives are linked to the column material via an ammonium group or via a phosphate group through an organic linker, a distinction is made between column materials functionalized with ω-phosphonooxyalkylammonium groups (linked via ammonium groups) and column materials functionalized with ω-ammonium alkoxy-hydroxy-phosphoryloxy groups (linked via phosphate groups).

[0123] Therefore, a preferred embodiment of the present invention is to use an alkali metal hydroxide solution, preferably a sodium hydroxide solution, to regenerate a single sampling column, wherein the single sampling column comprises a matrix substrate material functionalized with at least one ω-phosphonooxyalkylammonium group and / or with at least one ω-ammonium alkoxy-hydroxy-phosphoryloxy group.

[0124] The connection to the column material, optionally via an organic linker, is shown in the following formulae (I) and (II) by a dotted line either at the nitrogen atom of the ammonium group or at the oxygen atom of the phosphate group.

[0125] As used herein, the term "ω-phosphonooxyalkylammonium group" may be used synonymously with "ω-phosphonooxyalkylammonium" and describes a compound (I) having the following general formula:

[0126]

[0127] in

[0128] n is selected from 2 and 3;

[0129] R 1 and R 2 Independently selected from: -H, -CH3, -C2H5, -C3H7, -C4H9, -C5H 11 、-C6H 13 , or R 1 and R 2 Together with the nitrogen atom to which they are attached, they can form a heterocyclic ring selected from:

[0130]

[0131] One or more hydrogen atoms may be replaced by fluorine atoms.

[0132] Thus, one embodiment of the present invention relates to the use of an alkali metal hydroxide solution, preferably a sodium hydroxide solution, to regenerate a single sampling column, wherein the single sampling column comprises a matrix substrate material functionalized with at least one ω-phosphonooxyalkylammonium group of the following general formula (I):

[0133]

[0134] in

[0135] n is selected from 2 and 3;

[0136] R 1 and R 2 Independently selected from: -H, -CH3, -C2H5, -C3H7, -C4H9, -C5H 11 、-C6H 13 , or R 1 and R 2 Together with the nitrogen atom to which they are attached, they can form a heterocyclic ring selected from:

[0137]

[0138] One or more hydrogen atoms may be replaced by fluorine atoms.

[0139] Preferably, the at least one ω-phosphonooxyalkylammonium group corresponds to a group of formula (I)

[0140]

[0141] in

[0142] n=2 or 3;

[0143] R 1 and R 2 are independently selected from: -H, -CH3, -C2H5, -C3H7, or R 1 and R 2 Together with the nitrogen atom to which they are attached, they can form a heterocyclic ring selected from:

[0144]

[0145] Particularly preferably, the at least one ω-phosphonooxyalkylammonium group corresponds to a group of the general formula (I)

[0146]

[0147] in

[0148] n = 2;

[0149] R 1 and R 2 Selected from: -H, -CH3, -C2H5, particularly preferably selected from -CH3 and -C2H5 or R 1 and R 2 Together with the nitrogen atom to which they are attached they may form a heterocyclic ring selected from:

[0150]

[0151] Preferred compounds containing an ω-phosphonooxyalkylammonium group as described above and suitable for functionalization of the corresponding column material (matrix substrate material) include, for example:

[0152] 2-[2-(2-aminoethoxy)ethyl-diethyl-ammonio]ethyl hydrogen phosphate, 2-[4-[2-(2-aminoethoxy)ethyl]morpholin-4-ium-4-yl]ethyl hydrogen phosphate, 2-[1-[2-(2-aminoethoxy)ethyl]piperidin-1-ium-1-yl]ethyl hydrogen phosphate, 2-[2-(2-aminoethoxy)ethyl-dimethyl-ammonio]ethyl hydrogen phosphate, 2-[3-aminopropyl-(dimethyl)ammonio]ethyl hydrogen phosphate, 2-[dimethyl(4-sulfanylbutyl)ammonio]ethyl hydrogen phosphate, 2-[4-azidobutyl(dimethyl)ammonio]ethyl hydrogen phosphate, 2-[dimethyl(pentan-4 -alkynyl)ammonio]ethyl hydrogen phosphate, 2-[3-(6-aminohexanoyl-amino)propyl-diethyl-ammonio]ethyl hydrogen phosphate, 2-[1-[2-[2-(6-aminohexanoyl-amino))ethoxy]ethyl]piperidin-1-ium-1-yl]ethyl hydrogen phosphate, 2-[4-[2-[2-[3-(6-aminohexanoylamino)propionylamino]ethoxy]ethyl]morpholin-4-ium-4-yl]ethyl hydrogen phosphate, 2-[1-[2-[2-[6-(6-aminohexanoylamino)hexanoylamino]ethoxy]ethyl]pyrrolidin-1-ium-1-yl]ethyl hydrogen phosphate, 2-[2-allyloxyethyl (dimethyl)ammonium 2-[2-allyloxyethyl (diethyl) ammonium] ethyl hydrogen phosphate, 2-[4-(2-allyloxyethyl) morpholinium-4-yl] ethyl hydrogen phosphate, 2-[1-(2-allyloxyethyl) piperidin-1-yl] ethyl hydrogen phosphate, 2-[2-[2-(6-aminohexanoylamino) ethoxy] ethyl dimethyl-ammonium] ethyl hydrogen phosphate, 2-[2-[2-[3-(6-aminohexanoylamino) propionylamino] ethoxy] ethyl-dimethyl-ammonium] ethyl hydrogen phosphate, 2-[3-azidopropyl (dimethyl) ammonium] ethyl hydrogen phosphate, 2-[dimethyl-[2-[2 -(prop-2-ynyloxycarbonylamino)ethoxy]ethyl]ammonio]ethyl hydrogen phosphate, 2-[2-[2-(allyloxycarbonylamino)ethoxy]ethyldimethyl-ammonio]ethyl hydrogen phosphate, 2-[2-[2-[6-(allyloxycarbonylamino)hexanoylamino]ethoxy]ethyldimethyl-ammonio]ethyl hydrogen phosphate, 2-[2-(6-aminohexanoylamino)ethyl-dimethyl-ammonio]ethyl hydrogen phosphate, 2-[dimethyl-[3-[6-(prop-2-ynyloxycarbonylamino)hexanoylamino]propyl]ammonio]ethyl hydrogen phosphate and 2-[3-(6-aminohexanoylamino)propyl-dimethyl-ammonio]ethyl hydrogen phosphate.

[0153] As used herein, the term "ω-ammonium alkoxy-hydroxy-phosphoryloxy group" may be used synonymously with "ω-ammonium alkoxy-hydroxy-phosphoryloxy" and describes the following compounds of formula (II):

[0154]

[0155] in

[0156] n is selected from 2 and 3;

[0157] R 1 , R 2 and R 3 are independently selected from: -H, -CH3, -C2H5, -C3H7, -C4H9, -C5H 11 , –C6H 13 ,

[0158] or R 1 and R 2 Together with the nitrogen atom to which they are attached, they can form a heterocyclic ring selected from:

[0159]

[0160] and

[0161] R 3 Selected from: -H, -CH3, -C2H5, -C3H7, -C4H9, -C5H 11 、-C6H 13 , and preferably -H;

[0162] One or more hydrogen atoms may be replaced by fluorine atoms.

[0163] Preferred "ω-ammonium alkoxy-hydroxy-phosphoryloxy" compounds include compounds of the general formula (II)

[0164]

[0165] in

[0166] n is selected from 2 and 3;

[0167] R 1 , R 2 and R 3 are independently selected from: -H, -CH3, -C2H5, -C3H7,

[0168] or R 1 and R 2 Together with the nitrogen atom to which they are attached, they can form a heterocyclic ring selected from:

[0169]

[0170] And R 3 =H.

[0171] In the context of the present invention, it is particularly preferred that the ω-ammonium alkoxy-hydroxy-phosphoryloxy group is an ω-trialkylammonium alkoxy-hydroxy-phosphoryloxy group.

[0172] Thus, particularly preferred ω-ammonium alkoxy-hydroxy-phosphoryloxy groups include compounds of the general formula (II)

[0173]

[0174] in

[0175] n = 2;

[0176] R 1 , R 2 and R 3 Selected from: -H, -CH3, -C2H5, particularly preferably selected from -CH3 and -C2H5.

[0177] It is also particularly preferred if the ω-ammonium alkoxy-hydroxy-phosphoryloxy group is an ω-trimethylammonium ethoxy-hydroxy-phosphoryloxy group or an ω-trimethylammonium propoxy-hydroxy-phosphoryloxy group.

[0178] Preferred compounds which contain an ω-ammonium alkoxy-hydroxy-phosphoryloxy group as described above and are suitable for functionalization of the corresponding column materials include, for example: p-aminophenylphosphorylcholine (APPC), 4[[hydroxy[2-(trimethylammonium)ethoxy]phosphonyl]oxy]benzenediazonium (p-diazoniophenylphosphorylcholine) or p-nitrophenyl 6-(O-phosphorylcholine) hydroxyhexanoate.

[0179] Therefore, a preferred embodiment relates to the use of an alkali metal hydroxide solution, preferably a sodium hydroxide solution, for regenerating a single sampling column, wherein the single sampling column comprises a matrix material functionalized with at least one ω-ammonium alkoxy-hydroxy-phosphoryloxy group corresponding to the group of the following general formula (II):

[0180]

[0181] in

[0182] n is selected from 2 and 3;

[0183] R 1 , R 2 and R 3 Independently selected from: -H, -CH3, -C2H5, -C3H7, -C4H9, -C5H 11 、-C6H 13 ,

[0184] or R 1 and R 2 Together with the nitrogen atom to which they are attached, they can form a heterocyclic ring selected from:

[0185]

[0186] and

[0187] R 3 Selected from: -H, -CH3, -C2H5, -C3H7, -C4H9, -C5H 11 、-C6H 13 , and preferably -H;

[0188] One or more hydrogen atoms may be replaced by fluorine atoms.

[0189] Another aspect of the present invention relates to a method for regenerating an apheresis column for the removal of CRP by affinity chromatography, comprising the following steps:

[0190] (I) An alkali metal hydroxide solution (preferably a sodium hydroxide solution) is introduced into the single sampling column for regeneration of the single sampling column.

[0191] The alkali metal hydroxide solution (preferably sodium hydroxide solution) can be removed from the apheresis column by a neutralizing solution to prepare the apheresis column for apheresis treatment. Therefore, the method according to the present invention can include a step (II) of introducing a neutralizing solution.

[0192] Therefore, one embodiment of the present invention relates to a method for regenerating an apheresis column for removing CRP by affinity chromatography, comprising the following steps:

[0193] (I) introducing an alkali metal hydroxide solution (preferably a sodium hydroxide solution) into the single sampling column for regeneration of the single sampling column; and

[0194] (II) Introducing a neutralizing solution.

[0195] Therefore, one embodiment of the present invention relates to a method for regenerating an apheresis column for removing CRP by affinity chromatography, comprising the following steps:

[0196] (I) introducing an alkali metal hydroxide solution (preferably a sodium hydroxide solution) into the single sampling column for regeneration of the single sampling column;

[0197] (II') after step (I), the introduction of the alkali metal hydroxide solution (preferably sodium hydroxide solution) is stopped and

[0198] (II) Introducing a neutralizing solution.

[0199] Regeneration of the apheresis column can be performed during the apheresis treatment. In this case, plasma loss can be minimized by removing plasma from parts of the apheresis device, in particular from the apheresis column, before the introduction of the alkali metal hydroxide solution, preferably sodium hydroxide solution, for which purpose a flushing solution can be used. Therefore, the method according to the invention may comprise the step (I') of flushing the apheresis column, preferably containing plasma.

[0200] Therefore, one embodiment of the present invention relates to a method for regenerating an apheresis column for removing CRP by affinity chromatography, comprising the following steps:

[0201] (I') introducing a flushing solution into the apheresis column, which preferably contains plasma;

[0202] (I) introducing an alkali metal hydroxide solution (preferably a sodium hydroxide solution) into the single sampling column for regeneration of the single sampling column;

[0203] (II') stopping the introduction of the alkali metal hydroxide solution (preferably sodium hydroxide solution) after step (I); and

[0204] (II) Introducing a neutralizing solution.

[0205] Flushing solution

[0206] The flushing solution can, but need not necessarily, be used for regeneration of the apheresis column, but, in addition to the above functions, has the task of replacing plasma from the apheresis column or for neutralizing the apheresis column matrix. As a flushing solution, a sodium chloride solution, in particular a physiological sodium chloride solution or a PBS solution (phosphate buffered saline) can be used. Preferably, a NaCl solution, particularly preferably a physiological NaCl solution, is used as a flushing solution.

[0207] As used herein, the term "NaCl solution" (sodium chloride solution) includes an aqueous solution containing sodium chloride (i.e., NaCl, also referred to as table salt) as a main component. As used herein, "main component" refers to a NaCl solution in which the molar concentration of sodium chloride is greater than the corresponding molar concentrations of all other compounds in the NaCl solution, but does not include water. Preferably, the NaCl solution comprises 0.1 to 5 wt % sodium chloride, particularly preferably 0.9 wt %. Preferably, the flushing solution is such a NaCl solution. Physiological NaCl solution (PBS solution) is understood to be a sodium chloride solution comprising water and 0.9 wt % sodium chloride (NaCl).

[0208] Neutralizing liquid

[0209] As used herein, the term "neutralizing solution" refers to an aqueous solution used to adjust the pH range to 6.5 to 7.6, preferably 7.30 to 7.50, more preferably 7.35 to 7.45.

[0210] In principle, all aqueous solutions can be considered as neutralizing solutions that are allowed to be used in the medical field. Preferably, the pH of the aqueous solution is ≤7, that is, the aqueous solution can have a neutral pH or a pH <7. Preferably, the neutralizing solution is selected from the group comprising or consisting of the following: PBS solution or NaCl solution or citrate solution. Particularly preferably, the neutralizing solution is a citrate solution. Therefore, another aspect of the present invention relates to the use of a citrate solution for neutralizing a single extraction column with a column material, wherein the column material is present in an alkaline medium. Therefore, the method according to the present invention can include step (II) introducing a neutralizing solution, wherein the neutralizing solution is PBS, NaCl or a citrate solution, preferably a citrate solution.

[0211] In addition, one embodiment of the present invention relates to a method for regenerating a single sampling column for removing CRP by affinity chromatography, comprising the following steps:

[0212] (I) introducing an alkali metal hydroxide solution (preferably a sodium hydroxide solution) into the single sampling column for regeneration of the single sampling column;

[0213] (II') stopping the introduction of the alkali metal hydroxide solution (preferably sodium hydroxide solution) after step (I); and

[0214] (II) Introducing PBS solution, NaCl solution or citrate solution.

[0215] As used herein, the term "citrate solution" includes aqueous solutions containing at least one citrate compound.

[0216] As used herein, the term "citrate" relates to a citrate anion, i.e. a salt of citric acid. Preferably, the citrate solution contains at least one citrate compound selected from the group comprising or consisting of citric acid, sodium dihydrogen citrate, disodium hydrogen citrate, trisodium citrate, trisodium citrate dihydrate, potassium dihydrogen citrate, dipotassium hydrogen citrate, tricalcium citrate, lithium dihydrogen citrate, dilithium hydrogen citrate, trilithium citrate, ammonium dihydrogen citrate, diammonium hydrogen citrate, triammonium citrate, tricalcium dicitrate (calcium citrate), trimagnesium dicitrate (magnesium citrate) and / or a partial ester of citric acid.

[0217] The citrate solution consisting of citric acid, trisodium citrate, D-glucose and water is also referred to as "acid citrate dextrose solution (ACD solution)". A preferred variant of the citrate solution used according to the invention relates to an ACD solution containing between 22.9 mM and 38.0 mM citric acid, between 44.9 mM and 74.8 mM trisodium citrate, between 74.2 mM and 123.6 mM D-glucose and water. A particularly preferred variant of the citrate solution used according to the invention relates to an ACD solution containing 38 mM citric acid, 74.8 mM trisodium citrate, 123.6 mM D-glucose and water. This citrate solution is also referred to as "ACD-A solution".

[0218] The citrate solution composed of citric acid, trisodium citrate, sodium hydrogen phosphate, D-glucose and water is also called "citrate-phosphate-dextrose solution (CPD)". The citrate solution composed of citric acid, trisodium citrate, sodium hydrogen phosphate, D-glucose, adenine and water is also called "citrate-phosphate-dextrose solution with adenine (CPDA)".

[0219] Preferably, the citrate solution has a concentration of 2-40%, preferably 4%, and has a pH in the range of 6.4-7.5. Preferably, a flow rate of 80 ml / min is used. The advantages of using a citrate solution compared to a PBS solution are reduced neutralization time and reduced flushing volume required.

[0220] Device

[0221] Another aspect of the invention relates to an apheresis device (1) for extracorporeal removal of CRP from a patient's blood, wherein the apheresis device (1) is configured to tolerate alkali metal hydroxide solutions, such as sodium hydroxide solutions. Of course, only those parts of the apheresis device that come into contact with alkali metal hydroxide solutions, i.e. those parts where alkali metal hydroxide solutions are stored or through which alkali metal hydroxide solutions flow, must be tolerant to the alkali metal hydroxide solutions used. For example, a cell separator and a bypass line do not come into contact with alkali metal hydroxide solutions and therefore do not have to be tolerant to the alkali metal hydroxide solutions used.

[0222] As used herein, the term "tolerance" means that the product properties do not change, for example in terms of biocompatibility and performance.

[0223] Chemical resistance generally refers to the resistance of a material to exposure to chemicals. A material is chemically resistant if its mechanical, physical and chemical properties remain unchanged or are only very slowly corroded even after a long period of contact with the chemical to be tested. A material has a particular chemical resistance if its characteristic properties remain unchanged for a limited period of time acceptable for the purpose of use or within specific restrictions of the conditions of use. On the other hand, a chemically unstable material is a material that loses its characteristic properties in a very short time or faster than the purpose of use allows. Therefore, as used herein, resistance also preferably means that the characteristic properties of the material of the part of the single sampling device that is in contact with the alkali metal hydroxide solution, preferably the sodium hydroxide solution, remain unchanged during a contact time of at least 20 hours. Typically, the acceptable time period for the purpose of use is between 4 hours and 8 hours.

[0224] Therefore, the parts in contact with the alkali metal hydroxide solution (preferably sodium hydroxide solution) must be made of materials that tolerate alkali metal hydroxide (preferably sodium hydroxide). Suitable materials include, but are not limited to, stainless steel, polypropylene (PP), polyvinyl chloride (PVC), polyethylene (PE), polyethylene terephthalate (PET) and polycarbonate (PC). Preferred stainless steel, polypropylene (PP), polyvinyl chloride (PVC), polyethylene (PE) and polyethylene terephthalate (PET). Here, it is considered that the materials that tolerate alkali metal hydroxide (preferably sodium hydroxide) include, but are not limited to, stainless steel, polypropylene (PP), polyvinyl chloride (PVC), polyethylene (PE), polyethylene terephthalate (PET) and polycarbonate (PC). The preferred materials that tolerate alkali metal hydroxide (preferably sodium hydroxide) are selected from stainless steel, polypropylene (PP), polyvinyl chloride (PVC), polyethylene (PE), polyethylene terephthalate (PET).

[0225] In other words, another aspect of the present invention relates to an apheresis device (1) for the extracorporeal removal of CRP from the blood of a patient, wherein the apheresis device is connectable to the blood circulation of the patient, and wherein the apheresis device (1) is designed to be resistant to alkali metal hydroxide solutions, preferably sodium hydroxide solutions. That is, the mechanical, physical and chemical properties of the parts of the apheresis device that are in contact with the alkali metal hydroxide solution do not change during the operating time of the apheresis device for apheresis treatment.

[0226] For the regeneration of the single sampling column, an alkali metal hydroxide solution (preferably a sodium hydroxide solution) can be used. The alkali metal hydroxide solution (preferably a sodium hydroxide solution) is known for its reactivity, which, however, depends largely on the concentration of the alkali metal hydroxide in the alkali metal hydroxide solution, preferably the sodium hydroxide in the sodium hydroxide solution. In addition, other compounds in the alkali metal hydroxide solution (preferably a sodium hydroxide solution) influence the reactivity of the alkali metal hydroxide, preferably sodium hydroxide (or sodium hydroxide in dissociated form).

[0227] Thus, alkali metal hydroxide solutions, preferably sodium hydroxide solutions, attack the material. For example, a strong base such as alkali metal hydroxide solutions, preferably sodium hydroxide, reacts with the glass, which leads to dissolution of glass components. There are also materials made of organic polymers that are not resistant to the presence of alkali metal hydroxide solutions, preferably sodium hydroxide, at room temperature or higher.

[0228] An embodiment according to the present invention relates to an apheresis device (1) for extracorporeal removal of CRP from a patient's blood, wherein the apheresis device can be connected to the patient's blood circulation. Blood is pumped to a cell separator (7) through a part of the extracorporeal circulation system (2) of the apheresis device (1) according to the present invention for separation of the blood into plasma and cellular components. Through a first outlet of the cell separator (7), the separated plasma is guided to an apheresis column (4) through a plasma line (8A) for affinity chromatography removal of CRP from the plasma. After the CRP is removed from the patient's plasma, this treated plasma is combined with the cellular components in the blood through a plasma line (8B). In addition, the apheresis device (1) according to the present invention comprises a bypass line (12) that bypasses the apheresis column (4) and leads from the plasma line (8A) to the plasma line (8B). In addition, the apheresis device (1) according to the present invention comprises at least one regeneration line (14), which is connected to the plasma line (8A) or directly to the apheresis column (4) after the bypass line (12) in the flow direction. The apheresis device (1) is designed to be resistant to alkali hydroxide solutions, preferably sodium hydroxide solutions.

[0229] That is, the portion of the apheresis device that comes into contact with the alkali metal hydroxide solution is resistant to the alkali metal hydroxide solution used.

[0230] One embodiment of the present invention relates to an apheresis device (1) for extracorporeal removal of CRP from blood, comprising:

[0231] Extracorporeal circulation system for blood (2),

[0232] Device (3) for generating and regulating blood flow in an extracorporeal circulation system (2),

[0233] a cell separator (7) for separating blood into plasma and cellular components,

[0234] at least one apheresis column (4) for affinity chromatographic removal of CRP from blood,

[0235] The extracorporeal circulation system (2) includes an arterial line (5) to a cell separator (7), a plasma line (8A) from the cell separator (7) to a single collection column (4), a plasma line (8B) for CRP-depleted plasma from the single collection column (4) to a node (P1), a cell line (9) for separated cellular components from the cell separator (7) to the node (P1), and a venous line (6) from the node (P1).

