Materials and procedures for the purification of cells

• US 20060105399A1
• Filed: 12/20/2005
• Published: 05/18/2006
• Est. Priority Date: 11/08/1996
• Status: Active Grant

• Current Assignee: Michael Lawman, Patricia Lawman
• Original Assignee: Michael Lawman, Patricia Lawman
• Inventors: Lawman, Michael, Lawman, Patricia
• Granted Patent: US 7,598,081 B2
• Time in Patent Office: Days
• US Class Current: 435/7.200
• CPC Class Codes: C07K 16/2896 against molecules with a "CD"-designation, not provided for elsewhere
• CPC Class Codes: C07K 17/08 the carrier being a synthetic polymer
• CPC Class Codes: C12N 5/0087 Purging against subsets of blood cells, e.g. purging alloreactive T cells
• CPC Class Codes: C12N 5/0647 Haematopoietic stem cells; Uncommitted or multipotent progenitors
• CPC Class Codes: Y10S 436/806 Electrical property or magnetic property

Abstract

The subject invention provides new materials and methods for the efficient isolation and purification of stem cells. Specifically, conductive immunopolymers with stem cell specific antibodies can be used to remove stem cells from biological fluids.

Claims

1. A method for isolating a target cell from a sample, said method comprising i) contacting said sample with a conductive immunopolymer matrix, wherein said conductive immunopolymer matrix comprises:

a) a first immunopolymer layer comprising a polyheteroaromatic polymer and an Fc receptor entrapped within said polymer, wherein a first antibody is bound to said Fc receptor in said first polymer; and

b) a second immunopolymer layer comprising a polyheteroaromatic polymer and an Fc receptor entrapped within said polymer, wherein a second antibody is bound to said Fc receptor in said second polymer and said second antibody binds to a determinant or antigen that said first antibody does not bind; and

wherein said first and second immunopolymer layers further comprise a cell monitoring system, wherein said cell monitoring system comprises horseradish peroxidase and glucose oxidase entrapped within said first and second immunopolymers; and

wherein said first or second antibody binds to a molecule expressed on said target cell, thereby isolating said target cell from said sample.

2. The method according to claim 1, wherein said antibody bound to said Fc receptor of said first immunopolymer layer binds to a CD3, CD4, CD7, CD8, CD10, CD11b, CD14, CD19, CD20, or CD33 determinant.

3. The method according to claim 1, wherein said antibody bound to said Fc receptor of said second immunopolymer layer binds to a CD34 determinant.

4. The method according to claim 1, wherein said polyheteroaromatic polymer is an alkyl substituted polythiophene or a polypyrrole.

5. The method according to claim 1, wherein said polyheteroaromatic polymer is a naphthalene sulfonate-doped polypyrrole or a p-toluene sulfonate-doped polypyrrole.

6. The method according to claim 1, wherein said polyheteroaromatic polymer has a net negative charge at the surface of said polymer.

7. The method according to claim 1, wherein said conductive immunopolymer matrix is provided in a capillary, spiral sheet, or parallel sheet.

8. The method according to claim 1, wherein said first and second immunopolymer layers are sequentially oriented with respect to each other.

9. The method according to claim 1, wherein said target cell is a tumor cell, a stem cell, a blood cell, or a bone marrow cell.

10. The method according to claim 1, wherein said method further comprises washing said conductive immunopolymer matrix after step (i).

11. The method according to claim 1, wherein said method further comprises releasing said first or second antibody from said first or second immunopolymer layer after step (i).

12. A method for isolating a target cell from a sample, said method comprising

i) contacting said sample with a conductive immunopolymer matrix, wherein said conductive immunopolymer matrix comprises:

a) a first immunopolymer layer comprising a polyphenol polymer and an Fc receptor entrapped within said polymer, wherein a first antibody is bound to said Fc receptor in said first polymer; and

b) a second immunopolymer layer comprising a polyphenol polymer and an Fc receptor entrapped within said polymer, wherein a second antibody is bound to said Fc receptor in said second polymer and said second antibody binds to a determinant or antigen that said first antibody does not bind; and

wherein said first and second immunopolymer layers further comprise a cell monitoring system, wherein said cell monitoring system comprises horseradish peroxidase and glucose oxidase entrapped within said first and second immunopolymers; and

wherein said first or second antibody binds to a molecule expressed on said target cell, thereby isolating said target cell from said sample.

13. The method according to claim 12, wherein said antibody bound to said Fc receptor of said first immunopolymer layer binds to a CD3, CD4, CD7, CD8, CD10, CD11b, CD14, CD19, CD20, or CD33 determinant.

14. The method according to claim 12, wherein said antibody bound to said Fc receptor of said second immunopolymer layer binds to a CD34 determinant.

15. The method according to claim 12, wherein said polyphenol polymer has a net negative charge at the surface of said polymer.

16. The method according to claim 12, wherein said conductive immunopolymer matrix is provided in a capillary, spiral sheet, or parallel sheet.

17. The method according to claim 12, wherein said first and second immunopolymer layers are sequentially oriented with respect to each other.

18. The method according to claim 12, wherein said target cell is a tumor cell, a stem cell, a blood cell, or a bone marrow cell.

19. The method according to claim 12, wherein said method further comprises washing said conductive immunopolymer matrix after step (i).

20. The method according to claim 12, wherein said method further comprises releasing said first or second antibody from said first or second immunopolymer layer after step (i).

21. A method for isolating a target cell from a sample, said method comprising

(i) contacting said sample with a conductive immunopolymer matrix, wherein said conductive immunopolymer matrix comprises a polymer and an antibody, wherein said polymer comprises a polyphenol polymer, and wherein said antibody binds to a molecule expressed on said target cell, thereby isolating said target cell from said sample.

22. The method according to claim 21, wherein said antibody is bound to an Fc receptor in said polymer.

23. The method according to claim 21, wherein said polyphenol polymer has a net negative charge at the surface of said polymer.

24. The method according to claim 21, wherein said antibody binds to a CD34 determinant.

25. The method according to claim 21, wherein said antibody binds to a CD3, CD4, CD7, CD8, CD10, CD11b, CD14, CD 19, CD20, or CD33 determinant.

26. The method according to claim 21, wherein said conductive immunopolymer matrix is provided in a capillary, spiral sheet, or parallel sheet.

27. The method according to claim 21, wherein said conductive immunopolymer matrix further comprises a cell monitoring system, wherein said cell monitoring system comprises horseradish peroxidase and glucose oxidase entrapped within said polymer.

28. The method according to claim 21, wherein said conductive immunopolymer matrix comprises multiple layers of said polymer, and wherein each layer of said polymer comprises a different antibody.

29. The method according to claim 21, wherein said target cell is a tumor cell, a stem cell, a blood cell, or a bone marrow cell.

30. The method according to claim 21, wherein said method further comprises washing said conductive immunopolymer matrix after step (i).

31. The method according to claim 21, wherein said method further comprises releasing said antibody from said polymer after step (i).