Using an alkalihydroxide solution for the regeneration of an apheresis column

1 . A method of using an alkali hydroxide solution for regeneration of an apheresis column, wherein the apheresis column is an apheresis column for the affinity chromatographic selective removal of human C-reactive protein (CRP) from blood or blood plasma,

wherein the apheresis column contains a matrix for affinity chromatographic removal of human CRP, the matrix comprising a matrix substrate material to which ligands are bound which have the property of specifically binding CRP, the ligands bound to the matrix substrate material being selected from phosphocholine or phosphoethanolamine, or wherein the matrix comprises a matrix substrate material functionalized with at least one ω-phosphonooxyalkylammonium group and/or at least one ω-ammoniumalkoxy-hydroxyphosphoryloxy group; and

wherein CRP binds to the matrix in the apheresis column with higher affinity than other substances present in the blood,

the method being performed after blood or blood plasma of a patient has been passed through the apheresis column to remove CRP during a therapeutic apheresis treatment of the patient, wherein the apheresis column contains bound CRP, the method comprising:

(a) introducing a rinsing solution into the apheresis column to remove blood or blood plasma from the column, wherein the rinsing solution is a sodium chloride solution or a phosphate-buffered saline solution; and

(b) introducing the alkali hydroxide solution into the apheresis column for regeneration of the apheresis column,

wherein bound CRP is eluted and the apheresis column is simultaneously restored to a therapeutically usable state, and

wherein the concentration of the alkali hydroxide in the alkali hydroxide solution is in a range of 0.04 mol/l to 0.4 mol/l.

2 . The method of using an alkali hydroxide solution according to claim 1 , wherein the alkali hydroxide solution is a lithium hydroxide solution, a sodium hydroxide solution, a potassium hydroxide solution, or a mixture of the aforementioned solutions.

3 . The method of using an alkali hydroxide solution according to claim 1 , wherein the concentration of the alkali hydroxide in the alkali hydroxide solution is in a range of 0.08 mol/l to 0.1 mol/l.

4 . The method of using an alkali hydroxide solution according to claim 1 , wherein the concentration of the alkali hydroxide in the alkali hydroxide solution is 0.084 mol/l.

5 . The method of using an alkali hydroxide solution according to claim 1 , wherein the alkali hydroxide solution has a pH in a range of 12 to 14.

6 . The method of using an alkali hydroxide solution according to claim 1 , wherein the alkali hydroxide solution has a pH in a range of 12-13.7.

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

8 . The method of using an alkali hydroxide solution according to claim 1 , wherein the apheresis column is saturated with CRP.

9 . The method of using an alkali hydroxide solution according to claim 1 , wherein the apheresis column comprises a matrix substrate material functionalized with at least one ω-phosphonooxyalkylammonium group and/or at least one ω-ammoniumalkoxy-hydroxyphosphoryloxy group.

10 . The method of using an alkali hydroxide solution according to claim 9 , wherein the at least one ω-phosphonooxyalkylammonium group corresponds to a group represented by the following general formula (I)

wherein

n is selected from 2 and 3;

R 1 and R 2 are independently of each other selected from: —H, —CH 3 , —C 2 H 5 ,

—C 3 H 7 , —C 4 H 9 , —C 5 H 11 , —C 6 H 13 , or R 1 and R 2 together with the nitrogen atom to which they are bound can form a heterocycle selected from:

wherein one or more hydrogen atom(s) may be replaced by (a) fluorine atom(s).

11 . The method of using an alkali hydroxide solution according to claim 9 , wherein the at least one ω-ammoniumalkoxy-hydroxyphosphoryloxy group corresponds to a group of the following general formula (II)

wherein

n is selected from 2 and 3;

R 1 , R 2 and R 3 are independently of each other selected from: —H, —CH 3 , —C 2 H 5 , —C 3 H 7 , —C 4 H 9 , —C 5 H 11 , —C 6 H 13 ,

or R 1 and R 2 together with the nitrogen atom to which they are bound can form a heterocycle selected from:

and

R 3 is selected from: —H, —CH 3 , —C 2 H 5 , —C 3 H 7 , —C 4 H 9 , —C 5 H 11 , —C 6 H 13 , and preferably-H;

wherein one or more hydrogen atom(s) may be replaced by (a) fluorine atom(s).

12 . The method of using an alkali hydroxide solution according to claim 1 , wherein the apheresis column contains agarose.

13 . The method of using an alkali hydroxide solution according to claim 1 , wherein the apheresis column contains DNA and/or RNA.

14 . A method for regeneration of an apheresis column for affinity chromatographic selective removal of C-reactive protein (CRP) from blood or blood plasma, the method being performed after blood or blood plasma of a patient has been passed through the apheresis column to remove CRP during a therapeutic apheresis treatment of the patient, wherein the apheresis column contains bound CRP, the method comprising the steps:

(I′) introducing a rinsing solution into an apheresis column preferably containing blood plasma; wherein the rinsing solution is a sodium chloride solution or a phosphate buffered saline solution;

(I) introducing an alkali hydroxide solution, preferably a sodium hydroxide solution, into an apheresis column for regeneration of the apheresis column,

wherein the apheresis column contains a matrix for affinity chromatographic removal of CRP, the matrix comprising a matrix substrate material to which ligands are bound which have the property of specifically binding CRP, the ligands bound to the matrix substrate material being selected from phosphocholine or phosphoethanolamine, or

wherein the matrix comprises a matrix substrate material functionalized with at least one ω-phosphonooxyalkylammonium group and/or at least one ω-ammoniumalkoxy-hydroxyphosphoryloxy group; and

wherein CRP binds to the matrix in the apheresis column with higher affinity than other substances present in the blood,

wherein the concentration of the alkali hydroxide in the alkali hydroxide solution is in a range of 0.04 mol/l to 0.4 mol/l.