[0236] at least one connecting line (11) for connecting at least one liquid container (F) to the arterial line (5) or to the cell separator (7),

[0237] It is characterized in that

[0238] a bypass line (12) branching from the plasma line (8A) and connected to the plasma line (8B),

[0239] a waste liquid pipeline (13) which is directly separated from the single collection column (4) or separated from the plasma pipeline (8B) before the connection point of the bypass pipeline (12) in the flow direction, and

[0240] At least one regeneration line (14) leading to the plasma line (8A) or directly connected to the single sampling column (4) at the branch of the bypass line (12) or after it in the flow direction, wherein the single sampling device (1) is designed to be resistant to alkali metal hydroxide solutions (preferably sodium hydroxide solutions).

[0241] Preferably, the apheresis device (1) further comprises a central processing unit (10) for controlling the apheresis device (1).

[0242] One embodiment of the present invention relates to an apheresis device (1) for extracorporeal removal of CRP from blood, comprising:

[0243] Extracorporeal circulation system for blood (2),

[0244] Device (3) for generating and regulating blood flow in an extracorporeal circulation system (2),

[0245] a cell separator (7) for separating blood into plasma and cellular components,

[0246] at least one apheresis column (4) for affinity chromatographic removal of CRP from blood,

[0247] The extracorporeal circulation system (2) includes an arterial line (5) to a cell separator (7), a plasma line (8A) from the cell separator (7) to a single collection column (4), a plasma line (8B) for CRP-depleted plasma from the single collection column (4) to a node (P1), a cell line (9) for separated cellular components from the cell separator (7) to the node (P1), and a venous line (6) from the node (P1).

[0248] at least one connecting line (11) for connecting at least one liquid container (F1) to the arterial line (5) or to the cell separator (7),

[0249] It is characterized in that

[0250] a bypass line (12) branching from the plasma line (8A) and connected to the plasma line (8B),

[0251] a waste liquid pipeline (13) which is directly separated from the single collection column (4) or separated from the plasma pipeline (8B) before the connection point of the bypass pipeline (12) in the flow direction,

[0252] At least one regeneration line (14) branches out from at least one liquid container (F1) or at least one connecting line (11) and leads to the plasma line (8A) or directly connects to the single sampling column (4) at or after the branch of the bypass line (12) in the flow direction, and

[0253] At least one second regeneration line (14) branches out from at least one liquid container (F2), wherein the second regeneration line (14) is not connected to the arterial line (5) or the cell separator (7), and is led to the plasma line (8A) or directly connected to the single collection column (4) at or after the branch of the bypass line (12) in the flow direction,

[0254] The single sampling device (1) is designed to be resistant to alkali hydroxide solution (preferably sodium hydroxide solution).

[0255] Preferably, the apheresis device (1) further comprises a central processing unit for controlling the apheresis device (1).

[0256] Preferably, the single collection device (1) therefore comprises at least two regeneration lines (14', 14") which are independently led in the flow direction at or after the branch of the bypass line (12) to the plasma line (8A) or directly connected to the single collection column (4'), wherein at least one of the regeneration lines (14', 14") branches off from at least one liquid container (F1) or at least one connecting line (11).

[0257] One embodiment of the present invention relates to an apheresis device (1) for extracorporeal removal of CRP from blood, comprising:

[0258] Extracorporeal circulation system for blood (2),

[0259] Device (3) for generating and regulating blood flow in an extracorporeal circulation system (2),

[0260] a cell separator (7) for separating blood into plasma and cellular components,

[0261] at least one apheresis column (4) for affinity chromatographic removal of CRP from blood,

[0262] The extracorporeal circulation system (2) includes an arterial line (5) to a cell separator (7), a plasma line (8A) from the cell separator (7) to a single collection column (4), a plasma line (8B) for CRP-depleted plasma from the single collection column (4) to a node (P1), a cell line (9) for separated cellular components from the cell separator (7) to the node (P1), and a venous line (6) from the node (P1).

[0263] at least one connecting line (11) for connecting at least one liquid container (F1) to the arterial line (5) or to the cell separator (7),

[0264] It is characterized in that

[0265] a bypass line (12) branching from the plasma line (8A) and connected to the plasma line (8B),

[0266] a waste liquid pipeline (13) which is directly separated from the single collection column (4) or separated from the plasma pipeline (8B) before the connection point of the bypass pipeline (12) in the flow direction, and

[0267] At least one regeneration line (14) leads from at least one liquid container (F1) or at least one connecting line (11) and leads in the flow direction at or after a branch of a bypass line (12) to a plasma line (8A) or directly to a single sampling column (4), wherein at least one regeneration line (14) has at least one additional connection for a liquid container (F2), wherein the single sampling device (1) is designed to be resistant to alkali metal hydroxide solutions (preferably sodium hydroxide solutions).

[0268] Preferably, the apheresis device (1) further comprises a central processing unit for controlling the apheresis device (1).

[0269] As mentioned above, the term "resistant" is used to mean that only those parts of the apheresis device that come into contact with the alkali metal hydroxide solution must be resistant or resistant to the alkali metal hydroxide solution used. The rest of the apheresis device may, but need not, be resistant to the alkali metal hydroxide solution used.

[0270] Tolerance means that the biocompatibility according to the DIN EN ISO 10993-1 to -12 standard series does not change. The DIN EN ISO 10993-1 to -12 standard series includes ISO 10993-1 Evaluation and testing within the risk management approach, ISO10993-2 Animal protection requirements, ISO 10993-3 Genotoxicity, carcinogenicity and reproductive toxicity tests, ISO 10993-4 Selection of tests for interaction with blood, ISO 10993-5 In vitro cytotoxicity tests, ISO 10993-6 Tests for local effects after implantation, ISO 10993-7 Ethylene oxide sterilization residues, ISO 10993-8 Selection and suitability of reference materials for biological tests, ISO 10993-9 Framework for identification and quantification of possible degradation products, ISO 10993-10 Irritation and skin sensitization tests, ISO10993-11 Systemic toxicity tests, ISO 10993-12 Sample preparation and reference materials. This can be done by testing extracts. For the qualitative analysis of the extracts obtained, the standard recommends the use of gas chromatography (GC) or (high performance) liquid chromatography (LC or HPLC) in combination with mass spectrometry (MS). For further analysis, the identification of typically extracted compounds with the aid of ion pair chromatography (IPC) or the identification of extractable metal ions with the aid of inductively coupled plasma (ICP) is recommended. Cytotoxicity tests, hemocompatibility tests and extractable component tests are selected from a number of possible test methods in accordance with DIN EN ISO 10993. Cytocompatibility tests check whether the product has toxic / harmful effects on cells. The test is carried out under direct and / or indirect contact and reliably identifies toxic substances. In vitro hemocompatibility tests detect undesirable material properties at an early stage, before the product causes harm to the patient. The tests are carried out with human blood in simple static or complex dynamic systems. Biomaterials and medical devices trigger nonspecific foreign body reactions in vivo. Local tissue reaction tests determine parameters for assessing the suitability and value of alternative materials.

[0271] Preferably, therefore, the apheresis device according to the invention is configured to be resistant to alkali metal hydroxide solutions, preferably sodium hydroxide solutions, as described herein, wherein resistance to alkali metal hydroxide solutions, preferably sodium hydroxide solutions, is present if the biocompatibility according to the DIN EN ISO 10993-1 to -12 standard series is not altered.

[0272] Therefore, preferably, as described herein, the apheresis device according to the present invention is configured to be resistant to alkali metal hydroxide solutions (preferably sodium hydroxide solutions), wherein the resistance to alkali metal hydroxide solutions (preferably sodium hydroxide solutions) exists if the components in contact with the alkali metal hydroxide solutions (preferably sodium hydroxide solutions) are made of a material resistant to alkali metal hydroxides (preferably sodium hydroxide), wherein the material resistant to alkali metal hydroxides (preferably sodium hydroxide) is selected from the group comprising or consisting of stainless steel, polypropylene (PP), polyvinyl chloride (PVC), polyethylene (PE), polyethylene terephthalate (PET) and / or polycarbonate (PC), preferably stainless steel, polypropylene (PP), polyvinyl chloride (PVC), polyethylene (PE) and / or polyethylene terephthalate (PET).

[0273] Preferably, the plasma line is made of stainless steel or a polymer selected from the group consisting of polypropylene (PP), polyvinyl chloride (PVC), polyethylene (PE), polyethylene terephthalate (PET) and polycarbonate (PC). Particularly preferably, the plasma line is made of stainless steel or a polymer selected from the group consisting of polypropylene (PP), polyvinyl chloride (PVC), polyethylene (PE) and polyethylene terephthalate (PET).

[0274] Preferably, the liquid container for containing the regeneration solution is made of stainless steel or a polymer selected from the group consisting of (PP), polyvinyl chloride (PVC), polyethylene (PE), polyethylene terephthalate (PET) and polycarbonate (PC). Particularly preferably, the liquid container for containing the regeneration solution is made of stainless steel or a polymer selected from the group consisting of (PP), polyvinyl chloride (PVC), polyethylene (PE) and polyethylene terephthalate (PET).

[0275] Further preferably, the housing of the single sampling column is made of stainless steel or a polymer selected from the group consisting of: (PP), polyvinyl chloride (PVC), polyethylene (PE), polyethylene terephthalate (PET) and polycarbonate (PC). Particularly preferably, the housing of the single sampling column is made of stainless steel or a polymer selected from the group consisting of: stainless steel, polypropylene (PP), polyvinyl chloride (PVC), polyethylene (PE) and polyethylene terephthalate (PET).

[0276] As described above, the apheresis device (1) for extracorporeal removal of CRP from blood according to the present invention can be connected to the patient's blood circulation. From a vascular access point (usually a venous access point) in the patient, blood is pumped to the cell separator (7) through a portion of the extracorporeal circulation system (2) of the present invention. The portion of the extracorporeal circulation system (2) that draws blood from the patient's body and flows to the cell separator (7) draws blood from the patient's body away from the patient's heart and is therefore referred to as an "arterial line" (5) in reference to the nomenclature of the blood vessels of the human body.

[0277] The patient's blood is introduced into the cell separator (7) via its inlet and separated therein into plasma (sometimes also referred to as "plasma") and cellular components of the blood. It must be taken into account that the separation of plasma and cellular components is not complete, but that only preferably 10-90% of the total plasma is separated from the cellular components. Through the first outlet of the cell separator (7), the separated plasma is guided via a plasma line (8A) to a single collection column (4) for affinity chromatography to remove CRP from the blood (or plasma). After the removal or reduction of CRP in the patient's plasma, the now treated plasma (also referred to as "depleted plasma") is guided via a plasma line (8B) to a node (P1). Through the second outlet of the cell separator (7) and a connecting line (the so-called cell line (9)), the cellular components in the blood pass by the single collection column (4) and are guided to the node (P1). There, the cellular components merge with the depleted plasma. After the cellular components have been combined with the depleted plasma, the now treated blood is returned to the patient via another part of the extracorporeal circulation system (2) of the invention. The part of the extracorporeal circulation system (2) that conducts the treated blood from point (P1) of the extracorporeal circulation system (2) back to the patient conducts the blood to the patient and thus also to the patient's heart and is therefore referred to as a "venous line" (6) in reference to the nomenclature of the blood vessels of the human body.

[0278] In an alternative embodiment of the invention it is possible that the cellular components can also be returned directly to the patient after separation from the plasma via a second outlet on the cell separator and the subsequent line and that only the processed plasma is conveyed to the patient via the venous line.

[0279] In order to be able to prevent blood clotting in the extracorporeal circulation system or to be able to flush or pre-flush the extracorporeal circulation system (for example with physiological saline solution), the apheresis device according to the invention comprises at least one line (so-called Connecting pipeline(11)), which is capable of connecting at least one liquid container (F) and thus being capable of supplying the liquid contained in the at least one liquid container (e.g., an anticoagulant or a physiological saline solution) to the extracorporeal circulation system. In this case, it also means that the connecting line (11) for connecting the at least one liquid container (F) is fluidically connected to the extracorporeal circulation system, that is, the liquid from the liquid container can be introduced into the extracorporeal circulation system via the connecting line (11). In a preferred embodiment of the present invention, at least one connecting line (11) is connected to the extracorporeal circulation system (2) before the cell separator (7), that is, enters the arterial line (5), or directly enters the cell separator (7).

[0280] It is obvious to a person skilled in the art that the liquid container (F) itself does not have to be part of the apheresis device according to the invention, since these are usually disposable items, e.g. in the form of a common infusion bag, which is connected to the connecting line by the operator (e.g. the attending physician or nurse) according to the specific application.

[0281] According to the invention, it is possible that there is a single connecting line (11) for connecting a liquid container. However, it is also conceivable that there is a single connecting line (11) to which two or three or preferably more liquid containers can be connected. Similarly, it is possible that an embodiment of the apheresis device according to the invention has two, preferably three or preferably more connecting lines (11', 11", 11'", etc.), each for connecting at least one liquid container, wherein preferably, these two, preferably three or preferably more connecting lines can be connected independently of each other to the arterial line (5) or directly to the cell separator (7). “Independent of each other” in this context means, for example, in an embodiment of the single collection device according to the present invention having two connecting lines (11', 11"), one connecting line (11') can be connected to the arterial line (5) and the other connecting line (11") can be directly connected to the cell separator (7), and both connecting lines (11', 11") can enter the arterial line (5) or both connecting lines (11', 11") can be directly connected to the cell separator (7).

[0282] According to an embodiment of the present invention, it is particularly preferred that the single collection device (1) according to the present invention has two connecting lines (11', 11"), each of which is used to connect at least one liquid container, wherein the connecting lines (11', 11") are independently connected to the arterial line (5) or directly connected to the cell separator (7). Therefore, the two connecting lines (11', 11") are both connected to the arterial line (5), or the two connecting lines (11', 11") are directly connected to the cell separator (7), or particularly preferably, one connecting line (11') is connected to the arterial line (5) and the other connecting line (11") directly enters the cell separator (7). Therefore, the two connecting lines (11', 11") can be connected to different liquid containers. It is particularly preferred that one of the two connecting lines (e.g. 11') is connected to a liquid container containing a physiological salt solution (e.g. NaCl solution), and the second of the two connecting lines (e.g. 11") is connected to a liquid container containing, for example, a citrate solution.

[0283] Therefore, it is particularly preferred that the apheresis device (1) has a connecting line (11') for connecting to the liquid container (F1) and a connecting line (11") for connecting to the liquid container (F2), and the line (11') is connected to the arterial line (5) or the cell separator (7), the connecting line (11") is connected to the arterial line (5) or the cell separator (7) or the connecting line (11') and thus ultimately also to the arterial line (5) or the cell separator (7).

[0284] A significant advantage of the apheresis device according to the invention is that the apheresis column, which is naturally limited in purification capacity, can be regenerated during continuous operation, i.e. without having to stop the blood discharge and the blood introduction or the cell separator. For this purpose, there is a diversion which allows the plasma flow to bypass the apheresis column (4). Bypass line (12, also called "shunt"). The bypass line (12) enables the single collection column (4) to be temporarily disconnected from the plasma flow, thereby achieving the regeneration of the single collection column (4) without interrupting the flow of blood or plasma in the device according to the present invention. The bypass line branches off from the plasma line (8A), wherein the point where the bypass line branches off from the plasma line (8A) is called point (P2), and is preferably connected to the plasma line (8B), wherein the point where the bypass line (12) connects to the plasma line (8B) is called point (P6). In an equally possible embodiment, the bypass line (12) is not connected to the plasma line (8B), but to the cell line (9), wherein the point where the bypass line (12) connects to the cell line (9) is called point (P3).

[0285] The regeneration solution required for single sampling column regeneration is Regeneration line (14)Sent into the extracorporeal circulation system (2), wherein the regeneration line (14) is either directly connected to the single collection column (4) or (along the flow direction) connected to the plasma line (8A) before the single collection column (4) but (along the flow direction) at the branch of the bypass line or after it, that is, after point (P2).

[0286] In order to remove the regeneration solution from the system after passing through the apheresis column (4) (and not deliver it to the patient), there is a branch from the plasma line (8B) Waste line (13), wherein the point in the plasma line (8B) from which the waste liquid line (13) branches is referred to as point (P4). In an embodiment in which the bypass line (12) is connected to the cell line (9), point (P4) is preferably located in the region from the single collection column (4) to point (P1). In an embodiment in which the bypass line (12) is connected to the plasma line (8B), point (P4) is preferably located in the region from the single collection column (4) to point (P6). The waste liquid line (13) can of course be connected to, for example, a collecting container. Alkali metal hydroxide solution (preferably sodium hydroxide solution) is particularly preferred as the regeneration solution according to the present invention.

[0287] In addition to the regeneration solution, a flushing solution can also be used. The flushing solution can be, but is not necessarily, used to regenerate the single sampling column (4), but has the main task of replacing the plasma in the plasma line (8A) in the area from point P2 to the single sampling column (4), in the single sampling column (4), and in the plasma line (8B) from the single sampling column (4) to point P4 before using the regeneration solution, and after circulating through the single sampling column (4), it is discarded through the waste liquid line (13). Preferably, a physiological NaCl solution or a PBS solution is used as a flushing solution. Even more preferably, if an alkali metal hydroxide solution (preferably a sodium hydroxide solution) is used as the regeneration solution, a physiological NaCl solution is used as the flushing solution.

[0288] Preferably, the single collection device (1) includes at least one regeneration line (14), which leads to the plasma line (8A) or directly to the single collection column (4) at or after the branch of the bypass line (12) in the flow direction. Preferably, the single collection device (1) includes at least one regeneration line (14), which is connected to the extracorporeal circulation system (2) in the area from the branch of the bypass line (12) at the plasma line (8A) to the single collection column (4). Preferably, the single collection device (1) includes at least one regeneration line (14), which is connected to the extracorporeal circulation system (2) in the area from point (P2) to the single collection column (4).

[0289] According to an embodiment of the present invention, preferably, the apheresis device (1) according to the present invention has at least two connecting lines (11), which respectively connect at least one liquid container to the arterial line (5) or the cell separator (7).

[0290] In addition, an embodiment of the single collection device (1) is preferred, wherein the single collection device (1) has at least two connecting pipelines (11), respectively connecting at least one liquid container (F) to the arterial line (5) or the cell separator (7), and wherein each liquid container (F) has a regeneration line (14), which leads out from each liquid container (F) or their connecting pipelines (11) and each leads to the plasma line (8A) or directly introduced into the single collection column (4).

[0291] Furthermore, it is possible that at least two connecting lines (11) merge before their connection, ie merge into one line. It is also possible that the regeneration lines (14) merge before their connection, ie merge into one line.

[0292] If it is described in the present application that a device feature is in the region from the first position in the device to the second position in the device, or is connected to the region or branches out from the region, it should be understood that the first position and the second position and the portion located therebetween are all surrounded by the region. This can be illustrated by the following example: the statement that the "regeneration line (14) is connected to the extracorporeal circulation system (2) in the region from point (P2) to the single sampling column (4)" refers to the region where the regeneration line (14) is connected to the extracorporeal circulation system (2), which includes not only the portion between point (P2) and the single sampling column (4), but also point (P2) itself and the single sampling column (4). This means that the regeneration line (14) can be connected to point (P2), or to the single sampling column (4), or even to the portion of the plasma line (8A) located between point (P2) and the single sampling column (4).

[0293] Point (P1) is the node where the plasma line (8B) and the venous line (6) merge in the extracorporeal circulation system (2). Point (P2) is the node in the extracorporeal circulation system (2) where the bypass line (12) branches off from the plasma line (8A). Point (P3) is the node in the extracorporeal circulation system (2) where the bypass line (12) connects to the cell line (9). Point (P4) is the node where the waste liquid line (13) branches off from the plasma line (8B) in the extracorporeal circulation system (2). Point (P5) is the node where the regeneration line (15) connects to the connecting line (11) in the extracorporeal circulation system (2). Point (P6) is the node in the extracorporeal circulation system (2) where the bypass line (12) connects to the plasma line (8B).

[0294] According to a preferred embodiment of the present invention, the connecting line (11) is connected to the arterial line (5). According to another preferred embodiment of the present invention, the connecting line (11) is directly connected to the cell separator (7).

[0295] As already described, the apheresis device according to the invention comprises at least one line (so-called Regeneration pipeline(14)), which is capable of supplying a regeneration solution (e.g., an alkali metal hydroxide solution, preferably a sodium hydroxide solution) or a flushing solution or a neutralizing solution into the extracorporeal circulation system, preferably shortly before the single sampling column (4) or directly connected to the single sampling column (4). In this context, it also refers to a regeneration line (14) for connecting at least one liquid container (F) to the extracorporeal circulation system in fluidic connection, that is, liquid from the liquid container can be introduced into the extracorporeal circulation system via the regeneration line.

[0296] According to a preferred embodiment of the present invention, the regeneration line (14) is connected to the plasma line (8A) after point (P2), that is, between point (P2) and the single sampling column (4). According to another preferred embodiment of the present invention, the regeneration line (14) is connected to the plasma line (8A) at point (P2). According to another preferred embodiment of the present invention, the regeneration line (14) is directly connected to the single sampling column (4).

[0297] It is obvious to a person skilled in the art that the liquid containers (F) for connection to the regeneration line itself do not have to be part of the apheresis device according to the invention, since they are usually disposable items, for example in the form of ordinary infusion bags, which are connected to the connecting line by an operator (such as an attending physician or a nurse) according to the specific application.

[0298] According to the invention, there can be a separate regeneration line (14) for connecting a liquid container (F). Here, for example, it is conceivable that a separate liquid container, for example an infusion bag containing an alkali hydroxide solution, preferably a sodium hydroxide solution, can be connected to the regeneration line (14). However, it is also conceivable that the end of the regeneration line (14) to which the liquid container can be connected is located spatially close to the end of the connecting line (11) to which the liquid container can be connected, so that a liquid container (with at least two connection options or a corresponding adapter) can be connected to the connecting line (11) and the regeneration line (14), for example for an infusion bag containing a NaCl solution or a citrate solution.

[0299] According to the invention, the presence of a single regeneration line (14) is possible and 1 or 2 regeneration lines are particularly preferred. In addition, embodiments of the apheresis device according to the invention with two, three or more regeneration lines (14', 14", 14'", etc.) are possible, in which case these two, three or more regeneration lines can flow independently of one another into the extracorporeal circulation system (2) in the region of the branching of the bypass line (12) at the plasma line (8A) (i.e. from point P2) to the apheresis column (4). “Independent of each other” in this article means, for example, that in an embodiment of the single collection device according to the present invention having two regeneration lines (14', 14"), one regeneration line (14') is connected to the plasma line (8A) between point (P2) and the single collection column (4) and the other regeneration line (14") is directly connected to the single collection column (4), but these two regeneration lines (14', 14") can also be connected to the plasma line (8A) between point (P2) and the single collection column (4). One regeneration line (14') can also be connected to the other regeneration line (14"). However, when there are two or more regeneration lines (14', 14", 14'" etc.), it is particularly preferred that all regeneration lines (14', 14", 14'" etc.) are connected to the extracorporeal circulation system (2) at the same point in the area from point (P2) to the single sampling column (4), and it is even more preferred that all regeneration lines (14', 14", 14'" etc.) are connected to the extracorporeal circulation system (2) at point (P2).

