METHODS FOR TREATING A SUSPENSION OF MONONUCLEAR CELLS TO FACIILTATE EXTRACORPOREAL PHOTOPHERESIS

• US 20140370491A1
• Filed: 06/13/2013
• Published: 12/18/2014
• Est. Priority Date: 06/13/2013
• Status: Active Grant

• Current Assignee: Fenwal Incorporated (Fresenius SE & Co. KGaA)
• Original Assignee: Fenwal Incorporated (Fresenius SE & Co. KGaA)
• Inventors: Radwanski, Katherine
• Granted Patent: US 10,213,544 B2
• Time in Patent Office: Days
• US Class Current: 435/2
• CPC Class Codes: A61M 1/3496 Plasmapheresis; Leucopheresis; Lymphopheresis A61M1/3472 takes precedence
• CPC Class Codes: A61M 1/36225 with blood pumping means or components thereof
• CPC Class Codes: A61M 1/3681 by irradiation
• CPC Class Codes: A61M 1/3683 using photoactive agents
• CPC Class Codes: A61M 1/3686 by removing photoactive agents after irradiation
• CPC Class Codes: A61M 1/3692 Washing or rinsing blood or blood constituents
• CPC Class Codes: A61M 1/3693 using separation based on different densities of components, e.g. centrifuging
• CPC Class Codes: A61M 1/3696 with means for adding or withdrawing liquid substances during the centrifugation, e.g. continuous centrifugation
• CPC Class Codes: A61M 2202/0439 White blood cells; Leucocytes lymphocytes A61M2202/0407
• CPC Class Codes: A61M 2205/053 ultraviolet

Abstract

A method for performing a photopheresis procedure is provided comprising collecting MNCs in a suspension comprising RBCs and plasma and lysing the red blood cells in the solution, preferably by combining the suspension with a solution to cause lysis. In one example, the solution for causing lysis of the red blood cells comprises ammonium chloride, and the suspension including the ammonium chloride is incubated to cause lysing. After lysing, the suspension may be washed to remove plasma and hemoglobin freed by the lysis of the red blood cells, and an ultraviolet light activated substance is added to the suspension. The suspension is then irradiated with ultraviolet light.

Claims

1. A method of extracorporeal photopheresis of mononuclear cells comprising:

collecting a suspension of mononuclear cells containing red blood cells;

adding a lysing agent to the suspension;

incubating the suspension to lyse the red blood cells;

removing from the suspension hemoglobin freed by the lysing of the red blood cells;

adding an ultraviolet light activated substance to the suspension; and

irradiating the suspension with ultraviolet light.

3. The method of claim 1 wherein the lysing agent does not substantially adversely affect the viability of the mononuclear cells in the suspension.

4. The method of claim 1 in which the lysing agent comprises ammonium chloride.

5. The method of claim 4 wherein the ammonium chloride is added in an aqueous solution at predetermined concentration.

6. The method of claim 5 wherein the predetermined concentration is from about 100 mM to 200 mM.

7. The method of claim 1 in which the suspension includes plasma and the method includes removing plasma from the suspension.

8. The method of claim 7 wherein the plasma and hemoglobin are removed substantially simultaneously.

9. The method of claim 8 wherein the removing includes washing.

11. The method of claim 1 in which removing hemoglobin is accomplished by centrifuging the solution.

12. The method of claim 1 in which the suspension is irradiated with ultraviolet light for five minutes or less.

13. The method of claim 1 in which the ultraviolet light activated substance comprises psoralen.

14. The method of claim 13 wherein the ultraviolet light activated substance comprises 8-methoxypsoralen.

15. The method of claim 1 in which the suspension is irradiated with UVA ultraviolet light.

2. (canceled)

10. (canceled)

16. A method for performing extracorporeal photopheresis of mononuclear cells comprising:

collecting mononuclear cells in a suspension comprising red blood cells and plasma;

combining the suspension with a lysing solution;

incubating the suspension to lyse the red blood cells;

removing from the suspension plasma and hemoglobin freed by the lysis of the red blood cells;

adding an ultraviolet light activated agent to the suspension; and

irradiating the suspension with ultraviolet light.

17. The method of claim 16 wherein removing plasma and hemoglobin freed by the lysis of the red blood cells is accomplished by washing the suspension by centrifugation.

18. The method of claim 16 wherein the lysing solution comprises ammonium chloride.

19. A method of extracorporeal photopheresis of mononuclear cells in a suspension of concentrated mononuclear cells that includes red blood cells and plasma, comprising:

removing sufficient red blood cells from the suspension to substantially decrease absorption of ultraviolet light by non-target substances in the suspension;

adding an ultraviolet light activated substance to the suspension; and

irradiating the suspension with ultraviolet light.

20. The method of claim 19 wherein the red blood cells are removed by either immunomagnetic cell separation or density gradient separation of the suspension.