Materials and procedures for the purification of cells

• US 8,030,069 B2
• Filed: 04/24/2009
• Issued: 10/04/2011
• 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 D.
• Primary Examiner(s): Saunders; David A
• Application Number:
  US12/429,750
• Publication Number:
  US 20090221074A1
• Time in Patent Office: 893 Days
• Field of Search: None
• 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 polymer matrix comprising a molecule having binding specificity for a target molecule, wherein said molecule having binding specificity for a target molecule is bound to an Fc receptor, Protein G, or avidin molecule incorporated in said conductive polymer matrix, wherein said molecule having binding specificity for a target molecule can be released from said conductive polymer matrix upon application of an electric current.

2. The conductive polymer matrix according to claim 1, wherein said molecule having binding specificity for a target molecule is an antibody, or an antigen binding fragment thereof.

3. The conductive polymer matrix according to claim 2, wherein said antibody is a monoclonal antibody.

4. The conductive polymer matrix according to claim 2, wherein said antibody binds to a CD3, CD4, CD7, CD8, CD10, CD11b, CD14, CD19, CD20, CD33, or CD34 determinant.

5. The conductive polymer matrix according to claim 1, wherein said molecule having binding specificity for a target molecule comprises a biotin moiety.

6. The conductive polymer matrix according to claim 1, wherein said conductive polymer matrix comprises polyheteroaromatic polymer, polyphenol polymer, polyacetylene polymer, polyparaphenylene polymer, or polyphenylenevinylene polymer.

7. The conductive polymer matrix according to claim 6, wherein said polyheteroaromatic polymer is an alkyl-substituted polythiophene or polypyrrole.

8. The conductive polymer matrix according to claim 1, 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.

9. A method for isolating a target cell from a sample comprising a mixture of cells, said method comprising:

contacting said mixture of cells with a conductive polymer matrix, wherein said conductive polymer matrix comprises a molecule having binding specificity for a target molecule expressed on said target cell, wherein said molecule having binding specificity for a target molecule is bound to an Fc receptor, Protein G, or avidin molecule incorporated in said conductive polymer matrix, wherein said molecule having binding specificity for a target molecule can be released from said conductive polymer matrix upon application of an electric current;

binding said target cell to said molecule having binding specificity for said target cell; and

releasing said molecule having binding specificity for said target molecule from said conductive polymer matrix.

10. The method according to claim 9, wherein said method further comprises washing said polymer matrix to remove unbound material.

11. The method according to claim 9, wherein said molecule having binding specificity for said target molecule is an antibody, or an antigen binding fragment thereof.

12. The method according to claim 11, wherein said antibody is a monoclonal antibody.

13. A device for isolating a target cell from a mixture of cells, said device comprising a cathode and an anode for passing an electric current, and a conductive polymer matrix comprising a molecule having binding specificity for a target molecule, wherein said molecule having binding specificity for a target molecule is bound to an Fc receptor, Protein G, or avidin molecule incorporated in said conductive polymer matrix, wherein said molecule having binding specificity for a target molecule can be released from said conductive polymer matrix upon application of an electric current.

14. The device according to claim 13, wherein said molecule having binding specificity for a target molecule is an antibody, or an antigen binding fragment thereof.

15. The device according to claim 14, wherein said antibody is a monoclonal antibody.

16. The device according to claim 14, wherein said antibody binds to a CD3, CD4, CD7, CD8, CD10, CD11 b, CD14, CD19, CD20, CD33, or CD34 determinant.

17. The device according to claim 13, wherein said molecule having binding specificity for a target molecule comprises a biotin moiety.

18. The device according to claim 13, wherein said conductive polymer matrix comprises polyheteroaromatic polymer, polyphenol polymer, polyacetylene polymer, polyparaphenylene polymer, or polyphenylenevinylene polymer.

19. The device according to claim 18, wherein said polyheteroaromatic polymer is an alkyl-substituted polythiophene or polypyrrole.

20. The device according to claim 13, 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.