[0300] According to the invention, it is particularly advantageous if the connecting line (11) and the regeneration line (14) use the same liquid source, since this not only saves space, but also minimizes the effort required for operation and maintenance of the apheresis device according to the invention. In this way, existing apheresis systems can also be modified or supplemented without the need to connect separate additional large equipment. Therefore, in a preferred embodiment of the invention, the regeneration line (14) branches off from the connecting line (11), wherein the branching point of the regeneration line (14) in the connecting line (11) is referred to as point (P5).

[0301] Therefore, according to some embodiments of the present invention, preferably, at least one regeneration line (14) introduced into the plasma line (8A) or directly introduced into the single sampling column (4) starts from a point (P5) in at least one connecting line (11).

[0302] In an embodiment, where there is more than one connecting line (11', 11", 11"', etc.) and the regeneration line (14) is connected to multiple connecting lines (11', 11", 11"', etc.), the naming of the branch point (P5', P5", P5"', etc.) is based on the naming of the connecting lines (11', 11", 11"', etc.). That is, for example, the regeneration line (14) joins or is connected to two existing connecting lines (11', 11"), the point where the regeneration line (14) joins the connecting line (11') is called point (P5') and the point where the regeneration line (14) joins the connecting line (11") is called point (P5").

[0303] A single collection device (1) is preferred, wherein the single collection device (1) has two connecting pipelines (11', 11"), which respectively connect the liquid containers (F1, F2) to the arterial line (5) or the cell separator (7), wherein two regeneration lines (14', 14") are led out from the two liquid containers (F1, F2) or the two connecting pipelines (11', 11") and introduced into the plasma line (8A) or directly introduced into the single collection column (4).

[0304] Particularly preferred is also an embodiment in which the regeneration line (14) leads into the plasma line (8A) or directly into the apheresis column (4) and, starting from a point (P5) in at least one connecting line (11), has at least one additional connection for a liquid container (see Figure 5 ). An infusion bag containing an alkali metal hydroxide solution, preferably a sodium hydroxide solution, can preferably be connected to this additional connection.

[0305] In a preferred embodiment, the single collection device (I) further comprises at least one regeneration line (14), which is drawn from at least one liquid container (F1) or at least one connecting line (11) and flows in the flow direction at or after the branch of the bypass line (12) to the plasma line (8A) or directly connected to the single collection column (4), wherein at least one regeneration line (14) has at least one additional connection for the liquid container (F2). An infusion bag containing an alkali metal hydroxide solution (preferably a sodium hydroxide solution) can preferably be connected to the additional connection.

[0306] In a preferred embodiment, the apheresis device (I) further comprises at least one second regeneration line (14), which is led out from at least one liquid container (F2) and wherein the second regeneration line (14) has no connection with the arterial line (5) or the cell separator (7) and leads to the plasma line (8A) or directly connects to the apheresis column (4) at or after the branch of the bypass line (12) in the flow direction. An infusion bag with an alkali metal hydroxide solution (preferably a sodium hydroxide solution) can preferably be connected to the second regeneration line (14).

[0307] In the embodiment where there are more connecting lines than regeneration lines, in which each regeneration line establishes a connection with at least one connecting line, it is possible that each regeneration line is connected to one connecting line and the redundant connecting lines are connected only to the arterial line or the cell separator, or that a plurality of connecting lines converge on the regeneration line, i.e. a plurality of connecting lines are connected to one regeneration line. A mixed form is also possible.

[0308] There are various possibilities for regulating the flow rate in the connecting line (11) and in the regeneration line (14) section after point (P5). This can be accomplished, for example, by individually controllable pumps in the connecting line (11) and in the regeneration line (14) section after point (P5). Another possibility is that the pump is located in the connecting line (11) before point (P5), wherein the flow rate distribution after point (P5) is determined by the diameters of the regeneration line (14) and the connecting line (11), or can be regulated by appropriate means (clamps, valves) (e.g. by changing the corresponding line diameters). The regulation of the flow rate is of course particularly important when a solution (e.g. a citrate solution) has to be introduced into the system via the connecting line (11) (e.g. for blood anticoagulation) and has to simultaneously reach the single sampling column via the regeneration line (14) (e.g. as a neutralizing solution). By this mechanism, for example, the solution feed via connecting line (11) can be kept constant (for example for constant anticoagulation) even if the separated solution for neutralizing the single sampling column after regeneration with alkali metal hydroxide solution, preferably sodium hydroxide solution, is branched off via regeneration line (14).

[0309] Compared to other systems, a maximum of 8, preferably 7, further preferably 6, and most preferably 5 pumps are sufficient for the apheresis device (1).

[0310] In an embodiment of the present invention having multiple connecting lines (11', 11", 11"', etc.) and multiple regeneration lines (14', 14", 14"', etc.), it is possible that each connecting line is connected to one of the regeneration lines, which in turn are connected to the plasma line (8A) or directly connected to the single sampling column (4) after point (P2). Here, each regeneration line can be connected to the plasma line (8A) or directly connected to the single sampling column (4) at a point after point (P2) independently of other regeneration lines. However, it is preferred that all regeneration lines are connected to the plasma line (8A) or directly connected to the single sampling column (4) at the same point after point (P2), and it is even more preferred to be directly connected to the single sampling column (4) and most preferably at point (P2). Such an exemplary embodiment can be based on Figure 4To explain: Here, the single collection device (1) has a first connecting line (11'), which firstly leads to the arterial line (5) and secondly branches off a first regeneration line (14') therefrom at point (P5'). The single collection device (1) also has a second connecting line (11"), which firstly leads directly to the cell separator (7) and secondly branches off a second regeneration line (14") therefrom at point (P5"). In this embodiment, the two regeneration lines (14', 14") enter the extracorporeal circulation system (2) at point (P2).

[0311] Therefore, a single collection device (1) is preferred, wherein the single collection device (1) has two connecting lines (11', 11"), respectively connecting at least one liquid container (F) to the arterial line (5) or the cell separator (7), and wherein at least one regeneration line (14) leading to the plasma line (8A) or directly introduced into the single collection column (4) is connected to the connecting line (11') at point (P5') and connected to the connecting line (11") at point (P5").

[0312] Therefore, an embodiment of the apheresis device (1) is particularly preferred, wherein the apheresis device (1) has two connecting lines (11', 11"), connecting at least one liquid container (F1, F2) to the arterial line (5) or the cell separator (7), respectively, and wherein at least one regeneration line (14) introduced into the plasma line (8A) or directly introduced into the apheresis column (4) is connected to the connecting line (11') at point (P5') and connected to the connecting line (11") at point (P5"), and wherein the regeneration line (14') leads from the liquid container (F1) or the connecting line (11') leading from the liquid container (F1) to the apheresis column (4) or to the plasma line (8A) and the regeneration line (14") leads from the liquid container (F2) or the connecting line (11") leading from the liquid container (F2) to the apheresis column (4) or to the plasma line (8A) or leads to the regeneration line (14').

[0313] Therefore, it is particularly preferred that the single collection device (1) has a connecting line (11') for connecting to a liquid container (F1) and a connecting line (11") for connecting to a liquid container (F2), and the connecting line (11') is connected to the arterial line (5) or the cell separator (7), the connecting line (11") is connected to the arterial line (5) or the cell separator (7) or is connected to the connecting line (11') and thus ultimately also connected to the arterial line (5) or to the cell separator (7), and the regeneration line (14') leads from the liquid container (F1) or from the connecting line (11') to the single collection column (4) or to the plasma line (8A), and the regeneration line (14") leads from the liquid container (F2) or from the connecting line (11") to the single collection column (4) or to the plasma line (8A) or is introduced into the regeneration line (14').

[0314] Therefore, particularly preferred is an embodiment of the apheresis device (1), wherein the apheresis device (1) has a connecting line (11') for connecting a liquid container (F1) to an arterial line (5) or a cell separator (7) and a connecting line (11") for connecting a liquid container (F2) to an arterial line (5) or a cell separator (7), and wherein a regeneration line (14') is led out from the liquid container (F1) or the connecting line (11') and introduced into the plasma line (8A) or directly into the apheresis column (4) after branching off from the bypass line (12) in the flow direction, and a regeneration line (14") is led out from the liquid container (F2) or the connecting line (11") and introduced into the plasma line (8A) or into the regeneration line (14') or directly into the apheresis column (4) after branching off from the bypass line (12) in the flow direction.

[0315] According to one embodiment, a preferred apheresis device (1) is one in which the apheresis device (1) has two connecting lines (11', 11"), each for connecting at least one liquid container to the arterial line (5) or the cell separator (7), and in which at least one regeneration line (14) leading to the plasma line (8A) or directly to the apheresis column (4) is connected to the connecting line (11') at point (P5') and to the connecting line (11") at point (P5"). This should be understood as the regeneration line (14) being a connecting element between the connecting line (11', 11") on one side and the plasma line (8A) or the apheresis column (4) on the other side. Therefore, liquid from a liquid container (F) connected to one of the two connecting lines (11', 11") can flow into the plasma line (8A) or directly into the apheresis column (4) via the regeneration line (14) after point (P2).

[0316] The embodiment with two (or even more) connecting pipelines is ideally suitable for regenerating the single sampling column (4) using different regeneration solutions and introducing them successively into the single sampling column (4). For example, such an apparatus is very suitable for first introducing a NaCl solution to replace the plasma contained in the single sampling column, then introducing an alkali metal hydroxide solution (preferably a sodium hydroxide solution) to effectively and quickly regenerate the adsorber, and finally replacing the alkali metal hydroxide solution (preferably a sodium hydroxide solution) contained in the single sampling column with a NaCl solution before the plasma is introduced into the single sampling column again.

[0317] In an embodiment of the single collection device according to the present invention, in which the bypass line (12) leads to a point (P6) in the plasma line (8B), it is preferred that the point (P6) is located before the point (P1) (in the flow direction) (see Figure 1-2 ).

[0318] According to a preferred embodiment of the present invention, the connecting line is connected to the arterial line. According to another preferred embodiment of the present invention, the connecting line is directly connected to the cell separator.

[0319] According to a preferred embodiment of the present invention, the regeneration line (14) is connected to the plasma line (8A) after point (P2), that is, between point (P2) and the single sampling column (4). According to another preferred embodiment of the present invention, the regeneration line (14) is directly connected to the single sampling column (4).

[0320] In order to reduce the dead volume of the system, it is particularly preferred according to the invention that in the apheresis device (1) according to the invention at least one regeneration line (14) is connected to the extracorporeal circulation system (2) at point (P2). In embodiments in which more than one regeneration line (14', 14", 14'" etc.) is present, it is particularly preferred that all existing regeneration lines (14', 14", 14'" etc.) are connected to the extracorporeal circulation system (2) at point (P2) or to the plasma line (8A) at point (P2).

[0321] Therefore, the present invention also relates to an apheresis device (1) according to the present invention, wherein a bypass line (12) leads from a point (P2) in the plasma line (8A) to a point (P6) in the plasma line (8B). A waste liquid line (13) leads from a point (P4) in the plasma line (8B), and at least one regeneration line (14) leads into the plasma line (8A) at point (P2).

[0322] In order to further reduce the dead volume of the system, it is even more preferred that not only the regeneration line (14) joins the plasma line (8A) at point (P2) and the bypass line (12) also branches off from the plasma line (8A) at that point, but also the waste line (13) also joins the bypass line (12) at the same point where it branches off from the plasma line (8B). In other words, it is preferred that the point (P6) where the bypass line (12) joins the plasma line (8B) coincides with the point (P4) where the waste line (13) branches off from the plasma line (8B), i.e., when P4=P6 (see also Figure 2 ).

[0323] Therefore, the present invention also relates to a single collection device (1) according to the present invention, wherein a bypass line (12) leads from a point (P2) in a plasma line (8A) to a point (P6) in a plasma line (8B), a waste liquid line (13) leads from a point (P4) in a plasma line (8B), and at least one regeneration line (14) is connected to the plasma line (8A) at a point (P2), and wherein point (P6) is identical to point (P4).

[0324] In the device according to the invention, a cell separator is installed, which separates the patient's blood supplied to it (via the arterial line) into plasma and cellular components and further transmits these parts via corresponding lines (i.e., plasma line and cell line). As already mentioned, it must be taken into account that the separation of plasma and cellular components by the cell separator used is not complete, but preferably only 10% to 90% of the total plasma is separated from the cellular components. In the case of using a centrifugal cell separator, preferably 70% to 90%, more preferably 80% to 87% of the total plasma is separated from the cellular components. In the case of using a membrane cell separator, preferably 10% to 30%, more preferably 13% to 25%, and even more preferably 15% to 20% of the total plasma is separated from the cellular components.

[0325] Possible types of cell separators that can be used in connection with the present invention include centrifugal cell separators, membrane cell separators, such as membrane cell separators with a semipermeable membrane and membrane cell separators with a rotating membrane.

[0326] Therefore, the present invention also relates to an apheresis device for removing CRP from blood in vitro, wherein the cell separator (7) is a centrifugal cell separator or a membrane cell separator.

[0327] In the present application, the position of one or more components of the apheresis device according to the invention relative to another component of the apheresis device according to the invention is indicated by the terms "before" or "after" (or "flow direction . . . before" and "flow direction . . . after"), which refers to the general flow direction of blood or plasma in the apheresis device according to the invention. Therefore, "before" in relation to a component of the device according to the invention means against the general flow direction of blood or plasma, and "after" with respect to a component of the device according to the invention thus means in the general flow direction of blood or plasma. Preferably, the flow direction in the apheresis device is not reversed or is not reversed by the device used to generate and regulate the flow.

[0328] According to the present invention, the apheresis device for extracorporeal removal of CRP from blood comprises an apheresis column (4) for affinity chromatography removal of CRP from blood or plasma, the function of which is to bind CRP present in the patient's blood or plasma and conducted through the apheresis column (4).

[0329] Pump

[0330] According to the invention, in the apheresis device according to the invention for the extracorporeal removal of CRP from blood, a device for generating and regulating the flow of blood (or plasma) in the extracorporeal circulation system is provided. To this end, one or more pumps or pump systems are usually used, which enable the blood (or plasma or also regeneration solution or anticoagulant solution) to flow controllably through the extracorporeal circulation system and can fluidically connect the components of the device according to the invention.

[0331] According to the invention, the preferred flow direction within the extracorporeal circulation system and the components of the device according to the invention fluidically connected thereto is from a channel in the patient through which blood enters the device according to the invention, through the arterial line of the extracorporeal circulation system to the venous line of the extracorporeal circulation system and to a channel in the patient through which the treated blood is returned to the patient.

[0332] The device for generating and regulating flow according to the present invention used in the extracorporeal circulation system is preferably a peristaltic pump (also called a hose pump), a piston pump, a pneumatic pump, a hydraulic pump or other pump types known to those skilled in the art. Therefore, the term "device for generating and regulating flow" and the term "pump" can be used as synonyms in this article.

[0333] According to the invention, the device used in the extracorporeal circulation system for generating and regulating the flow of blood (or plasma or regeneration solution or anticoagulant solution) preferably has no direct physical contact with the blood (or plasma or regeneration solution or anticoagulant solution) in the device according to the invention. This is particularly advantageous for hygienic reasons and prevents contamination of complex mechanical parts such as pumps with blood and, of course, also prevents contamination of blood with the device used for generating the flow.

[0334] In a particularly preferred embodiment of the invention, the means for generating and regulating the flow in the extracorporeal circulation system are therefore one or more peristaltic pumps.

[0335] The exact location of the device for generating and regulating the flow in the extracorporeal circulation system (i.e. one or more pumps) is not essential for the present invention. An embodiment of the present invention using only one pump is possible, wherein the pump is located in the area of ​​the arterial line of the apheresis device for extracorporeal removal of CRP from blood according to the present invention, i.e. before the cell separator. According to the present invention, if a plurality of devices for generating and regulating the flow in the extracorporeal circulation system, i.e. several pumps, are provided, then preferably these devices can be controlled and regulated independently of each other (e.g. by a CPU; e.g. by a central processing unit). Different flow rates within the extracorporeal circulation system may be desired or required, respectively, depending on the specific application. It is also conceivable that different flow rates are required in different components of the device according to the present invention during a specific application.

[0336] According to the present invention, multiple devices (i.e. pumps) for generating and regulating flow can also be integrated in the apheresis device according to the present invention. Therefore, the device for generating and regulating flow can be located in the arterial line (5) and / or in the plasma line (8A) and / or in the plasma line (8B) and / or in the venous line (6) and / or in the bypass line (12) and / or in the cell line (9) and / or in the connecting line (11) and / or in the connecting line (11', 11", 11"', etc.) and / or in the regeneration line (14) and / or in the regeneration line (14', 14", 14"', etc.). As described above, according to one embodiment of the present invention, in which the regeneration line (14) branches off from the connecting line (11) at point (P5), the device for generating and regulating the (inorganic salt solution) flow is preferably placed in the connecting line (11) before point (P5).

[0337] According to another embodiment of the present invention, in which the regeneration line (14) branches off from the connecting line (11) at point (P5), preferably, a device for generating and regulating a flow rate is disposed in the connecting line (11) after point (P5) and a device for generating and regulating a flow rate is disposed in the regeneration line (14).

[0338] Furthermore, the apheresis device (1) preferably has at least one particle filter which is arranged in the plasma line (8B) or the venous line (6) downstream of the apheresis column (4) in the flow direction.

[0339] Furthermore, the apheresis device (1) preferably has at least one bubble trap which is arranged in the plasma line (8B) or the venous line (6) downstream of the apheresis column (4) in the flow direction.

[0340] In the case of a centrifuge as cell separator (7), the apheresis device (1) preferably has at least one plasma container which is arranged in the plasma line (8A) downstream of the centrifuge (7) and upstream of the apheresis column (4) in the flow direction.

[0341] In a further embodiment, the apheresis device for extracorporeal removal of CRP from blood or plasma according to the present invention may include one or more Pressure Sensors, which is used to measure or monitor the pressure in a specific part of the device according to the invention. This is not only used to monitor and adjust the operating parameters of the apheresis device according to the invention, but also advantageously in the event of a malfunction (for example, a tube or filter of the device is clogged) the operation can be stopped to avoid adverse consequences for the patient. The exact operating modes and installation positions in the device according to the invention are part of the prior art and are known to the person skilled in the art. In a preferred embodiment of the invention, at least one pressure sensor is arranged in the arterial line of the apheresis device according to the invention and at least one pressure sensor is arranged in the venous line of the apheresis device according to the invention. In another preferred embodiment of the invention, such a pressure sensor is integrated in a device for generating and regulating the flow in the extracorporeal circulation system for the apheresis device according to the invention.

[0342] The pressure or pressure range of the NaOH solution is preferably from -200 to 200 mbar; pressure difference; before and after the adsorber: maximum 300 mbar.

[0343] In order to be able to control the flow direction in the system at the node of the extracorporeal circulation system, that is, at the point where multiple pipelines converge or fork each other, a device for determining the flow of a solution (e.g., blood, plasma, or regeneration solution) is preferably provided. This can be a valve, a multi-way valve, a clamp, or a stop valve, a check valve, a pressure valve, a directional valve, or other valve types known to those skilled in the art, which allow flow in a specific direction and prevent another direction. Preferably, this device (e.g., a valve) for regulating flow is located at point (P1) and / or point (P2) and / or point (P3) and / or point (P4) and / or point (P5) and / or at point (P6). In addition, for example, it is possible to connect two or more valves in series to achieve more complex flow regulation.

[0344] It is also particularly preferred that the means for regulating the flow (eg valves) can be controlled electronically, ie their operation can be effected by a central processing unit.

[0345] Therefore, the present invention also relates to an apheresis device for extracorporeal removal of CRP from blood, wherein electronically controlled valves are provided at points (P1), (P2), (P4), (P5), (P6), (P7) and (P8).

[0346] According to the invention, it is also conceivable that the valves are not located directly at the branching points (P1, P2, P4, P5, P6, P7 and P8), but are located in the upstream and / or downstream lines and control the flow of the solution in the extracorporeal circulation system in this way. Hose clamps can also be used for this purpose. It is particularly preferred that these valves or hose clamps are electronically controlled.

[0347] Another advantage of the present invention is that, in connection with the fact that apheresis and regeneration of the apheresis column are realized in a single device, the entire device can be controlled by a single central processing unit (CPU). Therefore, different items in the apheresis process, such as the normal operation of passing plasma through the apheresis column, and the regeneration operation of draining plasma from the apheresis column through a bypass line and flushing the apheresis column with a regeneration solution, can be controlled by a single processing unit or the software therein. This promotes the automation of many processes, thereby reducing the room for human operating errors. On the other hand, in the devices of the prior art, different complex systems must be combined (a primary system for separating blood into plasma and cellular components; and a secondary system for apheresis and regeneration), each of which must be controlled separately.

[0348] Therefore, the present invention also relates to an apheresis device for the extracorporeal removal of CRP from blood, wherein the entire device is controlled by only one central processing unit.

[0349] Another aspect of the present invention relates to an apheresis device (II) for extracorporeal removal of CRP from a patient's blood, wherein the apheresis device comprises at least two apheresis columns, and wherein the apheresis device (II) is configured to withstand an alkaline hydroxide solution (preferably a sodium hydroxide solution).

[0350] In other words, another aspect of the present invention relates to an apheresis device (II) for extracorporeal removal of CRP from the blood of a patient, wherein the apheresis device comprises at least two apheresis columns, wherein the apheresis device can be connected to the patient's blood circulation, and wherein the apheresis device (II) is designed to be resistant to alkali metal hydroxide solutions, preferably sodium hydroxide solutions.

[0351] Another aspect of the present invention relates to an apheresis device, wherein at least one additional apheresis column (4"), preferably a second apheresis column (4"), is connected to a bypass line, or the bypass line comprises at least one additional apheresis column (4"), preferably a second apheresis column. Preferably, at least one additional apheresis column (4"), preferably a second apheresis column (4"), is included in the bypass line. Therefore, the apheresis device according to the present invention for extracorporeal removal of CRP from blood as described herein may comprise at least one additional apheresis column (4"), preferably a second apheresis column (4"), wherein the at least one additional apheresis column (4"), preferably a second apheresis column (4"), is included in the bypass line. When a section of the bypass line (12') is connected to the second apheresis column (4") and another section of the bypass line (12") is led out at the outlet of the apheresis column (4"), the apheresis column (4") is included in the bypass line.