15 . The method for regeneration of an apheresis column for affinity chromatographic selective removal of CRP from blood or blood plasma according to claim 14 comprising the steps:

(I′) introducing a rinsing solution into an apheresis column preferably containing blood plasma; wherein the rinsing solution is a sodium chloride solution or a phosphate buffered saline solution;

(I) introducing an alkali hydroxide solution, preferably a sodium hydroxide solution, into an apheresis column for regeneration of the apheresis column; and

(II) introducing a neutralization solution.

16 . The method for regeneration of an apheresis column for affinity chromatographic selective removal of CRP from blood or blood plasma according to claim 14 comprising the steps:

(I′) introducing a rinsing solution into an apheresis column preferably containing blood plasma;

(I) introducing an alkali hydroxide solution, preferably a sodium hydroxide solution, into the apheresis column for regeneration of the apheresis column;

(II″) stopping the introduction of the alkali hydroxide solution, preferably the sodium hydroxide solution, after step (I); and

(II) introducing a neutralization solution.

17 . A method for regeneration of an apheresis column for affinity chromatographic selective removal of C-reactive protein (CRP) from blood or blood plasma in an apheresis device comprising:

an extracorporeal circulation system for blood,

means for generation and regulation of a flow of the blood in the extracorporeal circulation system,

a cell separator for separation of the blood into blood plasma and cellular components,

at least one apheresis column for affinity chromatographic removal of CRP from the blood,

wherein the extracorporeal circulation system comprises an arterial line to the cell separator, a plasma line from the cell separator to the apheresis column, a plasma line for CRP-depleted blood plasma from the apheresis column to a point, a cell line for separated cellular components from the cell separator to the point, and a venous line starting from the point,

at least one connection line for connection of at least one liquid container to the arterial line or the cell separator,

a bypass line branching off from the plasma line and running into the plasma line,

a waste line branching off directly from the apheresis column or from the plasma line in the direction of flow before the junction of the bypass line, and

at least one regeneration line leading in the direction of flow at or after the branch of the bypass line to the plasma line or running directly into the apheresis column, the apheresis device being configured to be resistant to an alkali hydroxide solution, the method enabling regeneration during operation and being performed after blood or blood plasma of a patient has been passed through the apheresis column to remove CRP during a therapeutic apheresis treatment of the patient, wherein the apheresis column contains bound CRP, the method being characterized by the following steps:

(A) starting the redirection of the separated plasma from the plasma line into the bypass line, thereby stopping the introduction of the separated plasma from the plasma line into the apheresis column,

(B) starting the introduction of a rinsing solution via the at least one regeneration line into the plasma line or directly into the apheresis column, wherein the rinsing solution is a sodium chloride solution or a phosphate buffered saline solution;

(C) starting the redirection of the liquid flow exiting the apheresis column from the plasma line into the waste line,

(D) stopping the introduction of the rinsing solution and transition to the introduction of a regeneration solution via the at least one regeneration line into the plasma line or directly into the apheresis column, wherein the regeneration solution is an alkali hydroxide solution,

(E) starting the redirection of the liquid flow exiting the apheresis column from the plasma line to the waste line,

(F) stopping the introduction of regeneration solution,

(G) starting the introduction of neutralization solution,

(H) stopping the introduction of neutralization solution and stopping the redirection of the separated plasma from the plasma line into the bypass line, thereby introduction of the separated plasma from the plasma line into the apheresis column,

(I) closing the waste line,

wherein the apheresis column contains a matrix for affinity chromatographic removal of CRP, the matrix comprising a matrix substrate material to which ligands are bound which have the property of specifically binding CRP, the ligands bound to the matrix substrate material being selected from phosphocholine or phosphoethanolamine, or

wherein the matrix comprises a matrix substrate material functionalized with at least one ω-phosphonooxyalkylammonium group and/or at least one ω-ammoniumalkoxy-hydroxyphosphoryloxy group; and

wherein CRP binds to the matrix in the apheresis column with higher affinity than other substances present in the blood,

wherein the concentration of the alkali hydroxide in the alkali hydroxide solution is in a range of 0.04 mol/l to 0.4 mol/l.

18 . The method according to claim 17 for regeneration of an apheresis column for affinity chromatographic removal, the method characterized by the following steps:

(A) starting the redirection of the separated plasma from the plasma line into the bypass line, thereby stopping the introduction of the separated plasma from the plasma line into the apheresis column,

(B) starting the introduction of a rinsing solution via the at least one regeneration line into the plasma line or directly into the apheresis column,

(C) starting the redirection of the liquid flow exiting the apheresis column from the plasma line into the waste line,

(D) stopping the introduction of the rinsing solution and transition to the introduction of a regeneration solution via the at least one regeneration line into the plasma line or directly into the apheresis column, wherein the regeneration solution is an alkali hydroxide solution,

(E) stopping the introduction of the regeneration solution and transition to the introduction of the neutralization solution via the at least one regeneration line into the plasma line or directly into the apheresis column,

(F) stopping the introduction of the neutralization solution and transition to the introduction of the rinsing solution via the at least one regeneration line into the plasma line or directly into the apheresis column,

(G) stopping the introduction of the rinsing solution and stopping the redirection of the separated plasma from the plasma line into the bypass line, thereby direction of the separated plasma from the plasma line into the apheresis column;

(H) closing the waste line.