Materials and procedures for the purification of cells

• US 7,015,034 B2
• Filed: 10/17/2001
• Issued: 03/21/2006
• Est. Priority Date: 11/08/1996
• Status: Expired due to Fees

• Current Assignee: Morphogenesis, Inc. (RadioIO, Inc.)
• Original Assignee: Morphogenesis, Inc. (RadioIO, Inc.)
• Inventors: Lawman, Michael J. P., Lawman, Patricia
• Primary Examiner(s): SAUNDERS, DAVID A
• Application Number:
  US09/981,639
• Publication Number:
  US 20020115113A1
• Time in Patent Office: 1,616 Days
• Field of Search: 435/2, 435/7.1, 435/7.21, 435/7.24, 435/7.5, 435/30, 435/287.2, 435/372, 435/325, 436/528, 436/535, 436/806, 204/403.1
• US Class Current: 435/325
• 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 conductive immunopolymer matrix comprising:

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.

2. The conductive immunopolymer matrix 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, CD11, CD14, CD19, CD20, or CD33 determinant.

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

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

5. The conductive immunopolymer matrix a according to claim 1, wherein said polyheteroaromatic polymer is a naphthalene sulfonate-doped polypyrole or a p-toluene sulfanate-doped polypyrrole.

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

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

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

9. A conductive immunopolymer matrix comprising:

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.

10. The conductive immunopolymer matrix according to claim 9, 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.

11. The conductive immunopolymer matrix according to claim 9, wherein said antibody bound to said Fc receptor of said second immunopolymer layer binds to a CD34 determinant.

12. The conductive immunopolymer matrix according to claim 9, wherein said polyphenol polymer has a net negative charge at the surface of said polymer.

13. The conductive immunopolymer matrix according to claim 9, wherein said conductive immunopolymer matrix is provided in a capillary, spiral sheet, or parallel sheet.

14. The conductive immunopolymer matrix according to claim 9, wherein said first and second immunopolymer layers are sequentially oriented with respect to each other.

15. A conductive immunopolymer matrix comprising a polymer and a molecule having binding specificity for a target molecule, wherein said polymer comprises a polyphenol polymer, and wherein said molecule having binding specificity for a target molecule is an Fc receptor and is entrapped within said polymer.

16. The conductive immunopolymer matrix according to claim 15, wherein an antibody is bound to said Fc receptor.

17. The conductive immunopolymer matrix according to claim 16, wherein said antibody binds to a CD34 determinant.

18. The conductive immunopolymer matrix according to claim 16, wherein said antibody binds to a CD3, CD4, CD7, CD8, CD10, CD11b, CD14, CD19, CD20, or CD33 determinant.

19. The conductive immunopolymer matrix according to claim 15, wherein said polyphenol polymer has a net negative charge at the surface of said polymer.

20. The conductive immunopolymer matrix according to claim 15, wherein said conductive immunopolymer matrix is provided in a capillary, spiral sheet, or parallel sheet.

21. The conductive immunopolymer matrix according to claim 15, 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.

22. The conductive immunopolymer matrix according to claim 15, wherein said conductive immunopolymer matrix comprises multiple layers of said polymer, and wherein a different antibody is bound to said Fc receptor in each of said layers.