[0352] Therefore, another aspect of the present invention is an apheresis device (II) for extracorporeal removal of CRP from blood, comprising:

[0353] Extracorporeal circulation system for blood (2),

[0354] Device (3) for generating and regulating blood flow in an extracorporeal circulation system (2),

[0355] a cell separator (7) for separating blood into plasma and cellular components,

[0356] Two single sampling columns (4', 4") for affinity chromatography removal of CRP from plasma,

[0357] The extracorporeal circulation system (2) comprises an arterial line (5) to a cell separator (7), a plasma line (8A) from the cell separator (7) to a single collection column (4'), a plasma line (8B) for CRP-depleted plasma from the single collection column (4') to a node (P1), a cell line (9) for separated cellular components from the cell separator (7) to the node (P1), and a venous line (6) from the node (P1).

[0358] at least one connecting line (11) for connecting at least one liquid container (F) to the arterial line (5) or to the cell separator (7),

[0359] It is characterized in that

[0360] A bypass line (12) is branched from the plasma line (8A) and connected to the plasma line (8B), and the bypass line (12) includes a second single sampling column (4").

[0361] The waste liquid pipeline (13) is led directly from the single collection column (4') or from the plasma pipeline (8B) before the confluence of the bypass pipeline (12) in the flow direction, and

[0362] At least one regeneration line (14) leads to the plasma line (8A) or directly connects to the single sampling column (4') after the branch of the bypass line (12) in the flow direction, and

[0363] The second single sampling column (4") is connected in parallel with the first single sampling column (4'), and the two single sampling columns (4', 4") can only be operated alternately, that is, they cannot be used to remove CRP at the same time, wherein the single sampling device (II) is designed to be tolerant to alkali metal hydroxide solution (preferably sodium hydroxide solution).

[0364] Preferably, the single sampling device (II) further comprises a central processing unit for controlling the single sampling device (II).

[0365] Therefore, another aspect of the present invention is an apheresis device (II) for extracorporeal removal of CRP from blood, comprising:

[0366] Extracorporeal circulation system for blood (2),

[0367] Device (3) for generating and regulating blood flow in an extracorporeal circulation system (2),

[0368] a cell separator (7) for separating blood into plasma and cellular components,

[0369] Two single sampling columns (4', 4") for affinity chromatography removal of CRP from plasma,

[0370] The extracorporeal circulation system (2) comprises an arterial line (5) to a cell separator (7), a plasma line (8A) from the cell separator (7) to a single collection column (4'), a plasma line (8B) for CRP-depleted plasma from the single collection column (4') to a node (P1), a cell line (9) for separated cellular components from the cell separator (7) to the node (P1), and a venous line (6) from the node (P1).

[0371] at least one connecting line (11) for connecting at least one liquid container (F1) to the arterial line (5) or to the cell separator (7),

[0372] It is characterized in that

[0373] A bypass line (12) is branched from the plasma line (8A) and connected to the plasma line (8B), and the bypass line (12) includes a second single sampling column (4").

[0374] The waste liquid pipeline (13) is led directly from the single collection column (4') or from the plasma pipeline (8B) before the confluence of the bypass pipeline (12) in the flow direction, and

[0375] at least one regeneration pipeline (14), which is led out from at least one liquid container (F1) or at least one connecting pipeline (11) and is led to the plasma pipeline (8A) or directly connected to the single sampling column (4') at or after the branch of the bypass pipeline (12) in the flow direction, and

[0376] at least a second regeneration line (14) which leads from at least one liquid container (F2), wherein the second regeneration line (14) is not connected to the arterial line (5) or the cell separator (7) and leads in the flow direction to the plasma line (8A) or directly to the single sampling column (4') at or after the branch of the bypass line (12),

[0377] The second single sampling column (4") is connected in parallel with the first single sampling column (4'), and the two single sampling columns (4', 4") can only be operated alternately, that is, they cannot be used to remove CRP at the same time, wherein the single sampling device (II) is designed to be tolerant to alkali metal hydroxide solution (preferably sodium hydroxide solution).

[0378] Preferably, the single sampling device (II) further comprises a central processing unit for controlling the single sampling device (II).

[0379] Preferably, the single collection device (II) therefore comprises at least two regeneration lines (14', 14") which are independently led in the flow direction at or after the branch of the bypass line (12) to the plasma line (8A) or directly connected to the single collection column (4'), wherein at least one of the regeneration lines (14', 14") is led out from at least one liquid container (F1) or at least one connecting line (11).

[0380] Therefore, another aspect of the present invention is an apheresis device (II) for extracorporeal removal of CRP from blood, comprising:

[0381] Extracorporeal circulation system for blood (2),

[0382] Device (3) for generating and regulating blood flow in an extracorporeal circulation system (2),

[0383] a cell separator (7) for separating blood into plasma and cellular components,

[0384] Two single sampling columns (4', 4") for affinity chromatography removal of CRP from plasma,

[0385] The extracorporeal circulation system (2) comprises an arterial line (5) to a cell separator (7), a plasma line (8A) from the cell separator (7) to a single collection column (4'), a plasma line (8B) for CRP-depleted plasma from the single collection column (4') to a node (P1), a cell line (9) for separated cellular components from the cell separator (7) to the node (P1), and a venous line (6) from the node (P1).

[0386] at least one connecting line (11) for connecting at least one liquid container (F1) to the arterial line (5) or to the cell separator (7),

[0387] It is characterized in that

[0388] A bypass line (12) is branched from the plasma line (8A) and connected to the plasma line (8B), and the bypass line (12) includes a second single sampling column (4").

[0389] The waste liquid pipeline (13) is led directly from the single collection column (4') or from the plasma pipeline (8B) before the confluence of the bypass pipeline (12) in the flow direction, and

[0390] At least one regeneration line (14) is led out from at least one liquid container (F1) or at least one connecting line (11) and is led in the flow direction at or after the branch of the bypass line (12) to the plasma line (8A) or directly connected to the single sampling column (4'), wherein at least one regeneration line (14) has at least one additional connection for the liquid container (F2),

[0391] The second single sampling column (4") is connected in parallel with the first single sampling column (4'), and the two single sampling columns (4', 4") can only be operated alternately, that is, they cannot be used to remove CRP at the same time, wherein the single sampling device (II) is designed to be tolerant to alkali metal hydroxide solution (preferably sodium hydroxide solution).

[0392] Preferably, the apheresis device (II) further comprises a central computing unit (10) for controlling the apheresis device (II).

[0393] The above-described embodiment of the apheresis device (1) according to the invention is transferred to the apheresis device (II) according to the invention. The bypass line (12) of the apheresis device (1) is used as the plasma line in the apheresis device (II).

[0394] With the aid of this single sampling device (II) according to the present invention, CRP can be removed from the blood more effectively within the same treatment time compared to the prior art devices. By using two single sampling columns in parallel, which can only be used alternately for the removal of CRP, one single sampling column can be used to remove CRP from the blood by the single sampling device according to the present invention, and the second single sampling column can be replaced with another single sampling column, or the second single sampling column can be regenerated in the ongoing single sampling treatment. Therefore, high clinical throughput can also be achieved with the aid of the single sampling device. In addition, the use of the single sampling device according to the present invention is not limited by the dead volume. Generally, oversized single sampling columns and single sampling columns connected in series severely limit their use for single sampling due to their large dead volume. In addition, the capacity of the single sampling device and the capacity or number of single sampling columns connected in series are limited by the blood flow rate of the human body. In addition, due to the large dead volume, even a single sampling device with single sampling columns connected in parallel and used simultaneously cannot be used to remove CRP from the blood effectively and without risk to the patient.

[0395] As described herein, the single sampling device (II) according to the present invention is characterized in that the second single sampling column is connected in parallel with the first single sampling column (4'). "Parallel" herein refers to multiple circulations that exist adjacent to each other in the extracorporeal circulation system (2), that is, for example, the first single sampling column (4') and the plasma line (8A) for separating plasma and the plasma line (8B) for plasma depleted in CRP constitute the first circulation system of the extracorporeal circulation system (2), and the second single sampling column (4") and the bypass line segment (12') of the bypass line (12) and the bypass line segment (12") of the bypass line (12) constitute the second circulation system of the extracorporeal circulation system (2). "Parallel" also means that the two single sampling columns are not connected in series, that is, not one after another, so that the effluent of the first single sampling column is introduced into the second single sampling column. Due to the parallel arrangement of the single sampling columns, their capacities will not be added.

[0396] Different from this is the serial connection of the single collection columns, which is not according to the present invention. "Serial connection" means that multiple single collection columns are present in only one cycle of the extracorporeal circulation system (2), that is, for example, the first single collection column (4') and the second single collection column (4") together with the plasma pipeline (8A) and the plasma pipeline (8B) only form one cycle of the extracorporeal circulation system (2), that is, they are connected or arranged in series.

[0397] According to the present invention, the two single sampling columns (4', 4") connected in parallel or arranged in parallel can only be operated alternately. "Alternating" means that the separated plasma passes through the single sampling column (4') or through the single sampling column (4") but not through both single sampling columns (4', 4") at the same time. The "alternating" operation in this context refers to the therapeutic removal of CRP. The two single sampling columns (4' and 4") cannot be used to remove CRP at the same time. Of course, one of the two single sampling columns can be regenerated and the other can be used for CRP removal at the same time. Only the simultaneous therapeutic operation of the two single sampling columns for removing CRP is excluded.

[0398] The following situation is thus possible: plasma is passed through one apheresis column to remove CRP. At the same time, a second apheresis column is ready for use and the plasma can be diverted to this second apheresis column. Once the capacity of the first apheresis column is exhausted or other problems occur in the first apheresis column or the second apheresis column has been used for CRP removal and must be replaced or regenerated, or the second apheresis column is regenerated during the removal of CRP by the first apheresis column.

[0399] In an embodiment of the present invention, the apheresis device (II) having two apheresis columns is therefore configured so that the apheresis columns can only be operated alternately.

[0400] Therefore, according to one embodiment of the apheresis device (II) according to the present invention, plasma can pass only through the first apheresis column (4') or only through the second apheresis column (4") at the same time. In a further embodiment of the device according to the present invention, the apheresis device is therefore designed so that plasma can pass only through the first apheresis column (4') or only through the second apheresis column (4") at the same time.

[0401] During the alternating operation of the two single sampling columns (4', 4"), either no plasma passes through the single sampling column (4') or no plasma passes through the single sampling column (4"). This provides the possibility of replacing one of the two single sampling columns from the single sampling device during operation of the single sampling device. "Replacement" in this context means replacing one of the two single sampling columns with a new one or regenerating one of the two single sampling columns. Regeneration of one of the two single sampling columns can be carried out, for example, by flushing with an alkali metal hydroxide solution (preferably sodium hydroxide solution). For the regeneration of the single sampling column, an alkali metal hydroxide solution (preferably sodium hydroxide solution) is preferably used. In this context, "during operation" means that the process of removing CRP from the blood is still ongoing.

[0402] Therefore, as described herein, one embodiment of the single sampling device (II) according to the present invention relates to a single sampling device, wherein the first single sampling column (4') is replaceable during the operation of the second single sampling column (4"), and the second single sampling column (4") is replaceable during the operation of the first single sampling column (4').

[0403] Also conceivable are embodiments in which the first apheresis column (4') is regenerable during operation of the second apheresis column (4") and the second apheresis column (4") is regenerable during operation of the first apheresis column (4').

[0404] Therefore, in one embodiment of the present invention, the single sampling device (II) is designed so that the first single sampling column (4') is replaceable or regenerable in the second single sampling column (4") and the second single sampling column (4") is replaceable or regenerable during the operation of the first single sampling column (4').

[0405] Also preferred is an embodiment of an apheresis device (II) for extracorporeal removal of CRP from blood, wherein the second apheresis column (4") is connected in parallel to the first apheresis column (4'), and the two apheresis columns (4', 4") can only be operated alternately at the same time, and wherein the first apheresis column (4') is replaceable or regenerable during the operation of the second apheresis column (4"), and the second apheresis column (4") is replaceable or regenerable during the operation of the first apheresis column (4'), wherein the apheresis device (II) is designed to be resistant to alkali metal hydroxide solutions (preferably sodium hydroxide solutions).

[0406] Also preferred is an embodiment of the apheresis device (II) for extracorporeal removal of CRP from blood, wherein the second apheresis column (4") is connected in parallel with the first apheresis column (4') and the two apheresis columns (4', 4") cannot be used for the removal of CRP at the same time, and wherein one of the apheresis columns (4', 4") can be regenerated for the removal of CRP by another apheresis column at the same time, wherein the apheresis device (II) is designed to be resistant to alkali metal hydroxide solutions (preferably sodium hydroxide solutions).

[0407] Therefore, according to one embodiment of the present invention, the single sampling device (II) is configured so that the first single sampling column (4') is replaceable during the operation of the second single sampling column (4"), and the single sampling column is designed so that it is regenerable, and the second single sampling column (4") is replaceable during the operation of the first single sampling column (4'), and is designed so that the single sampling column (4") is regenerable.

[0408] The second single sampling column (4") connected in parallel with the first single sampling column (4') can be incorporated into the bypass pipeline, that is, the bypass pipeline (12) is composed of a bypass pipeline section (12') and a bypass pipeline section (12"), wherein the second single sampling column (4") is located between the bypass pipeline sections. Therefore, a preferred single sampling device (II) is characterized in that the bypass pipeline section (12') of the bypass pipeline (12) branches off from the plasma pipeline (8A) and is connected to the second single sampling column (4'), and the bypass pipeline section (12") of the bypass pipeline (12") is led out from the single sampling column (4") and connected to the plasma pipeline (8B).

[0409] Also preferred is an embodiment of the apheresis device (II) for extracorporeal removal of CRP from blood,

[0410] The second single sampling column (4") is connected in parallel with the first single sampling column (4'), and the two single sampling columns (4', 4") cannot be used to remove CRP at the same time (i.e., they can only be operated alternately), wherein the single sampling device (1) is designed to be tolerant to alkali metal hydroxide solutions (preferably sodium hydroxide solutions).

[0411] According to one embodiment of the present invention, it is particularly preferred that the single collection device (II) according to the present invention has two connecting lines (11', 11"), each connected to at least one liquid container (F), wherein the connecting lines (11', 11") are independently connected to the arterial line (5) or directly connected to the cell separator (7). Therefore, both connecting lines (11', 11") are connected to the arterial line (5), or both connecting lines (11', 11") are directly connected to the cell separator (7), or it is particularly preferred that one connecting line (11') is connected to the arterial line (5) and the other connecting line (11") is directly connected to the cell separator (7). This allows the two connecting lines (11', 11") to be connected to different liquid containers.

[0412] According to another embodiment of the present invention, the single collection device (II) comprises a waste liquid pipeline (13'), which is directly led out from the single collection column (4') or led out from the plasma line (8B) along the flow direction before the bypass line segment (12") of the bypass line merges into the plasma line (8B) and a waste liquid pipeline (13"), which is directly led out from the single collection column (4") or led out from the bypass line segment (12") along the flow direction before merging into the plasma line (8B).

[0413] Therefore, the present invention also relates to an apheresis device (II) for extracorporeal removal of CRP from blood, comprising:

[0414] Extracorporeal circulation system for blood (2),

[0415] Device (3) for generating and regulating blood flow in an extracorporeal circulation system (2)

[0416] a cell separator (7) for separating blood into plasma and cellular components,

[0417] Two single sampling columns (4', 4") for affinity chromatography removal of CRP from plasma,

[0418] The extracorporeal circulation system (2) comprises an arterial line (5) to a cell separator (7), a plasma line (8A) from the cell separator (7) to a single collection column (4'), and a plasma line (8B) from the single collection column (4') to a point (P1) for plasma depleted of CRP.

[0419] a cell line (9) for separating cell components from the cell separator (7) to the point (P1), and a venous line (6) starting from the point (P1),

[0420] at least one connecting line (11) for connecting at least one liquid container (F) to the arterial line (5) or to the cell separator (7),

[0421] It is characterized in that

[0422] The bypass line section (12') of the bypass line (12) branches off from the plasma line (8A) and is connected to the second single sampling column (4'), and the bypass line section (12") of the bypass line (12") starts from the single sampling column (4") and is connected to the plasma line (8B).

[0423] The waste liquid pipeline (13') is directly led out from the single sampling column (4') or is led out from the plasma pipeline (8B) before the confluence of the bypass pipeline section (12") of the bypass pipeline (12) in the flow direction, the waste liquid pipeline (13") is directly led out from the single sampling column (4") or is led out from the plasma pipeline (8B) before the confluence of the bypass pipeline section (12") of the bypass pipeline in the flow direction, and the waste liquid pipeline (13") is directly led out from the single sampling column (4") or is led out from the bypass pipeline section (12") before entering the confluence of the plasma pipeline (8B) in the flow direction,

[0424] and

[0425] at least one regeneration pipeline (14), which is connected to the plasma pipeline (8A) after the branch of the bypass pipeline section (12') of the bypass pipeline (12) in the flow direction, or is led to the bypass pipeline section (12') or is directly connected to the single sampling column (4') or the single sampling column (4"), and

[0426] The second single sampling column (4") is connected in parallel with the first single sampling column (4'), and the two single sampling columns (4', 4") cannot be used for removing CRP at the same time, that is, they can only be operated alternately, wherein the single sampling device (II) is designed to be tolerant to alkaline hydroxide solution (preferably sodium hydroxide solution).

[0427] According to another embodiment of the present invention, the single sampling device (II) further comprises at least one regeneration line (14), which is led out from at least one liquid container (F) or at least one connecting line (11), and is introduced into the plasma line (8A) or the bypass line segment (12') of the bypass line (12), or is directly connected to the single sampling column (4') or is directly connected to the single sampling column (4"). According to another embodiment of the present invention, the single sampling device (II) further comprises at least one regeneration line (14), which is connected to the bypass line segment (12') in the area between point (P2) and the single sampling column (4'), or is connected to the plasma line (8A) or is directly connected to the single sampling column (4') or is directly connected to the single sampling column (4") in the area between point (P2) and the single sampling column (4").

[0428] In a particularly preferred embodiment of the present invention, the single sampling device (II) includes a waste liquid pipeline (13'), which is directly led out from the single sampling column (4') or led out from the plasma line (8B) before the confluence of the bypass line segment (12") of the bypass line (12) in the flow direction, and at least one regeneration line (14), which is led out from at least one liquid container (F) or at least one connecting line (11) and leads to the bypass line segment (12') or connected to the plasma line (8A) or directly connected to the single sampling column (4') or directly connected to the single sampling column (4").

[0429] In a particularly preferred embodiment of the present invention, the single sampling device (II) includes a waste liquid pipeline (13'), which is directly led out from the single sampling column (4') or led out from the plasma line (8B) before the confluence of the bypass line segment (12") of the bypass line in the flow direction, a waste liquid pipeline (13"), which is directly led out from the single sampling column (4") or led out from the bypass line segment (12") of the bypass line before the confluence of the bypass line segment (12') of the bypass line in the flow direction, and at least one regeneration pipeline (14), which is led out from at least one liquid container (F) or at least one connecting pipeline (11) and led to the bypass line segment (12') of the plasma line (8A) or the bypass line (12) or directly introduced into the single sampling column (4') or directly connected to the single sampling column (4").

[0430] Furthermore, an embodiment of the apheresis device (II) according to the invention is possible in which at least one regeneration line (14) leads to point (P7) and line (15') leads from point (P7) to point (P2) or to plasma line (8A), and line (15") connects from point (P7) to plasma line (8A) (see Figure 7 ).

[0431] In the case of a plasma line (8A) between at least one regeneration line (14) access point (P2) for flushing solution and the single sampling column (4'), or in the case of a bypass line segment (12') of a bypass line (12) between at least one regeneration line (14) access point (P2) and the single sampling column (4"), the flushing solution can be used only for the single sampling column (4') or only for the single sampling column (4"). The regeneration line (14) is therefore either selective for the single sampling column (4') or selective for the single sampling column (4").

[0432] Similarly, an embodiment of the single sampling device (II) according to the present invention having two, three or more regeneration lines (14', 14", 14'", etc.) is possible, wherein these two, three or more regeneration lines can be independently connected to the plasma line (8A) [i.e., from point (P2) to the single sampling column (4')] or to the bypass line segment (12') [i.e., from point (P2) to the single sampling column (4")] or to the single sampling column (4') or to the single sampling column (4"). “Independent of each other” in this article first refers to that in an embodiment of the single collection device according to the present invention having two regeneration lines (14', 14"), one regeneration line (14') is connected to the plasma line (8A) between point (P2) and the single collection column (4'), and the other regeneration line (14") is directly connected to the single collection column (4"), but these two regeneration lines (14', 14") can also be connected to the plasma line (8A) between point (P2) and the single collection column (4'). Another possibility is that one regeneration line (14') is connected to the extracorporeal circulation system (2) at point (P2), and the other regeneration line (14") is connected to the bypass line section (12') of the bypass line (12) between point (P2) and the single collection column (4"). It is also conceivable that one regeneration line (14') is connected to the extracorporeal circulation system at point (P2), and the other regeneration line (14") is connected to the single collection column (4"). The regeneration line (14') can also be connected to another regeneration line (14"). However, when there are two or more regeneration lines (14', 14", 14'", etc.), preferably all regeneration lines (14', 14", 14'", etc.) enter the extracorporeal circulation system (2) at point (P2), wherein the single collection device (II) is designed to be resistant to alkali metal hydroxide solution (preferably sodium hydroxide solution).

[0433] It is further preferred that one regeneration line (14') is connected to the bypass line segment (12') of the bypass line (12) between point (P2) and the single sampling column (4') and another regeneration line (14") is connected to the bypass line segment (12') of the bypass line (12) between point P2 and the single sampling column (4"). It is more preferred that the regeneration line (14') is connected to the single sampling column (4') and the other regeneration line (14") is connected to the single sampling column (4"). Here, the regeneration line (14') is selective for the first single sampling column (4'), while the regeneration line is selective for the second single sampling column (4").

[0434] According to a particularly preferred embodiment of the present invention, the single sampling device (II) therefore also includes a regeneration pipeline (14') for a selective flushing solution for the first single sampling column (4'), and / or further includes a regeneration pipeline (14") for a selective flushing solution for the second single sampling column (4").

[0435] As described above, the regeneration solution required for regenerating the single sampling column can be fed into the extracorporeal circulation system (2) via the regeneration pipeline (14), thereby using a regeneration solution (such as an alkali metal hydroxide solution, preferably a sodium hydroxide solution) in addition to the flushing solution. The flushing solution can but does not have to be used to regenerate the first single sampling column (4') and / or the single sampling column (4"), but in addition to the above functions, its task is to replace the plasma in the regional pipeline (8A) from point (P2) to the single sampling column (4') and the plasma in the plasma pipeline (8B) from the single sampling column (4') to point (P8) or the plasma in the bypass pipeline segment (12') of the bypass pipeline (12) from point (P2) to the region of the blood component column (4") and the plasma in the bypass pipeline segment (12") of the bypass pipeline (12) from the single sampling column (4") to point (P8) before using the regeneration solution, and the flushing solution is discarded via the waste liquid pipeline (13', 13") after passing through one of the two single sampling columns (4', 4").

[0436] It can be envisaged that the single sampling columns (4', 4") connected in parallel can not only be operated alternately, but also be regenerated alternately.

[0437] As described herein, according to the single sampling device (II) of the present invention, during the operation of the second single sampling column (4"), the first single sampling column (4') can be replaceable or regenerated, and during the operation of the first single sampling column (4'), the second single sampling column (4") can be replaceable or regenerated.

[0438] In other words, a single sampling device (II) is preferred, wherein the second single sampling column (4") is connected in parallel with the first single sampling column (4'), and the two single sampling columns (4', 4") can only be operated alternately, and wherein the first single sampling column (4') is replaceable or regenerated during the operation of the second single sampling column (4"), and the second single sampling column (4") is replaceable or regenerated during the operation of the first single sampling column (4'), wherein the single sampling device (II) is designed to be resistant to alkali metal hydroxide solutions (preferably sodium hydroxide solutions).

[0439] Furthermore, an embodiment of the present invention can be envisaged in which the single sampling device has a regeneration line (14) for each liquid container (F), which regeneration line leads from the respective liquid container (F) or their connecting line (11) and is respectively introduced into the plasma line (8A) or into the bypass line segment (12') of the bypass line (12) or directly into the single sampling column (4') or directly into the single sampling column (4").

[0440] A particularly preferred embodiment of the present invention relates to an apheresis device (II) for extracorporeal removal of CRP from blood, comprising:

[0441] Extracorporeal circulation system for blood (2),

[0442] Device (3) for generating and regulating blood flow in an extracorporeal circulation system (2)

[0443] a cell separator (7) for separating blood into plasma and cellular components,

[0444] Two single sampling columns (4', 4") for affinity chromatography removal of CRP from plasma,

[0445] The extracorporeal circulation system (2) comprises an arterial line (5) to a cell separator (7), a plasma line (8A) from the cell separator (7) to a single collection column (4'), and a plasma line (8B) from the single collection column (4') to a point (P1) for plasma depleted of CRP.

[0446] a cell line (9) for separating cell components from the cell separator (7) to the point (P1), and a venous line (6) starting from the point (P1),

[0447] at least one connecting line (11) for connecting at least one liquid container (F) to the arterial line (5) or the cell separator (7),

[0448] It is characterized in that

[0449] The bypass line section (12') of the bypass line (12) branches off from the plasma line (8A) and is connected to the second single sampling column (4'), and the bypass line section (12") of the bypass line (12") is led out from the single sampling column (4") and connected to the plasma line (8B).

[0450] The waste liquid pipeline (13') is directly led out from the single collection column (4') or is led out from the plasma pipeline (8B) before the confluence of the bypass pipeline section (12') of the bypass pipeline (12) in the flow direction. The waste liquid pipeline (13") is directly led out from the single collection column (4") or is led out from the bypass pipeline section (12') of the bypass pipeline (12) in the flow direction before the confluence of the bypass pipeline section (12') of the bypass pipeline (12). ”), and each liquid container (F) comprises a regeneration line (14), which is led out from each liquid container (F) or their connecting line (11) and flows along the flow direction after the branch of the bypass line section (12') of the bypass line (12) to the plasma line (8A) or the bypass line section (12') of the bypass line (12) or directly connected to the single sampling column (4') or directly connected to the single sampling column (4"), and

[0451] The second single sampling column (4") is connected in parallel with the first single sampling column (4'), and the two single sampling columns (4', 4") cannot be used to remove CRP at the same time, wherein the single sampling device (II) is designed to be tolerant to alkali metal hydroxide solution (preferably sodium hydroxide solution).

[0452] In addition, an embodiment of the single collection device (II) is preferred, wherein the single collection device (II) has at least two connecting lines (11) each for connecting at least one liquid container (F) to the arterial line (5) or the cell separator (7), and wherein each liquid container (F) has a regeneration line (14), which is led out from each liquid container (F) or their connecting lines (11) and is respectively introduced into the plasma line (8A) or into the bypass line section (12') of the bypass line (12) or directly into the single collection column (4') or directly into the single collection column (4").

[0453] According to some embodiments of the present invention, preferably at least one regeneration line (14) leads to the plasma line (8A) or to the bypass line segment (12') of the bypass line (12) or is directly connected to the single sampling column (4') or is directly connected to the single sampling column (4"), starting from the point (P5) where at least one connecting line (11) leads out.

[0454] Preferably, the single collection device (II) has two connecting lines (11', 11') each for connecting one of the liquid containers (F1, F2) to the arterial line (5) or the cell separator (7), and two regeneration lines (14', 14") are led out from the two liquid containers (F1, F2) or the two connecting lines (11', 11") and introduced into the plasma line (8A) or into the bypass line segment (12') of the bypass line (12) or directly into the blood component column (4') or directly into the blood component column (4").

[0455] A particularly preferred embodiment is one in which the regeneration line (14) which is introduced into the plasma line (8A) or into the bypass line section (12') of the bypass line (12) or directly into the single sampling column (4') or directly into the single sampling column (4") and which starts from a point (P5) in at least one connecting line (11) has at least one additional connection ( Fig.10 ). For example, an infusion bag containing an alkali metal hydroxide solution, preferably a sodium hydroxide solution, can be connected to the additional connection piece.

[0456] In an embodiment of the present invention having multiple connecting lines (11', 11", 11"', etc.) and multiple regeneration lines (14', 14", 14"', etc.), it is possible that each connecting line is connected to a corresponding regeneration line, and the regeneration line is connected to the plasma line (8A) or the bypass line segment (12') of the bypass line (12) or directly connected to the single sampling column (4') or directly connected to the single sampling column (4") after point (P2). Here, each regeneration line can be connected to the plasma line (8A) or the bypass line segment (12') of the bypass line (12) or directly connected to the single sampling column (4') or directly enter the single sampling column (4") independently of other regeneration lines. However, it is preferred that all regeneration lines are directly connected to the single sampling column (4', 4"), preferably at point (P2) in the extracorporeal circulation system (2). It can be based on Fig.10 To explain such an exemplary embodiment. Here, the single collection device (II) has a first connecting line (11'), which first leads to the arterial line (5), and secondly branches out a first regeneration line (14') from the connecting line at point (P5'). The single collection device (II) also has a second connecting line (11"), which first directly introduces the cell separator (7), and secondly branches out a second regeneration line (14") from the connecting line at point (P5"). In this embodiment, both regeneration lines enter the extracorporeal circulation system (2) at point (P2).

[0457] Therefore, a single collection device (II) is preferred, wherein the single collection device (II) is each used to connect at least one liquid container (F) to two connecting lines (11', 11") of an arterial line (5) or a cell separator (7), and wherein at least one regeneration line (14), which leads to a plasma line (8A) or is introduced into a bypass line segment (12') of a bypass line (12) or directly leads to a single collection column (4') or is directly introduced into a single collection column (4"), is connected to the connecting line (11') at point (P5') and is connected to the connecting line (11") at point (P5").

[0458] Therefore, an embodiment of the apheresis device is particularly preferred, wherein the apheresis device (II) has two connecting lines (11', 11') each for connecting one of the liquid containers (F1, F2) to the arterial line (5) or the cell separator (7), and wherein at least one regeneration line (14) is introduced into the plasma line (8A) or into the bypass line section (12') of the bypass line (12) or directly into the apheresis column (4') or directly into the apheresis column (4") and is connected to the connecting line (11') at point (P5') and to the connecting line (11') at point (P5"). The pipeline (11") is connected, and the regeneration pipeline (14') is led from the liquid container (F1) or the connecting pipeline (11') led from the liquid container (F1) to the single sampling column (4') or the single sampling column (4") or the plasma pipeline (8A') or the plasma pipeline (8A") and the regeneration pipeline (14") is led from the liquid container (F2) or the connecting pipeline (11") led from the liquid container (F2) to the single sampling column (4') or the single sampling column (4") or the plasma pipeline (8A) or the bypass pipeline section (12') of the bypass pipeline (12) or introduced into the regeneration pipeline (14').

[0459] It is therefore particularly preferred that the apheresis device (II) has a connecting line (11') for connecting to the liquid container (F1) and a connecting line (11") for connecting to the liquid container (F2), the connecting line (11') being connected to the arterial line (5) or the cell separator (7), the connecting line (11") being connected to the arterial line (5) or the cell separator (7) or being connected to the connecting line (11') and thus ultimately also to the arterial line (5) or to the cell separator (7), and the regeneration line The line (14') leads from the liquid container (F1) or from the connecting line (11') to the single sampling column (4') or to the single sampling column (4") or to the plasma line (8A) or to the plasma line (8A") and the regeneration line (14") leads from the liquid container (F2) or from the connecting line (11") to the single sampling column (4') or to the single sampling column (4") or to the plasma line (8A') or to the bypass line section (12') of the bypass line (12) or is introduced into the regeneration line (14').

[0460] Therefore, an embodiment of the apheresis device (II) is particularly preferred, wherein the apheresis device (II) has a connecting line (11') for connecting the liquid container (F1) to the arterial line (5) or the cell separator (7) and a connecting line (11") for connecting the liquid container (F2) to the arterial line (5) or the cell separator (7), and wherein the regeneration line (14') is led out from the liquid container (F1) or the connecting line (11') and introduced into the blood. The plasma pipeline (8A) or the bypass pipeline section (12') of the bypass pipeline (12) or directly introduced into the single sampling column (4') or directly introduced into the single sampling column (4") and the regeneration pipeline (14") are led out from the liquid container (F2) or the connecting pipeline (11") and introduced into the plasma pipeline (8A) or the bypass pipeline section (12') of the bypass pipeline (12) or introduced into the regeneration pipeline (14') or directly introduced into the single sampling column (4') or directly introduced into the single sampling column (4").

[0461] Therefore, the present invention also relates to a single collection device (II) according to the present invention, wherein the plasma line (8A) and the bypass line segment (12') of the bypass line (12) branch from point (P2), and the plasma line (8B) and the bypass line segment (12") of the bypass line (12) and the plasma line (8B) converge at point (P6), and the waste liquid line (13') is led out from point (P4) of the plasma line (8B) and the waste liquid line (13") is led out from point (P8) of the bypass line segment (12") of the bypass line (12), and at least one regeneration line (14) is connected to the extracorporeal circulation system (2) at point (P2).

[0462] A preferred embodiment of the present invention relates to an apheresis device (II) for extracorporeal removal of CRP from blood, comprising:

[0463] Extracorporeal circulation system for blood (2),

[0464] Device (3) for generating and regulating blood flow in an extracorporeal circulation system (2)

[0465] a cell separator (7) for separating blood into plasma and cellular components,

[0466] Two single sampling columns (4', 4") for affinity chromatography removal of CRP from plasma,

[0467] The extracorporeal circulation system (2) comprises an arterial line (5) to a cell separator (7), a plasma line (8A) from the cell separator (7) to a single collection column (4'), and a plasma line (8B) from the single collection column (4') to a point (P1) for plasma depleted of CRP.

[0468] a cell line (9) for separating cell components from the cell separator (7) to the point (P1), and a venous line (6) starting from the point (P1),

[0469] at least one connecting line (11) for connecting at least one liquid container (F) to the arterial line (5) or the cell separator (7),

[0470] Features

[0471] The bypass line section (12') of the bypass line (12) branches off from the plasma line (8A) and is connected to the second single sampling column (4'), and the bypass line section (12") of the bypass line (12") starts from the single sampling column (4") and is connected to the plasma line (8B).

[0472] The waste liquid pipeline (13') is directly led out from the single sampling column (4') or is led out from the plasma pipeline (8B) before the confluence of the bypass pipeline segment (12') of the bypass pipeline (12) in the flow direction, and the waste liquid pipeline (13") is directly led out from the single sampling column (14") or is led out from the bypass pipeline segment (12") of the bypass pipeline (12) before the confluence of the bypass pipeline segment (12') of the bypass pipeline (12) in the flow direction, wherein the plasma pipeline (8A) and the bypass pipeline segment (12') of the bypass pipeline (12) diverge from point (P2), and the plasma pipeline (8B) and the bypass pipeline segment (12") of the bypass pipeline (12) merge at point (P6), and

[0473] at least one regeneration line (14), which leads to the plasma line (8A) or to the bypass line section (12') of the bypass line (12) or directly connects to the single sampling column (4') or directly connects to the single sampling column (4") after the bypass line section (12') of the bypass line (12) in the flow direction, and

[0474] The waste line (13') is led from point (P4) of the plasma line (8B), the waste line (13") is led from point (P8) of the bypass line section (12") of the bypass line (12), and

[0475] At least one regeneration line (14) is connected to the extracorporeal circulation system (2) at point (P2), and

[0476] The second single sampling column (4") is connected in parallel with the first single sampling column (4'), and the two single sampling columns (4', 4") cannot be used to remove CRP at the same time, that is, they can only be used alternately, wherein the single sampling device (II) is designed to be tolerant to alkali metal hydroxide solution (preferably sodium hydroxide solution).

[0477] In order to further reduce the dead volume of the system, it is more preferred that not only the regeneration line (14) is connected to the extracorporeal circulation system at the point (P2) where the plasma line (8A) and the bypass line (12') of the bypass line segment (12) branch off, but also the waste liquid line (13', 13") merges at the same point (P6) where the plasma line (8B) and the bypass line segment (12") of the bypass line (12) branch off. In other words, it is preferred that the point (P6) where the plasma line (8B) and the bypass line segment (12") of the bypass line (12) merge, the point (P8) where the waste line (13") branches off, and the point (P4) where the waste liquid line (13') branches off coincide, that is, when P8 = P4 = P6 (see Figure 8 ).

[0478] Therefore, the present invention also relates to a single collection device (II) according to the present invention, wherein the plasma line (8B) and the bypass line segment (12") of the bypass line (12) merge at point (P6), the waste liquid line (13") is led out from point (P8) of the bypass line segment (12") of the bypass line (12), the waste liquid line (13') is led out from point (P4) of the plasma line (8B), and at least one regeneration line (14) is connected to the extracorporeal circulation system (2) at point (P2), wherein point (P6), point (P4) and point (P8) are the same.

[0479] According to the present invention, an embodiment of the apheresis device (II) for extracorporeal removal of CRP from blood according to the present invention comprises two apheresis columns (4', 4") for affinity chromatography removal of CRP from blood or plasma, the function of which is to bind CRP present in the patient's blood or plasma and to guide it through the apheresis columns (4') or (4").

[0480] method

[0481] Another aspect of the present invention also relates to a method for regenerating an apheresis column (4) for removing CRP by affinity chromatography in an apheresis device (1), which method can be regenerated during continuous operation and is characterized by the following steps:

[0482] (A) starting to transfer the separated plasma from the plasma line (8A) into the bypass line (12), thereby stopping the introduction of the separated plasma from the plasma line (8A) into the apheresis column (4),

[0483] (B) starting to introduce a regeneration solution into the plasma line (8A) or directly into the single sampling column (4) via at least one regeneration line (14), wherein the regeneration solution is an alkali metal hydroxide solution, preferably a sodium hydroxide solution,

[0484] (C) starting to transfer the liquid flow out of the single collection column (4) from the plasma pipeline (8B) to the waste pipeline (13),

[0485] (D) stopping the introduction of the regeneration solution,

[0486] (E) starting to introduce a neutralizing solution,

[0487] (F) stopping the introduction of the neutralizing solution and stopping the transfer of the separated plasma from the plasma line (8A) to the bypass line (12), thereby introducing the separated plasma from the plasma line (8A) into the single collection column (4),

[0488] (G) The waste line (13) is closed and the liquid flow out of the single sampling column (4) is further introduced into the venous line (6).

[0489] Another aspect of the present invention also relates to a method for regenerating an apheresis column (4) for removing CRP by affinity chromatography in an apheresis device (1), the method being capable of being regenerated during continuous operation and comprising the following steps:

[0490] (A) starting to transfer the separated plasma from the plasma line (8A) into the bypass line (12), thereby stopping the introduction of the separated plasma from the plasma line (8A) into the apheresis column (4),

[0491] (B) starting to introduce a regeneration solution into the plasma line (8A) or directly into the single sampling column (4) via at least one regeneration line (14), wherein the regeneration solution is an alkali metal hydroxide solution, preferably a sodium hydroxide solution,

[0492] (C) starting to transfer the liquid flow out of the single collection column (4) from the plasma pipeline (8B) to the waste pipeline (13),

[0493] (D) stopping the introduction of the regeneration solution,

[0494] (E) starting to introduce a neutralizing solution,

[0495] (F) Stop introducing the neutralizing solution.

[0496] Another aspect of the present invention also relates to a method for regenerating an apheresis column (4) for removing CRP by affinity chromatography in an apheresis device (1), the method being capable of being regenerated during continuous operation and comprising the following steps:

[0497] (A) starting to transfer the separated plasma from the plasma line (8A) into the bypass line (12), thereby stopping the introduction of the separated plasma from the plasma line (8A) into the apheresis column (4),

[0498] (B) starting to introduce a regeneration solution into the plasma line (8A) or directly into the single sampling column (4) via at least one regeneration line (14), wherein the regeneration solution is an alkali metal hydroxide solution, preferably a sodium hydroxide solution,

[0499] (C) starting to transfer the liquid flow out of the single collection column (4) from the plasma pipeline (8B) to the waste pipeline (13),

[0500] (D) stopping the introduction of the regeneration solution,

[0501] (E) starting to introduce a neutralizing solution,

[0502] (F) stopping the introduction of the neutralizing solution and stopping the transfer of the separated plasma from the plasma line (8A) to the bypass line (12), thereby introducing the separated plasma from the plasma line (8A) into the single collection column (4),

[0503] (G) Close the waste line (13).

[0504] Another aspect of the present invention also relates to a method for regenerating an apheresis column (4) for removing CRP by affinity chromatography in an apheresis device (1), the method being capable of being regenerated during continuous operation and comprising the following steps:

[0505] (A) starting to transfer the separated plasma from the plasma line (8A) into the bypass line (12), thereby stopping the introduction of the separated plasma from the plasma line (8A) into the apheresis column (4),

[0506] (B) starting to introduce a regeneration solution into the plasma line (8A) or directly into the single sampling column (4) via at least one regeneration line (14), wherein the regeneration solution is an alkali metal hydroxide solution, preferably a sodium hydroxide solution,

[0507] (C) starting to transfer the liquid flow out of the single collection column (4) from the plasma pipeline (8B) to the waste pipeline (13),

[0508] (D) stopping the introduction of the regeneration solution,

[0509] (E) starting to introduce a neutralizing solution,

[0510] (F) The introduction of the neutralizing solution is stopped and the transfer of the separated plasma from the plasma line (8A) to the bypass line (12) is stopped, thereby introducing the separated plasma from the plasma line (8A) into the apheresis column (4).

[0511] The term "stopping the introduction of separated plasma" according to step (A) can be understood as meaning, according to the embodiments of the present invention, using a hose clamp, a control element, a valve and / or a hose pump to prevent the plasma from further flowing into the plasma line (8A) or into the bypass line section (12') of the bypass line (12) or into the single collection column (4') or (4").

[0512] The term "stop introducing the regeneration solution" according to step (D) can be understood to mean, according to an embodiment of the present invention, using a hose clamp, a control element, a valve and / or a peristaltic pump to stop the regeneration solution from flowing further into the plasma line (8A) or the single sampling column (4). Here, it should be understood that in embodiments in which only one regeneration solution is used, the introduction of this regeneration solution is stopped. In embodiments in which multiple regeneration solutions are introduced continuously, this means stopping the introduction of the last regeneration solution used, and thus also stopping the introduction of any regeneration solution.

[0513] The term "closing the waste liquid line (13)" according to step (E) can be understood to mean, depending on the embodiment of the present invention, stopping the further flow of liquid out of the single sampling column (4) using a hose clamp, a control element, a valve and / or a hose pump. Here, it is understood that in embodiments where only one regeneration solution is used, the introduction of this regeneration solution is stopped. In embodiments where multiple regeneration solutions are introduced continuously, it is understood that the introduction of the last used regeneration solution is stopped, and thus also the introduction of any regeneration solution is stopped.

[0514] By "further introducing the liquid flow out of the single collection column (4)" according to step (E), the separated plasma flows into the plasma pipeline (8B) again after flowing through the single collection column (4) and further passes through the vein (6) therefrom. According to an embodiment of the present invention, a hose clamp, a control element, a valve and / or a hose pump can be used to change the flow direction of the liquid flow out of the single collection column (4).

[0515] Furthermore, the present invention relates to a method for regenerating an apheresis column (4) for removing CRP by affinity chromatography in an apheresis device (1) as described herein, wherein the method is capable of being operated continuously by switching from an apheresis mode to a regeneration mode, wherein in the apheresis mode, plasma separated from blood by a cell separator (7) is introduced into the apheresis column (4) via a plasma line (8A), and liquid flowing out of the apheresis column (4) is introduced into a venous line (6) via a plasma line (8B);

[0516] Wherein, the regeneration method is characterized by the following steps:

[0517] (A) starting to transfer the separated plasma from the plasma line (8A) into the bypass line (12), thereby stopping the introduction of the separated plasma from the plasma line (8A) into the apheresis column (4),

[0518] (B) starting to introduce a regeneration solution into the plasma line (8A) or directly into the single sampling column (4) via at least one regeneration line (14), wherein the regeneration solution is an alkali metal hydroxide solution, preferably a sodium hydroxide solution,

[0519] (C) starting to transfer the liquid flow out of the single collection column (4) from the plasma pipeline (8B) to the waste pipeline (13),

[0520] (D) stopping the introduction of the regeneration solution,

[0521] (E) starting to introduce a neutralizing solution,

[0522] (F) stopping the introduction of the neutralizing solution and stopping the transfer of the separated plasma from the plasma line (8A) to the bypass line (12), thereby introducing the separated plasma from the plasma line (8A) into the single collection column (4),

[0523] (E) Close the waste line (13).

[0524] Preferably, step (E) closes the waste line (13) and switches to the single collection mode. It is further preferred that step (E) closes the waste line (13) and further introduces the liquid flow out of the single collection column (4) into the venous line (6) and thereby switches to the single collection mode again.

[0525] For the above two methods, the regeneration solution is preferably an alkali metal hydroxide solution, preferably a sodium hydroxide solution.

[0526] Furthermore, a method is preferred in which step (C) is started after a total volume of X of regeneration solution has been introduced into the plasma line (8A) or directly into the single sampling column (4), wherein X corresponds to at least 75% of the capacity of the device between the point where the regeneration line (14) branches off from the bypass line (12) and joins the extracorporeal circulation system (2) and the point where the waste liquid line (13) is connected from the extracorporeal circulation system (2) in the flow direction. Here, the regeneration solution is, for example, an alkali hydroxide solution, preferably a sodium hydroxide solution.

[0527] Furthermore, a method is preferred in which step (E) is initiated after a volume Y of plasma is introduced into the plasma line (8A) or directly into the single collection column (4), wherein Y corresponds to at least 90% of the capacity of the device between the point where the regeneration line (14) is connected to the extracorporeal circulation system (2) after branching off the bypass line (12) in the flow direction and the point where the waste liquid line (13) is connected from the extracorporeal circulation system (2).

[0528] As used herein, "during continuous operation" means that in order to perform the method for regenerating the single collection column (4) according to the present invention, the operation of the blood extraction and supply and the cell separator does not have to be stopped. In other words, in the regeneration method for the single collection column (4) according to the present invention, the continuously obtained plasma bypasses the single collection column (4) through the bypass line (12) to merge with the cell components and is supplied to the patient. During the regeneration period, the single collection column is therefore disconnected from the blood extraction and supply or the blood circulation. During the period when the plasma diversion flow through the bypass line (12) occurs, the single collection column (4), which usually has a reduced working capacity, is regenerated. Therefore, the patient's circulation is not stressed because the continuously extracted blood is immediately returned to the patient's body.

[0529] Therefore, as used herein, "during continuous operation" does not mean that continuous plasma extraction must be interrupted to implement the method for regenerating the apheresis column (4) of the present invention. In addition, this does not mean that CRP-depletion will occur during the regeneration of the apheresis column.

[0530] Thus, in the above-described methods and the methods generally disclosed herein, preferably the introduction of the regeneration solution comprises the introduction of a single regeneration solution or the continuous introduction of multiple regeneration solutions.

[0531] It is completely clear to the person skilled in the art that an initial flushing step of the adsorber or the entire system must take place before carrying out the method according to the invention. This is associated with the pre-filling of the entire tubing system. For this purpose, in some cases there may be further connections on the system, which allow flushing of the entire system. After the patient has been separated from the tubing system, the adsorber may potentially be stored in order to be used again for further treatment of the same patient.

[0532] In other words, the present invention also relates to a method for regenerating an apheresis column (4) for the removal of CRP by affinity chromatography in an apheresis device (1) according to the present invention, wherein the method enables regeneration during operation and is characterized by the following steps:

[0533] (A) transferring the separated plasma from the plasma line (8A) into the bypass line (12),

[0534] (B) introducing at least one regeneration solution from a liquid container into the plasma line (8A) or directly into the single sampling column (4) via the regeneration line (14), wherein the regeneration solution is an alkali metal hydroxide solution, preferably a sodium hydroxide solution,

[0535] (C) transferring the liquid flow leaving the plasma apheresis column (4) from the plasma pipeline (8B) to the waste pipeline (13),

[0536] (D) stopping the introduction of the regeneration solution,

[0537] (E) starting to introduce a neutralizing solution,

[0538] (F) transferring the separated plasma from the plasma line (8A) to the single sampling column (4) and stopping the introduction of the neutralizing solution,

[0539] (G) Close the waste line (13).

[0540] Preferably, step (G) closes the waste line (13) and further introduces the liquid flow out of the single sampling column (4) into the venous line (6).

[0541] As used herein, the term "turn" refers to a change in the direction of the fluid flow in question. During treatment / processing mode, separated plasma flows into the apheresis column (4) via plasma line (8A). After leaving the apheresis column (4), the depleted plasma flows into the venous line (6) via plasma line (8B).

[0542] By "diverting" the flow direction of the separated plasma according to step (A), the separated plasma no longer flows through the apheresis column (4), but is diverted into the bypass line (12) to bypass it.

[0543] As used herein, the term "introduced" according to step (B) may be understood to mean that, according to an embodiment of the present invention, at least one regeneration solution is supplied (using or operating a hose clamp, a control element, a valve and / or a hose pump) into the plasma line (8A) or into the single sampling column (4).

[0544] By "changing" the flow direction of the liquid flow out of the single sampling column (4) according to step (C), the outflowing liquid no longer flows into the plasma line (8B) but directly flows into the waste liquid line (13). According to the present invention, it is preferred that the waste liquid line (13) branches directly or immediately from the single sampling column (4) or after the single sampling column (4) to minimize the volume of the regeneration solution required to regenerate the single sampling column (4). In the sense of the present invention, the waste liquid line (13) can also branch from the plasma line (8B), so it does not have to branch directly from the single sampling column.

[0545] By the "turn" flow direction of the separated plasma according to step (D), the separated plasma then flows through the single sampling column (4) again and no longer flows into the bypass line (12). In certain embodiments, a pump is provided in the bypass line (12), whereby the plasma present in the bypass line (12) after the turn according to step (D) is pumped into the plasma line (8B) and through the venous line (6). Here, the plasma present in the bypass line is preferably replaced by the NaCl solution from the regeneration line (14). Preferably, a 0.9% NaCl solution is involved here. It is also conceivable that a separate liquid container can be connected to the bypass line (12), through which the NaCl solution is provided for replacement.

[0546] Therefore, a method for regenerating a single sampling column (4) for removing CRP by affinity chromatography in a single sampling device (1) is particularly preferred, characterized by the following steps:

[0547] (A) starting to transfer the separated plasma from the plasma line (8A) into the bypass line (12), thereby stopping the introduction of the separated plasma from the plasma line (8A) into the apheresis column (4),

[0548] (B) starting to introduce a flushing solution into the plasma line (8A) or directly into the single sampling column (4) via at least one regeneration line (14),

[0549] (C) starting to transfer the liquid flow out of the single collection column (4) from the plasma pipeline (8B) to the waste pipeline (13),

[0550] (D) stopping the introduction of the flushing solution and transitioning to introducing the regeneration solution into the plasma line (8A) or directly into the single sampling column (4) via at least one regeneration line (14), wherein the regeneration solution is an alkali metal hydroxide solution (preferably a sodium hydroxide solution),

[0551] (E) stopping the introduction of the regeneration solution and transitioning to introducing the flushing solution into the plasma line (8A) or directly into the single sampling column (4) via at least one regeneration line (14),

[0552] (F) stopping the introduction of the flushing solution and stopping the transfer of the separated plasma from the plasma line (8A) into the bypass line (12), and thereby introducing the separated plasma from the plasma line (8A) into the apheresis column (4);

[0553] (G) Close the waste line (13).

[0554] Preferably, step (G) closes the waste line (13) and further introduces the liquid flow out of the single sampling column (4) into the venous line (6).

[0555] Alternatively, a method for regenerating an apheresis column (4) for removing CRP by affinity chromatography in an apheresis device (1) is particularly preferred, wherein the method is characterized by the following steps:

[0556] (A) starting to transfer the separated plasma from the plasma line (8A) into the bypass line (12), thereby stopping the introduction of the separated plasma from the plasma line (8A) into the apheresis column (4),

[0557] (B) starting to introduce the flushing solution into the plasma line (8A) or directly into the single sampling column (4) via at least one regeneration line (14),

[0558] (C) stopping the introduction of the flushing solution and transitioning to introducing the regeneration solution into the plasma line (8A) or directly into the single sampling column (4) via at least one regeneration line (14), wherein the regeneration solution is an alkali metal hydroxide solution (preferably a sodium hydroxide solution),

[0559] (D) starting to transfer the liquid flow out of the single collection column (4) from the plasma pipeline (8B) to the waste pipeline (13),

[0560] (E) stopping the introduction of the regeneration solution and transitioning to introducing the flushing solution into the plasma line (8A) or directly into the single sampling column (4) via at least one regeneration line (14),

[0561] (F) closing the waste liquid pipeline (13);

[0562] (G) Stopping the introduction of flushing solution and stopping the transfer of separated plasma from the plasma line (8A) into the bypass line (12) and thereby introducing the separated plasma from the plasma line (8A) into the apheresis column (4).

[0563] Preferably, step (F) closes the waste line (13) and further introduces the liquid flow out of the single sampling column (4) into the venous line (6).

[0564] In both methods, in addition to the regeneration solution, a flushing solution is used. The flushing solution is preferably physiologically compatible and is mainly used to displace plasma from the plasma line (8A) from point P2, from the single sampling column (4) and from the plasma line (8B) to point P4. The flushing solution has little or no effect on the regeneration of the single sampling column (4). The regeneration solution is introduced to regenerate the single sampling column (4) only when the plasma has been substantially to completely displaced from the part of the single sampling device (1) flushed with the regeneration solution. After regeneration, the flushing solution is first fed again into the part of the single sampling device (1) that has been flushed with the regeneration solution (i.e. in the flow direction from point P2 through the single sampling column (4) to point P4) until the regeneration solution is completely discharged through the waste liquid line (13). Only then is the bypass line (12) closed and the plasma is again introduced through the single sampling column (4). In both methods, steps (C) and (D) can be interchanged, that is, they can be carried out in any order, can be carried out simultaneously, or can be combined into one step. However, it is preferred that step (D) is performed before step (C).

[0565] In the method, the flushing solution is preferably a physiological NaCl solution or a PBS solution, and the regeneration solution is an alkali metal hydroxide solution (preferably a sodium hydroxide solution).

[0566] Therefore, a method for regenerating a single sampling column (4) for removing CRP by affinity chromatography in a single sampling device (1) is particularly preferred, characterized by the following steps:

[0567] (A) starting to transfer the separated plasma from the plasma line (8A) into the bypass line (12), thereby stopping the introduction of the separated plasma from the plasma line (8A) into the apheresis column (4),

[0568] (B) starting to introduce the flushing solution into the plasma line (8A) or directly into the single sampling column (4) via at least one regeneration line (14),

[0569] (C) starting to transfer the liquid flow out of the single collection column (4) from the plasma pipeline (8B) to the waste pipeline (13),

[0570] (D) stopping the introduction of the flushing solution and transitioning to introducing the regeneration solution into the plasma line (8A) or directly into the single sampling column (4) via at least one regeneration line (14), wherein the regeneration solution is an alkali metal hydroxide solution (preferably a sodium hydroxide solution),

[0571] (E) stopping the introduction of the regeneration solution and transitioning to introducing the neutralization solution into the plasma line (8A) or directly into the single sampling column (4) via at least one regeneration line (14),

[0572] (F) stopping the introduction of the neutralization solution and transitioning to the introduction of the flushing solution into the plasma line (8A) via at least one regeneration line (14) or directly into the single sampling column (4),

[0573] (G) stopping the introduction of the flushing solution and stopping the transfer of the separated plasma from the plasma line (8A) into the bypass line (12) and thereby introducing the separated plasma from the plasma line (8A) into the apheresis column (4);

[0574] (H) Close the waste line (13).

[0575] Preferably, step (H) closes the waste line (13) and further introduces the liquid flow out of the single sampling column (4) into the venous line (6).

[0576] Alternatively, a method for regenerating an apheresis column (4) for removing CRP by affinity chromatography in an apheresis device (1) is particularly preferred, wherein the method is characterized by the following steps:

[0577] (A) starting to transfer the separated plasma from the plasma line (8A) into the bypass line (12), thereby stopping the introduction of the separated plasma from the plasma line (8A) into the apheresis column (4),

[0578] (B) starting to introduce the flushing solution into the plasma line (8A) or directly into the single sampling column (4) via at least one regeneration line (14),

[0579] (C) stopping the introduction of the flushing solution and transitioning to introducing the regeneration solution into the plasma line (8A) or directly into the single sampling column (4) via at least one regeneration line (14), wherein the regeneration solution is an alkali metal hydroxide solution (preferably a sodium hydroxide solution),

[0580] (D) starting to transfer the liquid flow out of the single collection column (4) from the plasma pipeline (8B) to the waste pipeline (13),

[0581] (E) stopping the introduction of the regeneration solution and transitioning to introducing the neutralization solution into the plasma line (8A) or directly into the single sampling column (4) via at least one regeneration line (14),

[0582] (F) stopping the introduction of the neutralization solution and transitioning to the introduction of the flushing solution into the plasma line (8A) via at least one regeneration line (14) or directly into the single sampling column (4),

[0583] (G) closing the waste liquid pipeline (13);

[0584] (H) The introduction of the flushing solution is stopped and the transfer of the separated plasma from the plasma line (8A) into the bypass line (12) and thereby the introduction of the separated plasma from the plasma line (8A) into the apheresis column (4) is stopped.

[0585] Preferably, step (G) closes the waste line (13) and further introduces the liquid flow out of the single sampling column (4) into the venous line (6).

[0586] In the above two methods, steps (C) and (D) are interchangeable, i.e., they can be performed in any order, simultaneously, or combined in one step. However, it is preferred that step (D) is performed before step (C).

[0587] A preferred embodiment of the method according to the invention is used to perform the method more efficiently without loss of plasma. By simultaneously diverting the separated plasma and introducing the flushing solution in parallel to the apheresis column (4), no or no significant loss of plasma occurs. In addition, the preferred embodiment has the advantage that mixing of the regeneration solution with the plasma is completely avoided. This ensures that no regeneration solution enters the patient, and on the other hand, the patient's plasma is not lost.

[0588] This is ensured by the sequence of steps (B) to (E). Dilution of the plasma, if any, is only carried out by the flushing solution. On the other hand, mixing of the plasma with the regeneration solution is completely avoided.

[0589] The volume of the washing solution according to step (B) preferably corresponds to 3 to 4 times the matrix volume of the apheresis column (4).

[0590] Minimally, the volume of flushing solution according to step (B) corresponds to the capacity of the plasma line (8A) from point P2 to the single collection column (4) plus the matrix volume of the single collection column (4) and plus the capacity of the plasma line (8B) from the single collection column (4) to point P4.

[0591] The volume of the regeneration solution in step (C) is preferably 2 to 100 times the volume of the matrix of the single sampling column (4).

[0592] The volume of the washing solution according to step (E) preferably corresponds to 2 to 4 times the matrix volume of the apheresis column (4).

[0593] The volume of the flushing solution according to at least step (E) corresponds to the capacity of the plasma line (8A) from point P2 to the single collection column (4) plus the matrix volume of the single collection column (4) and plus the capacity of the plasma line (8B) from the single collection column (4) to point P4.

[0594] According to this even more preferred embodiment, dilution of the plasma is largely avoided and mixing with the regeneration solution is completely prevented. The user does not encounter too many complications when using the apheresis device (1). In an alternative embodiment, the method steps can therefore also be performed manually without this being too complicated for the user.

[0595] As used herein, the "matrix volume of the single sampling column" refers to the volume of the solid phase in the column, which in turn includes the matrix substrate material and the compound with specific binding properties of CRP bound thereto. In contrast, the "dead volume of the single sampling column" is the space in the column that can be used for the mobile phase (e.g., plasma). The "dead volume of the single sampling column" is the difference between the volume enclosed by the shell of the single sampling column and the volume occupied by the expanded matrix (i.e., the "matrix volume of the single sampling column").

[0596] Another aspect of the present invention relates to a method for regenerating an apheresis column (4') for removing CRP by affinity chromatography during the continuous operation of a second apheresis column (4") in an apheresis device (II), comprising the following steps:

[0597] (A) starting from the plasma flow through the single sampling column (4"), starting to introduce the separated plasma into the single sampling column (4') through the plasma pipeline (8A), and stopping the introduction of the separated plasma into the single sampling column (4") through the bypass pipeline section (12') of the bypass pipeline (12),

[0598] (B) starting to introduce a regeneration solution into the bypass line section (12') of the bypass line (12) or directly into the single sampling column (4") via at least one regeneration line (14), wherein at least one regeneration solution is an alkali metal hydroxide solution (preferably a sodium hydroxide solution),

[0599] (C) starting to divert the liquid flow out of the blood apheresis column (4") from the bypass line section (12") of the bypass line (12) into the waste liquid line (13"),

[0600] (D) starting to introduce the neutralization solution into the bypass line section (12') of the bypass line (12) via at least one regeneration line (14) or directly into the single sampling column (4"),

[0601] (E) stopping the introduction of the neutralizing solution,

[0602] (F) starting to introduce the separated plasma into the single sampling column (4") through the bypass line section (12') of the bypass line (12) and stopping introducing the separated plasma into the single sampling column (4') through the plasma line (8A),

[0603] (G) Close the waste line (13") and begin diverting the flow of liquid out of the apheresis column (4') from the plasma line (8B) to the waste line (13').

[0604] Preferably, step (A) starts with the plasma flowing through the single sampling column (4"), begins to introduce the separated plasma into the single sampling column (4') via the plasma line (8A) and introduces the CRP-depleted plasma into the venous line (6), and thereby stops introducing the separated plasma into the single sampling column (4") via the bypass portion (12') of the bypass line (12).

[0605] Preferably, step (F) begins introducing separated plasma into the apheresis column (4") via the bypass line segment (12') of the bypass line (12) and introducing CRP-depleted plasma into the venous line (6), thereby stopping the introduction of separated plasma into the apheresis column (4') via the plasma line (8A).

[0606] As used herein, "during continuous operation" means that in order to perform the method for regenerating a single collection column (4') or regenerating a single collection column (4") of the present invention, it is not necessary to stop the blood collection blood supply and the operation of the cell separator. Therefore, the patient's circulation will not be stressed because the continuously extracted blood will be immediately returned to the patient.

[0607] In other words, in one embodiment, the present invention relates to a method for use in a single collection device (II), comprising the following steps:

[0608] (A) starting from the plasma flow through the single sampling column (4"), the separated plasma is introduced into the single sampling column (4') via the plasma pipeline (8A) and the introduction of the separated plasma into the single sampling column (4") via the bypass pipeline section (12') of the bypass pipeline (12) is stopped,

[0609] (B) introducing at least one regeneration solution into the bypass line section (12') of the bypass line (12) or directly into the single sampling column (4") via at least one regeneration line (14), wherein at least one regeneration solution is an alkali metal hydroxide solution (preferably a sodium hydroxide solution),

[0610] (C) diverting the liquid flow out of the blood apheresis column (4") from the bypass line section (12") of the bypass line (12) into the waste liquid line (13"),

[0611] (D) starting to introduce the neutralization solution into the bypass line section (12') of the bypass line (12) via at least one regeneration line (14) or directly into the single sampling column (4"),

[0612] (E) stopping the introduction of the neutralizing solution,

[0613] (F) Introducing the separated plasma into the single sampling column (4") via the bypass line section (12') of the bypass line (12), and stopping the introduction of the separated plasma into the single sampling column (4') via the plasma line (8A)

[0614] (G) Close the waste line (13") and divert the liquid flow out of the apheresis column (4') from the plasma line (8B) to the waste line (13').

[0615] Preferably, step (A) starts from the plasma flow through the single sampling column (4"), introduces the separated plasma into the single sampling column (4') via the plasma line (8A), and introduces the CRP-depleted plasma into the venous line (6) and thereby stops the introduction of the separated plasma into the single sampling column (4") via the bypass line section (12') of the bypass line (12).

[0616] Preferably, step (F) introduces the separated plasma into the apheresis column (4") via the bypass line section (12') of the bypass line (12), and introduces the CRP-depleted plasma into the venous line (6) and thereby stops the introduction of the plasma into the apheresis column (4') via the plasma line (8A).

[0617] Furthermore, the present invention relates to a method for regenerating two apheresis columns (4', 4") for the removal of CRP by affinity chromatography in an apheresis device (II), wherein the method enables regeneration during continuous operation, characterized by the following steps:

[0618] (A) starting from the plasma flow through the single sampling column (4"), starting to introduce the separated plasma into the single sampling column (4') via the plasma line (8A), and stopping introducing the separated plasma into the single sampling column (4") via the bypass line segment (12') of the bypass line (12),

[0619] (B) starting to introduce the flushing solution into the bypass line section (12') of the bypass line (12) via at least one regeneration line (14) or directly into the single sampling column (4"),

[0620] (C) starting to introduce the liquid flow out of the single sampling column (4") from the bypass line section (12") of the bypass line (12) into the waste liquid line (13");

[0621] (D) stopping the introduction of the flushing solution and transitioning to introducing the regeneration solution into the bypass line section (12') of the bypass line (12) or directly into the single sampling column (4") via at least one regeneration line (14), wherein the regeneration solution is an alkali metal hydroxide solution (preferably a sodium hydroxide solution),

[0622] (E) stopping the introduction of the regeneration solution and transitioning to introducing the flushing solution into the bypass pipeline section (12') of the bypass pipeline (12) or directly into the single sampling column (4") via at least one regeneration pipeline (14),

[0623] (F) starting to introduce a flushing solution into the plasma line (8A) through the single sampling column (4'), and thereby introducing the separated plasma into the single sampling column (4") via the bypass line section (12') of the bypass line (12),

[0624] (G) Close the waste line (13”),

[0625] (H) starting to transfer the liquid flow out of the single collection column (4') from the plasma pipeline (8B) to the waste pipeline (13'),

[0626] (I) Stop introducing the flushing solution and transition to introducing the regeneration solution into the plasma line (8A) or directly into the single sampling column (4') via at least one regeneration line (14).

[0627] (J) Stopping the introduction of the regeneration solution and transitioning to introducing the flushing solution into the plasma line (8A) or directly into the apheresis column (4') via at least one regeneration line (14).

[0628] Preferably, step (A) starts with the plasma flow through the single sampling column (4"), begins to introduce the separated plasma into the single sampling column (4') via the plasma line (8A) and introduces the CRP-depleted plasma into the venous line (6), thereby stopping the introduction of the separated plasma into the single sampling column (4") via the bypass line segment (12') of the bypass line (12).

[0629] Preferably, step (G) closes the waste line (13") and diverts the liquid flow out of the single sampling column (4") into the venous line (6).

[0630] In both of the above methods, a flushing solution is used in addition to the regeneration solution. The flushing solution is preferably physiologically acceptable and is mainly used to replace the plasma in the plasma line (8A) or in the bypass line segment (12') of the bypass line (12) starting from point P2, in the single sampling column (4') or the single sampling column (4"), and in the bypass line segment (12") of the bypass line (12) to point P3 and in the plasma line (8B) to point P4. The flushing solution is less or not used to regenerate the single sampling column (4') or the single sampling column (4"). Therefore, the use of the flushing solution can minimize or even completely prevent plasma loss. The regeneration solution is introduced to regenerate the single sampling column (4') or the single sampling column (4") only when the plasma has been basically to completely replaced from the portion of the single sampling device (II) to be flushed with the regeneration solution. After regeneration is completed, the flushing solution is first reintroduced into the portion of the single sampling device (II) that has been flushed with the regeneration solution (i.e., along the flow direction from point P2 through the single sampling column (4") to point P8) or through the single sampling column (4') to point P4), until the regeneration solution is completely discharged through the waste liquid pipeline (13', 13").

[0631] In these present methods, the flushing solution is preferably physiological saline or PBS solution (phosphate buffered saline) or a combination of physiological saline and PBS solution sequentially or simultaneously, and the regeneration solution is an alkali metal hydroxide solution, preferably a sodium hydroxide solution.

[0632] Furthermore, the present invention relates to a method for regenerating two apheresis columns (4', 4") for the removal of CRP by affinity chromatography in an apheresis device (II), wherein the method enables regeneration during continuous operation, characterized by the following steps:

[0633] (A) starting from the plasma flow through the single sampling column (4"), starting to introduce the separated plasma into the single sampling column (4') via the plasma line (8A), and stopping introducing the separated plasma into the single sampling column (4") via the bypass line segment (12') of the bypass line (12),

[0634] (B) starting to introduce the flushing solution into the bypass line section (12') of the bypass line (12) via at least one regeneration line (14) or directly into the single sampling column (4"),

[0635] (C) starting to introduce the liquid flow out of the single sampling column (4") from the bypass line section (12") of the bypass line (12) into the waste liquid line (13");

[0636] (D) stopping the introduction of the flushing solution and transitioning to introducing the regeneration solution into the bypass line section (12') of the bypass line (12) or directly into the single sampling column (4") via at least one regeneration line (14), wherein the regeneration solution is an alkali metal hydroxide solution (preferably a sodium hydroxide solution),

[0637] (E) stopping the introduction of the regeneration solution and transitioning to introducing the neutralization solution into the bypass pipeline section (12') of the bypass pipeline (12) via at least one regeneration pipeline (14) or directly into the single sampling column (4"),

[0638] (F) stopping the introduction of the neutralization solution and transitioning to introducing the neutralization solution into the bypass pipeline section (12') of the bypass pipeline (12) via at least one regeneration pipeline (14) or directly into the single sampling column (4"),

[0639] (G) starting to introduce a flushing solution into the plasma line (8A) through the single sampling column (4'), and thereby introducing the separated plasma into the single sampling column (4") via the bypass line section (12') of the bypass line (12),

[0640] (H) Close the waste line (13”),

[0641] (I) starting to transfer the liquid flow out of the single collection column (4') from the plasma pipeline (8B) to the waste liquid pipeline (13'),

[0642] (I) Stop introducing the flushing solution and transition to introducing the regeneration solution into the plasma line (8A) or directly into the single sampling column (4') via at least one regeneration line (14).

[0643] (J) Stopping the introduction of the regeneration solution and transitioning to introducing the flushing solution into the plasma line (8A) or directly into the apheresis column (4') via at least one regeneration line (14).

[0644] Preferably, step (A) starts with the plasma flow through the single sampling column (4"), begins to introduce the separated plasma into the single sampling column (4') via the plasma line (8A) and introduces the CRP-depleted plasma into the venous line (6), thereby stopping the introduction of the separated plasma into the single sampling column (4") via the bypass line segment (12') of the bypass line (12).

[0645] Preferably, step (H) closes the waste line (13") and diverts the liquid flow out of the single sampling column (4") into the venous line (6).

[0646] In the above two methods, steps (C) and (D) can be interchanged, that is, they can be performed in any order, can be performed simultaneously, or can be combined into one step. However, it is preferred to perform step (D) before step (C).

[0647] A preferred embodiment of the method according to the invention is provided for carrying out the method efficiently without loss of plasma. By simultaneously diverting the separated plasma and introducing the flushing solution in parallel to the single collection column (4"), no or no significant loss of plasma is achieved. Furthermore, an advantage of the preferred embodiment is that mixing of regeneration solution and plasma is completely avoided. This ensures that no regeneration solution enters the patient and, on the other hand, the patient's plasma is not lost.

[0648] This is ensured by the sequence of steps (B) to (E). Dilution of the plasma, if any, is only carried out by the flushing solution. On the other hand, mixing of the plasma with the regeneration solution is completely avoided.

[0649] The volume of the flushing solution according to step (B) preferably corresponds to 3 to 4 times the volume of the matrix of the single sampling column (4"). At the very least, the volume of the flushing solution according to step (B) corresponds to the capacity of the bypass line section (12') of the bypass line (12) from point P2 to the single sampling column (4") plus the volume of the matrix of the single sampling column (4") plus the capacity of the bypass line section (12') of the bypass line (12) from the single sampling column (4") to point (P3).

[0650] The volume of the flushing solution according to step (F) preferably corresponds to 3 to 4 times the volume of the matrix of the apheresis column (4'). Minimally, the volume of the flushing solution according to step (B) corresponds to the volume of the plasma line (8A') from point (P2) to the apheresis column (4') plus the volume of the matrix of the apheresis column (4') plus the volume of the plasma line (8B) from the apheresis column to point (P4).

[0651] The volume of the regeneration solution according to step (D) is preferably 2 to 100 times the volume of the matrix of the single sampling column (4").

[0652] The volume of the regeneration solution according to step (I) preferably corresponds to 2 to 100 times the matrix volume of the single sampling column (4').

[0653] The volume of the washing solution according to step (E) preferably corresponds to 2 to 4 times the matrix volume of the apheresis column (4").

[0654] The volume of the flushing solution according to step (E) corresponds at least to the volume of the bypass line section (12') of the bypass line (12) from point P2 to the single sampling column (4") plus the matrix volume of the single sampling column (4") plus the volume of the bypass line section (12') of the bypass line (12) from the single sampling column (4') to point P3.

[0655] The volume of the washing solution according to step (J) preferably corresponds to 2 to 4 times the matrix volume of the apheresis column (4').

[0656] The volume of the flushing solution according to step (E) corresponds at least to the capacity of the plasma line (8A) from point P2 to the single sampling column (4') plus the matrix volume of the single sampling column (4'), and plus the capacity of the plasma line (8B) from the single sampling column (4') to point P4.

[0657] In a preferred embodiment, the flushing solution is a saline solution or a physiological saline solution or a PBS solution (phosphate buffered saline) or a combination of a saline solution and a PBS solution sequentially or simultaneously, and the regeneration solution is an alkali metal hydroxide solution (preferably a sodium hydroxide solution).

[0658] Furthermore, a preferred embodiment relates to a method for regenerating an apheresis column (4) for removing CRP by affinity chromatography in an apheresis device (1), wherein the method is characterized by the following steps:

[0659] (A) starting to transfer the separated plasma from the plasma line (8A) into the bypass line (12), thereby stopping the introduction of the separated plasma from the plasma line (8A) into the apheresis column (4),

[0660] (B) starting to introduce saline solution into the plasma line (8A) via at least one regeneration line (14) or directly into the single sampling column (4),

[0661] (C) starting to transfer the liquid flow out of the single collection column (4) from the plasma pipeline (8B) to the waste pipeline (13),

[0662] (D) stopping the introduction of the saline solution and transitioning to introducing the alkali metal hydroxide solution (preferably sodium hydroxide solution) into the plasma pipeline (8A) via at least one regeneration pipeline (14) or directly into the single sampling column (4),

[0663] (E) stopping the introduction of alkali metal hydroxide solution (preferably sodium hydroxide solution) and transitioning to introducing saline solution into the plasma pipeline (8A) or directly into the single sampling column (4) via at least one regeneration pipeline (14),

[0664] (F) stopping the introduction of saline solution and stopping the transfer of separated plasma from the plasma line (8A) into the bypass line (12) and thereby introducing the separated plasma from the plasma line (8A) into the apheresis column (4);

[0665] (G) Close the waste line (13).

[0666] Preferably, step (G) closes the waste line (13) and further introduces the liquid flow out of the single sampling column (4) into the venous line (6).

[0667] Alternatively, a preferred embodiment relates to a method for regenerating an apheresis column (4) for removing CRP by affinity chromatography in an apheresis device (1), wherein the method is characterized by the following steps:

[0668] (A) starting to transfer the separated plasma from the plasma line (8A) into the bypass line (12), thereby stopping the introduction of the separated plasma from the plasma line (8A) into the apheresis column (4),

[0669] (B) starting to introduce saline solution into the plasma line (8A) via at least one regeneration line (14) or directly into the single sampling column (4),

[0670] (C) stopping the introduction of the saline solution and transitioning to introducing the alkali metal hydroxide solution (preferably sodium hydroxide solution) into the plasma pipeline (8A) via at least one regeneration pipeline (14) or directly into the single sampling column (4),

[0671] (D) starting to transfer the liquid flow out of the single collection column (4) from the plasma pipeline (8B) to the waste pipeline (13),

[0672] (E) stopping the introduction of alkali metal hydroxide solution (preferably sodium hydroxide solution) and transitioning to introducing saline solution into the plasma pipeline (8A) or directly into the single sampling column (4) via at least one regeneration pipeline (14),

[0673] (F) closing the waste liquid pipeline (13);

[0674] (G) The introduction of the saline solution is stopped and the transfer of the separated plasma from the plasma line (8A) to the bypass line (12) is stopped, thereby introducing the separated plasma from the plasma line (8A) into the apheresis column (4).

[0675] Preferably, step (F) closes the waste line (13) and further introduces the liquid flow out of the single sampling column (4) into the venous line (6).

[0676] Furthermore, a preferred embodiment relates to a method for regenerating an apheresis column (4) for removing CRP by affinity chromatography in an apheresis device (1), wherein the method is characterized by the following steps:

[0677] (A) starting to transfer the separated plasma from the plasma line (8A) into the bypass line (12), thereby stopping the introduction of the separated plasma from the plasma line (8A) into the apheresis column (4),

[0678] (B) starting to introduce saline solution into the plasma line (8A) via at least one regeneration line (14) or directly into the single sampling column (4),

[0679] (C) starting to transfer the liquid flow out of the single collection column (4) from the plasma pipeline (8B) to the waste pipeline (13),

[0680] (D) stopping the introduction of the saline solution and transitioning to introducing an alkali metal hydroxide solution (preferably a sodium hydroxide solution) into the plasma line (8A) via at least one regeneration line (14) or directly into the single sampling column (4),

[0681] (E) stopping the introduction of the alkali metal hydroxide solution (preferably sodium hydroxide solution) and transitioning to introducing the citrate solution into the plasma pipeline (8A) or directly into the single sampling column (4) via at least one regeneration pipeline (14),

[0682] (F) stopping the introduction of the citrate solution and transitioning to introducing the saline solution into the plasma line (8A) via at least one regeneration line (14) or directly into the apheresis column (4),

[0683] (G) stopping the introduction of saline solution and stopping the transfer of separated plasma from the plasma line (8A) to the bypass line (12), thereby introducing the separated plasma from the plasma line (8A) into the apheresis column (4);

[0684] (H) Close the waste line (13).

[0685] Preferably, step (H) closes the waste line (13) and further introduces the liquid flow out of the single sampling column (4) into the venous line (6).

[0686] Alternatively, a preferred embodiment relates to a method for regenerating an apheresis column (4) for removing CRP by affinity chromatography in an apheresis device (1), wherein the method is characterized by the following steps:

[0687] (A) starting to transfer the separated plasma from the plasma line (8A) into the bypass line (12), thereby stopping the introduction of the separated plasma from the plasma line (8A) into the apheresis column (4),

[0688] (B) starting to introduce saline solution into the plasma line (8A) via at least one regeneration line (14) or directly into the single sampling column (4),

[0689] (C) stopping the introduction of the saline solution and transitioning to introducing the alkali metal hydroxide solution (preferably sodium hydroxide solution) into the plasma pipeline (8A) via at least one regeneration pipeline (14) or directly into the single sampling column (4),

[0690] (D) starting to transfer the liquid flow out of the single collection column (4) from the plasma pipeline (8B) to the waste pipeline (13),

[0691] (E) stopping the introduction of the alkali metal hydroxide solution (preferably sodium hydroxide solution) and transitioning to introducing the citrate solution into the plasma pipeline (8A) or directly into the single sampling column (4) via at least one regeneration pipeline (14),

[0692] (F) stopping the introduction of the citrate solution and transitioning to introducing the saline solution into the plasma line (8A) via at least one regeneration line (14) or directly into the apheresis column (4),

[0693] (G) closing the waste liquid pipeline (13);

[0694] (H) The introduction of saline is stopped and the transfer of the separated plasma from the plasma line (8A) to the bypass line (12) is stopped, thereby introducing the separated plasma from the plasma line (8A) into the apheresis column (4).

[0695] Preferably, step (G) closes the waste line (13) and directs the liquid flow out of the single sampling column (4) into the venous line (6).

[0696] In the above four methods, steps (C) and (D) can be interchanged, that is, they can be performed in any order, can be performed simultaneously, or can be combined into one step.

[0697] Preferably, the regeneration method according to the invention is carried out in such a way that the plasma is first displaced from the apheresis column (4) with a flushing solution (e.g. a saline solution or a physiological saline solution) and returned to the patient until almost only saline solution is returned. Only then is the saline introduced into the waste line (13), and a regeneration solution, e.g. an alkali metal hydroxide solution (preferably a sodium hydroxide solution), is introduced into the plasma line (8A) after the bypass line (12) in the flow direction, which displaces the saline, regenerates the apheresis column (4), is completely introduced into the waste line (13) and is discarded. After the apheresis column (4) has been regenerated with several apheresis column volumes of the regeneration solution, a flushing solution, e.g. a saline solution or a physiological saline solution, is introduced again until the regeneration solution is completely displaced from the apheresis device (1) and discarded. Only then are the waste line (13) closed, the flushing solution returned to the patient, the bypass line (12) closed and the plasma reintroduced into the single collection column (4) via the plasma line (8A) simultaneously or directly sequentially, wherein the order of the steps can be interchanged.

[0698] Another preferred embodiment relates to a method for regenerating an apheresis column (4) for the removal of CRP by affinity chromatography in an apheresis device (1), wherein the method is characterized by the following steps:

[0699] (A) starting to transfer the separated plasma from the plasma line (8A) into the bypass line (12), thereby stopping the introduction of the separated plasma from the plasma line (8A) into the apheresis column (4),

[0700] (B) starting to introduce saline solution into the plasma line (8A) via at least one regeneration line (14) or directly into the single sampling column (4),

[0701] (C) starting to transfer the liquid flow out of the single collection column (4) from the plasma pipeline (8B) to the waste pipeline (13),

[0702] (D) stopping the introduction of the saline solution and transitioning to introducing the alkali metal hydroxide solution (preferably sodium hydroxide solution) into the plasma pipeline (8A) via at least one regeneration pipeline (14) or directly into the single sampling column (4),

[0703] (E1) stopping the introduction of alkali metal hydroxide solution (preferably sodium hydroxide solution) and transitioning to introducing saline solution into the plasma pipeline (8A) or directly into the single sampling column (4) via at least one regeneration pipeline (14),

[0704] (E2) stopping the introduction of saline solution and transitioning to introducing PBS solution into the plasma line (8A) via at least one regeneration line (14) or directly into the single sampling column (4),

[0705] (E3) stopping the introduction of PBS solution and transitioning to introducing saline solution into the plasma line (8A) via at least one regeneration line (14) or directly into the single sampling column (4),

[0706] (F) stopping the introduction of saline solution and stopping the transfer of separated plasma from the plasma line (8A) to the bypass line (12), thereby introducing the separated plasma from the plasma line (8A) into the apheresis column (4);

[0707] (G) Close the waste line (13).

[0708] Preferably, step (G) closes the waste line (13) and further introduces the liquid flow out of the single sampling column (4) into the venous line (6).

[0709] Another preferred embodiment relates to a method for regenerating an apheresis column (4) for the removal of CRP by affinity chromatography in an apheresis device (1), wherein the method is characterized by the following steps:

[0710] (A) starting to transfer the separated plasma from the plasma line (8A) into the bypass line (12), thereby stopping the introduction of the separated plasma from the plasma line (8A) into the apheresis column (4),

[0711] (B) starting to introduce saline solution into the plasma line (8A) via at least one regeneration line (14) or directly into the single sampling column (4),

[0712] (C) stopping the introduction of the saline solution and transitioning to introducing the alkali metal hydroxide solution (preferably sodium hydroxide solution) into the plasma pipeline (8A) via at least one regeneration pipeline (14) or directly into the single sampling column (4),

[0713] (D) starting to transfer the liquid flow out of the single collection column (4) from the plasma pipeline (8B) to the waste pipeline (13),

[0714] (E1) stopping the introduction of alkali metal hydroxide solution (preferably sodium hydroxide solution) and transitioning to introducing saline solution into the plasma pipeline (8A) or directly into the single sampling column (4) via at least one regeneration pipeline (14),

[0715] (E2) stopping the introduction of saline solution and transitioning to introducing PBS solution into the plasma line (8A) via at least one regeneration line (14) or directly into the single sampling column (4),

[0716] (E3) stopping the introduction of PBS solution and transitioning to introducing saline solution into the plasma line (8A) via at least one regeneration line (14) or directly into the single sampling column (4),

[0717] (F) closing the waste liquid pipeline (13);

[0718] (G) The introduction of the saline solution and the transfer of the separated plasma from the plasma line (8A) to the bypass line (12) are stopped, thereby introducing the separated plasma from the plasma line (8A) into the apheresis column (4).

[0719] Preferably, step (F) closes the waste line (13) and further introduces the liquid flow out of the single sampling column (4) into the venous line (6).

[0720] Another preferred embodiment relates to a method for regenerating an apheresis column (4) for removing CRP by affinity chromatography in an apheresis device (1), wherein the method is characterized by the following steps:

[0721] (A) starting to transfer the separated plasma from the plasma line (8A) into the bypass line (12), thereby stopping the introduction of the separated plasma from the plasma line (8A) into the apheresis column (4),

[0722] (B) starting to introduce saline solution into the plasma line (8A) via at least one regeneration line (14) or directly into the single sampling column (4),

[0723] (C) starting to transfer the liquid flow out of the single collection column (4) from the plasma pipeline (8B) to the waste pipeline (13),

[0724] (D) stopping the introduction of the saline solution and transitioning to introducing an alkali metal hydroxide solution (preferably a sodium hydroxide solution) into the plasma line (8A) via at least one regeneration line (14) or directly into the single sampling column (4),

[0725] (E1) stopping the introduction of alkali metal hydroxide solution (preferably sodium hydroxide solution) and transitioning to introducing citrate solution into the plasma pipeline (8A) or directly into the single sampling column (4) via at least one regeneration pipeline (14),

[0726] (E2) stopping the introduction of the citrate solution and transitioning to introducing the saline solution into the plasma line (8A) via at least one regeneration line (14) or directly into the single sampling column (4),

[0727] (E3) stopping the introduction of saline solution and transitioning to introducing PBS solution into the plasma line (8A) via at least one regeneration line (14) or directly into the single sampling column (4),

[0728] (E4) stopping the introduction of PBS solution and transitioning to introducing saline solution into the plasma line (8A) via at least one regeneration line (14) or directly into the single sampling column (4),

[0729] (F) stopping the introduction of saline and stopping the transfer of separated plasma from the plasma line (8A) to the bypass line (12), thereby introducing the separated plasma from the plasma line (8A) into the apheresis column (4);

[0730] (G) Close the waste line (13).

[0731] Preferably, step (G) closes the waste line (13) and further introduces the liquid flow out of the single sampling column (4) into the venous line (6).

[0732] Another preferred embodiment relates to a method for regenerating an apheresis column (4) for the removal of CRP by affinity chromatography in an apheresis device (1), wherein the method is characterized by the following steps:

[0733] (A) starting to transfer the separated plasma from the plasma line (8A) into the bypass line (12), thereby stopping the introduction of the separated plasma from the plasma line (8A) into the apheresis column (4),

[0734] (B) starting to introduce saline solution into the plasma line (8A) via at least one regeneration line (14) or directly into the single sampling column (4),

[0735] (C) stopping the introduction of the saline solution and transitioning to introducing the alkali metal hydroxide solution (preferably sodium hydroxide solution) into the plasma pipeline (8A) via at least one regeneration pipeline (14) or directly into the single sampling column (4),

[0736] (D) starting to transfer the liquid flow out of the single collection column (4) from the plasma pipeline (8B) to the waste pipeline (13),

[0737] (E1) stopping the introduction of alkali metal hydroxide solution (preferably sodium hydroxide solution) and transitioning to introducing citrate solution into the plasma pipeline (8A) via at least one regeneration pipeline (14) or directly into the single sampling column (4),

[0738] (E2) stopping the introduction of the citrate solution and transitioning to introducing the saline solution into the plasma line (8A) via at least one regeneration line (14) or directly into the single sampling column (4),

[0739] (E3) stopping the introduction of saline solution and transitioning to introducing PBS solution into the plasma line (8A) via at least one regeneration line (14) or directly into the single sampling column (4),

[0740] (E4) stopping the introduction of PBS solution and transitioning to introducing saline solution into the plasma line (8A) via at least one regeneration line (14) or directly into the single sampling column (4),

[0741] (F) closing the waste liquid pipeline (13);

[0742] (G) The introduction of the saline solution and the transfer of the separated plasma from the plasma line (8A) to the bypass line (12) are stopped, thereby introducing the separated plasma from the plasma line (8A) into the apheresis column (4).

[0743] Preferably, step (F) closes the waste line (13) and further introduces the liquid flow out of the single sampling column (4) into the venous line (6).

[0744] In the above four methods, steps (C) and (D) can be interchanged, that is, they can be performed in any order, can be performed simultaneously, or can be combined in one step.

[0745] The present invention therefore relates to a method for regenerating two apheresis columns (4', 4") for the removal of CRP by affinity chromatography in an apheresis device (II), wherein the method enables regeneration during continuous operation and is characterized by the following steps:

[0746] (A) starting from the plasma flow through the single sampling column (4"), starting to introduce the separated plasma into the single sampling column (4') via the plasma line (8A), and stopping introducing the separated plasma into the single sampling column (4") via the bypass line segment (12') of the bypass line (12),

[0747] (B) starting to introduce the brine solution into the bypass line section (12') of the bypass line (12) via at least one regeneration line (14) or directly into the single sampling column (4"),

[0748] (C) starting to introduce the liquid flow out of the single sampling column (4") from the bypass line section (12") of the bypass line (12) into the waste liquid line (13");

[0749] (D) stopping the introduction of the brine solution and transitioning to introducing the alkali metal hydroxide solution (preferably sodium hydroxide solution) into the bypass line section (12') of the bypass line (12) via at least one regeneration line (14) or directly connecting to the single sampling column (4"),

[0750] (E) stopping the introduction of alkali metal hydroxide solution (preferably sodium hydroxide solution) and transitioning to introducing the salt solution into the bypass pipeline section (12') of the bypass pipeline (12) via at least one regeneration pipeline (14) or directly connecting it to the single sampling column (4"),

[0751] (F) starting to introduce saline solution into the plasma pipeline (8A) via the single sampling column (4'), thereby introducing the separated plasma into the single sampling column (4") via the bypass pipeline section (12') of the bypass pipeline (12),

[0752] (G) Close the waste line (13”),

[0753] (H) starting to transfer the liquid flow out of the single collection column (4') from the plasma pipeline (8B) to the waste pipeline (13'),

[0754] (I) stopping the introduction of the flushing solution and transitioning to introducing the alkali metal hydroxide solution (preferably sodium hydroxide solution) into the plasma pipeline (8A) or directly into the single sampling column (4') via at least one regeneration pipeline (14),

[0755] (J) Stopping the introduction of alkali metal hydroxide solution (preferably sodium hydroxide solution) and transitioning to introducing saline solution into the plasma line (8A) or directly into the single sampling column (4') via at least one regeneration line (14).

[0756] Preferably, step (A) starts with the plasma flow through the single sampling column (4"), begins to introduce the separated plasma into the single sampling column (4') via the plasma line (8A) and introduces the CRP-depleted plasma into the venous line (6), thereby stopping the introduction of the separated plasma into the single sampling column (4") via the bypass line segment (12') of the bypass line (12).

[0757] Preferably, step (G) closes the waste line (13") and diverts the liquid flow out of the single sampling column (4") into the venous line (6).

[0758] The present invention therefore relates to a method for regenerating two apheresis columns (4', 4") for the removal of CRP by affinity chromatography in an apheresis device (II), wherein the method enables regeneration during continuous operation and is characterized by the following steps:

[0759] (A) starting from the plasma flow through the single sampling column (4"), starting to introduce the separated plasma into the single sampling column (4') via the plasma line (8A), and stopping introducing the separated plasma into the single sampling column (4") via the bypass line segment (12') of the bypass line (12),

[0760] (B) starting to introduce the brine solution into the bypass line section (12') of the bypass line (12) via at least one regeneration line (14) or directly into the single sampling column (4"),

[0761] (C) starting to introduce the liquid flow out of the single sampling column (4") from the bypass line section (12") of the bypass line (12) into the waste liquid line (13");

[0762] (D) stopping the introduction of the brine solution and transitioning to introducing the alkali metal hydroxide solution (preferably sodium hydroxide solution) into the bypass line section (12') of the bypass line (12) via at least one regeneration line (14) or directly connecting to the single sampling column (4"),

[0763] (E) stopping the introduction of alkali metal hydroxide solution (preferably sodium hydroxide solution) and transitioning to introducing the salt solution into the bypass pipeline section (12') of the bypass pipeline (12) via at least one regeneration pipeline (14) or directly connecting it to the single sampling column (4"),

[0764] (F) starting to introduce saline solution into the plasma pipeline (8A) via the single sampling column (4'), thereby introducing the separated plasma into the single sampling column (4") via the bypass pipeline section (12') of the bypass pipeline (12),

[0765] (G) Close the waste line (13”),

[0766] (H) starting to transfer the liquid flow out of the single collection column (4') from the plasma pipeline (8B) to the waste pipeline (13'),

[0767] (I) stopping the introduction of the flushing solution and transitioning to introducing the alkali metal hydroxide solution (preferably sodium hydroxide solution) into the plasma pipeline (8A) or directly into the single sampling column (4') via at least one regeneration pipeline (13),

[0768] (J) stopping the introduction of alkali metal hydroxide solution (preferably sodium hydroxide solution) and transitioning to introducing citrate solution into the plasma pipeline (8A) or directly into the single sampling column (4') via at least one regeneration pipeline (14),

[0769] (K) Stopping the introduction of citrate solution and transitioning to introducing saline solution into the plasma line (8A) via at least one regeneration line (14) or directly into the apheresis column (4').

[0770] Preferably, step (A) starts with the plasma flow through the single sampling column (4"), begins to introduce the separated plasma into the single sampling column (4') via the plasma line (8A) and introduces the CRP-depleted plasma into the venous line (6), thereby stopping the introduction of the separated plasma into the single sampling column (4") via the bypass line segment (12') of the bypass line (12).

[0771] Preferably, step (G) closes the waste line (13") and diverts the liquid flow out of the single sampling column (4") into the venous line (6).

[0772] Preferably, the regeneration method according to the present invention is carried out in such a way that first the plasma is displaced from the apheresis column (4") with a flushing solution (e.g. a saline solution or a physiological saline solution) until almost only the saline solution passes through. The saline solution is then preferably introduced into the waste line (13"), and a regeneration solution, e.g. an alkali metal hydroxide solution (preferably a sodium hydroxide solution) is introduced into the bypass line section (12') of the bypass line (12) at point P2 along the flow direction, which displaces the saline solution, regenerates the apheresis column (4"), is completely introduced into the waste line (13") and discarded. After the apheresis column (4") is regenerated with several apheresis column volumes of the regeneration solution, a flushing solution, e.g. a saline solution or a physiological saline solution, is introduced again until the regeneration solution is completely displaced from the apheresis device (II) and discarded. Only then is the plasma line (8A) closed simultaneously or directly in sequence, the flushing solution is returned to the patient, and the plasma is reintroduced into the apheresis column (4") through the bypass line section (12') of the bypass line (12).

[0773] Another preferred embodiment relates to a method for regenerating two apheresis columns (4', 4") for the removal of CRP by affinity chromatography in an apheresis device (II), wherein the method is capable of regeneration during continuous operation and is characterized by the following steps:

[0774] (A) starting from the plasma flow through the single sampling column (4"), starting to introduce the separated plasma into the single sampling column (4') via the plasma line (8A), and stopping introducing the separated plasma into the single sampling column (4") via the bypass line segment (12') of the bypass line (12),

[0775] (B) starting to introduce the brine solution into the bypass line section (12') of the bypass line (12) via at least one regeneration line (14) or directly into the single sampling column (4"),

[0776] (C) starting to introduce the liquid flow out of the single sampling column (4") from the bypass line section (12') of the bypass line (12) into the waste liquid line (13");

[0777] (D) stopping the introduction of the brine solution and transitioning to introducing the alkali metal hydroxide solution (preferably sodium hydroxide solution) into the bypass line section (12') of the bypass line (12) via at least one regeneration line (14) or directly into the single sampling column (4"),

[0778] (E1) stopping the introduction of alkali metal hydroxide solution (preferably sodium hydroxide solution) and transitioning to introducing the salt solution into the bypass pipeline section (12') of the bypass pipeline (12) via at least one regeneration pipeline (14) or directly connecting to the single sampling column (4"),

[0779] (E2) stopping the introduction of the saline solution and transitioning to introducing the PBS solution into the bypass line section (12') of the bypass line (12) via at least one regeneration line (14) or directly into the single sampling column (4"),

[0780] (E3) stopping the introduction of PBS solution and transitioning to introducing the saline solution into the bypass line section (12') of the bypass line (12) via at least one regeneration line (14) or directly into the single sampling column (4"),

[0781] (F) starting to introduce saline solution into the plasma pipeline (8A) via the single sampling column (4'), thereby introducing the separated plasma into the single sampling column (4") via the bypass pipeline section (12') of the bypass pipeline (12),

[0782] (G) Close the waste line (13”),

[0783] (H) starting to transfer the liquid flow out of the single collection column (4') from the plasma pipeline (8B) to the waste pipeline (13'),

[0784] (I) stopping the introduction of the flushing solution and transitioning to introducing the alkali metal hydroxide solution (preferably sodium hydroxide solution) into the plasma pipeline (8A) or directly into the single sampling column (4') via at least one regeneration pipeline (14),

[0785] (J) Stopping the introduction of alkali metal hydroxide solution (preferably sodium hydroxide solution) and transitioning to introducing saline solution into the plasma line (8A) or directly into the single sampling column (4') via at least one regeneration line (14).

[0786] Preferably, step (A) starts with the plasma flow through the single sampling column (4"), begins to introduce the separated plasma into the single sampling column (4') via the plasma line (8A) and introduces the CRP-depleted plasma into the venous line (6), thereby stopping the introduction of the separated plasma into the single sampling column (4") via the bypass line segment (12') of the bypass line (12).

[0787] Preferably, step (G) closes the waste line (13") and diverts the liquid flow out of the single sampling column (4") into the venous line (6).

[0788] Another preferred embodiment relates to a method for regenerating two apheresis columns (4', 4") for the removal of CRP by affinity chromatography in an apheresis device (II), wherein the method is capable of regeneration during continuous operation and is characterized by the following steps:

[0789] (A) starting from the plasma flow through the single sampling column (4"), starting to introduce the separated plasma into the single sampling column (4') via the plasma line (8A), and stopping introducing the separated plasma into the single sampling column (4") via the bypass line segment (12') of the bypass line (12),

[0790] (B) starting to introduce the brine solution into the bypass line section (12') of the bypass line (12) via at least one regeneration line (14) or directly into the single sampling column (4"),

[0791] (C) starting to introduce the liquid flow out of the single sampling column (4") from the bypass line section (12') of the bypass line (12) into the waste liquid line (13");

[0792] (D) stopping the introduction of the brine solution and transitioning to introducing the alkali metal hydroxide solution (preferably sodium hydroxide solution) into the bypass line section (12') of the bypass line (12) via at least one regeneration line (14) or directly into the single sampling column (4"),

[0793] (E1) stopping the introduction of alkali metal hydroxide solution (preferably sodium hydroxide solution) and transitioning to introducing the salt solution into the bypass line section (12') of the bypass line (12) or directly into the single sampling column (4") via at least one regeneration line (14),

[0794] (E2) stopping the introduction of the saline solution and transitioning to introducing the PBS solution into the bypass line section (12') of the bypass line (12) via at least one regeneration line (14) or directly into the single sampling column (4"),

[0795] (E3) stopping the introduction of PBS solution and transitioning to introducing the saline solution into the bypass line section (12') of the bypass line (12) via at least one regeneration line (14) or directly into the single sampling column (4"),

[0796] (F) starting to introduce saline solution into the plasma pipeline (8A) via the single sampling column (4'), thereby introducing the separated plasma into the single sampling column (4") via the bypass pipeline section (12') of the bypass pipeline (12),

[0797] (G) Close the waste line (13”),

[0798] (H) starting to transfer the liquid flow out of the single collection column (4') from the plasma pipeline (8B) to the waste pipeline (13'),

[0799] (I) stopping the introduction of the flushing solution and transitioning to introducing the alkali metal hydroxide solution (preferably sodium hydroxide solution) into the plasma pipeline (8A) or directly into the single sampling column (4') via at least one regeneration pipeline (14),

[0800] (J) stopping the introduction of alkali metal hydroxide solution (preferably sodium hydroxide solution) and transitioning to introducing citrate solution into the plasma pipeline (8A) or directly into the single sampling column (4') via at least one regeneration pipeline (14),

[0801] (K) Stopping the introduction of citrate solution and transitioning to introducing saline solution into the plasma line (8A) via at least one regeneration line (14) or directly into the apheresis column (4').

[0802] Preferably, step (A) starts with the plasma flow through the single sampling column (4"), begins to introduce the separated plasma into the single sampling column (4') via the plasma line (8A) and introduces the CRP-depleted plasma into the venous line (6), thereby stopping the introduction of the separated plasma into the single sampling column (4") via the bypass line segment (12') of the bypass line (12).

[0803] Preferably, step (G) closes the waste line (13") and diverts the liquid flow out of the single sampling column (4") into the venous line (6).

[0804] In all of the above-mentioned methods, the part of the apheresis apparatus into which the alkali metal hydroxide solution is introduced is resistant to the alkali metal hydroxide solution used, in particular the sodium hydroxide solution used. DETAILED DESCRIPTION

[0805] Application examples:

[0806] As used herein, the term "substrate volume" (also abbreviated MV) refers to the volume of substrate contained within the adsorber.

[0807] As used herein, the term "adsorber capacity" (also abbreviated AV) refers to the capacity of the adsorber housing.

[0808] Example 1: Single sampling with bypass line and single sampling column

[0809] Prepare:

[0810] According to the attached Figure 7 , suitable tubing is placed in the apheresis device (1) for the extracorporeal removal of CRP from the patient's blood, wherein the plasma centrifuge serves as a cell separator (7). A 5 L bag with 0.9% NaCl solution and a 500 ml bag with ACD-A solution (acid-citrate-dextrose solution) and / or a 2000 ml bag with 0.08 M sodium hydroxide solution and / or a 2000 ml bag with glycine / HCl and / or a 2000 ml bag of PBS solution are connected to the connection line or the regeneration line. Two 3 L waste bags are connected to the waste line (13) (e.g., via a three-way valve).

[0811] The arterial (5) and venous (6) lines are connected to each other via adapters. Likewise, the plasma lines (8A and 8B) before and after the adsorber are connected with adapters (without adsorber in between), thereby forming a closed system.

[0812] The entire system is filled with NaCl solution by pre-rinsing with 1 L of 0.9% NaCl solution (200 ml / min); the air present is displaced into the first waste bag. The shaken CRP-adsorber (MV 20 ml, AV 30 ml) is then inserted into the plasma lines (8A and 8B) instead of the adapter. The adsorber is pre-rinsed with 1 L of NaCl solution (100 ml / min). NaCl is also introduced into the first waste bag.

[0813] As a final step of preparation, the plasma centrifuge is pre-filled with 0.9% NaCl solution and 1:15 diluted ACD-A solution. The required capacity consists of the capacity of the piping system in the plasma centrifuge (7), the connecting line to the plasma centrifuge (11) and the plasma line between the plasma centrifuge and P2. The replaced NaCl is introduced into the first waste bag via P4 / P6.

[0814] Single sampling:

[0815] 1. After preparation, switch to the second waste bag. Connect the patient to the arterial (5) and venous (6) lines. At the beginning of apheresis, the blood is introduced into the centrifuge (60-80 ml / min). Throughout the treatment, ACD-A is mixed into the blood via the connecting line (11) at a ratio of 1:15 (1 ml ACD-A per 15 ml blood).

[0816] The replaced NaCl is directed to the second waste bag via P2, bypass line (12) and P4 / P6. Plasma separation begins, and after the corresponding volume of the pipeline from the plasma centrifuge to the node P4 / 6, the system is switched so that the plasma flows into the venous line (6) and back into the patient. After 3 minutes, a constant plasma of about 30 ml / min is reached and the first cycle can begin.

[0817] 2. Close the bypass line (12) and allow the plasma to pass through the adsorber (loading). The NaCl present in the plasma lines (8A and 8B) and the adsorber is introduced into the second waste bag via P4 / P6, and its volume is up to the volume consisting of the plasma lines (8A and 8B) plus the volume of AV. The adsorber is then loaded with 50-100 MV (1000 to 2000 ml) of plasma. After that, regeneration begins.

[0818] 3. Regeneration

[0819] Variant A - Glycine / HCl and PBS solutions

[0820] For this purpose, the plasma is returned to the patient via the bypass line (12).

[0821] The adsorber is now flushed with 0.9% NaCl (30 ml / min) via the regeneration line (14) and the plasma line (8A and 8B). The volume required for this is calculated from the capacity of the AV and the plasma line (8A and 8B). The plasma in the plasma line (8A and 8B) and the adsorber is also returned to the patient, up to a volume consisting of the AV and 75% of the capacity of the plasma line (8A and 8B). P4 / P6 is then switched to introduce the solution into the second waste bag.

[0822] In the next step, it is regenerated (100 ml / min) with 3 MV (60 ml) of 0.9% NaCl and then with 4 MV (80 ml) of glycine / HCl. It is then neutralized with 4 MV (80 ml) of PBS. Afterwards, it is rinsed with 0.9% NaCl (100 ml / min). The volume required for this is calculated from the capacities of the AV, the regeneration line (14) and the plasma lines (8A and 8B).

[0823] Then you can do step 2 (loading) again, followed by step 3.

[0824] Variant B - Sodium hydroxide solution and PBS solution

[0825] For this purpose, the plasma is returned to the patient via the bypass line (12).

[0826] The adsorber is now flushed with 0.9% NaCl (30 ml / min-40 ml / min) via the regeneration line (14) and the plasma line (8A and 8B). The volume required for this is calculated from the capacity of the AV and the plasma line (8A and 8B). The plasma present in the plasma line (8A and 8B) and the adsorber is also returned to the patient in a volume consisting of the AV and 75% of the capacity of the plasma line (8A and 8B). P4 / P6 is then switched to introduce the solution into the second waste bag.

[0827] In the next step, 3 MV (60 ml) of 0.9% NaCl was used for pre-washing, and then 5 MV of 0.08 M NaOH (pH 12.6; flow rate 80 ml / min) was used for regeneration. Then 6 MV of PBS solution was used for neutralization. Then PBS was replaced with 4 MV of 0.9% NaCl (flow rate 80 ml / min).

[0828] Step 2 (loading) can then be performed again, followed by step 3. If necessary, the bag containing the sodium hydroxide solution or PBS solution, respectively, must be replaced.

[0829] Variant C - Sodium hydroxide solu...

Claims

1. Use of an alkali metal hydroxide solution for regenerating a single sampling column, wherein the single sampling column is a single sampling column used for selectively removing C-reactive protein (CRP) from blood or plasma by affinity chromatography, The single sampling column comprises a matrix for removing CRP by affinity chromatography, wherein the matrix comprises a matrix substrate material bound to a ligand, and the ligand has a property of specifically binding to CRP. wherein the ligand bound to the matrix substrate material is selected from phosphorylcholine or phosphorylethanolamine, or wherein the single sampling column comprises a matrix substrate material functionalized with at least one ω-phosphonooxyalkylammonium group and / or at least one ω-ammonium alkoxy-hydroxyphosphoryloxy group, CRP binds to the matrix in the apheresis column with a higher affinity than other substances present in the blood.

2. The use of the alkali metal hydroxide solution according to claim 1, wherein the alkali metal hydroxide solution is a lithium hydroxide solution, a sodium hydroxide solution, a potassium hydroxide solution or a mixture of the above solutions.

3. Use of the alkali metal hydroxide solution according to claim 1, wherein the concentration of the alkali metal hydroxide in the alkali metal hydroxide solution is in the range of 0.01 mol / l to 1 mol / l.

4. Use of the alkali metal hydroxide solution according to claim 3, wherein the concentration of the alkali metal hydroxide in the alkali metal hydroxide solution is in the range of 0.04 mol / l to 0.4 mol / l.

5. Use of the alkali metal hydroxide solution according to claim 1, wherein the pH value of the alkali metal hydroxide solution is in the range of 12 to 14.

6. Use of the alkali metal hydroxide solution according to claim 5, wherein the pH value of the alkali metal hydroxide solution is in the range of 12-13.

7.

7. Use of an alkali metal hydroxide solution according to claim 1, wherein the regeneration is performed during an apheresis treatment for extracorporeal removal of CRP from blood or plasma, wherein the apheresis column is not connected to an extracorporeal circulation system during the regeneration.

8. Use of an alkali metal hydroxide solution according to claim 1, wherein the patient's plasma has been passed through the apheresis column before regeneration.

9. The use of an alkali metal hydroxide solution according to claim 1, wherein the CRP bound to the single sampling column is eluted.

10. The use of an alkali metal hydroxide solution according to claim 1, wherein the bound CRP is eluted and the apheresis column is simultaneously restored to a therapeutically usable state.

11. The use of the alkali metal hydroxide solution according to claim 1, wherein the single sampling column is saturated with CRP.

12. Use of an alkali metal hydroxide solution according to claim 1, wherein the at least one ω-phosphonooxyalkylammonium group corresponds to a group of the following general formula (I): in n is selected from 2 and 3; R 1 and R 2 Independently selected from: –H, –CH3, –C2H5, –C3H7, –C4H9, –C5H 11 , –C6H 13 , or R 1 and R 2 Together with the nitrogen atom to which they are attached, they form a heterocyclic ring selected from: One or more hydrogen atoms are unsubstituted or substituted by fluorine atoms.

13. Use of an alkali metal hydroxide solution according to claim 1, wherein the at least one ω-ammonium alkoxy-hydroxyphosphoryloxy group corresponds to a group of the following general formula (II): in n is selected from 2 and 3; R 1 , R 2 and R 3 Independently selected from: –H, –CH3, –C2H5, –C3H7, –C4H9, –C5H 11 , –C6H 13 , or R 1 and R 2 Together with the nitrogen atom to which they are attached, they form a heterocyclic ring selected from: and R 3 Selected from: –H, –CH3, –C2H5, –C3H7, –C4H9, –C5H 11 , –C6H 13 ; One or more hydrogen atoms are unsubstituted or substituted by fluorine atoms.

14. The use of the alkali metal hydroxide solution according to claim 13, wherein R 3 is -H.

15. The use of an alkali metal hydroxide solution according to claim 1, wherein the single sampling column comprises agarose.

16. Use of an alkali metal hydroxide solution according to any one of claims 1 to 15, wherein the single sampling column contains DNA and / or RNA.

17. A method for regenerating an apheresis column for selectively removing C-reactive protein (CRP) from blood or plasma by affinity chromatography, comprising the following steps: (I) introducing an alkali metal hydroxide solution into a single sampling column for regeneration of the single sampling column, The single sampling column comprises a matrix for removing CRP by affinity chromatography, wherein the matrix comprises a matrix substrate material bound to a ligand, and the ligand has a property of specifically binding to CRP. wherein the ligand bound to the matrix substrate material is selected from phosphorylcholine or phosphorylethanolamine, or wherein the single sampling column comprises a matrix substrate material functionalized with at least one ω-phosphonooxyalkylammonium group and / or at least one ω-ammonium alkoxy-hydroxyphosphoryloxy group, CRP binds to the matrix in the apheresis column with a higher affinity than other substances present in the blood.

18. The method for regenerating an apheresis column for selectively removing CRP from blood or plasma by affinity chromatography according to claim 17, comprising the following steps: (I) introducing an alkali metal hydroxide solution into a single sampling column for regeneration of the single sampling column; and (II) Introducing a neutralizing solution.

19. The method for regenerating an apheresis column for selectively removing CRP from blood or plasma by affinity chromatography according to claim 18, comprising the following steps: (I') introducing a flushing solution into the single sampling column; (I) introducing an alkali metal hydroxide solution into a single sampling column for regeneration of the single sampling column; (II') stopping the introduction of the alkali metal hydroxide solution after step (I); and (II) Introducing a neutralizing solution.

20. The method of claim 19, wherein the apheresis column comprises plasma.

21. The method according to any one of claims 17 to 20, wherein the alkali metal hydroxide solution is a sodium hydroxide solution.

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