Characterization of a human hematopoietic progenitor cell antigen

• US 5,843,633 A
• Filed: 04/26/1996
• Issued: 12/01/1998
• Est. Priority Date: 04/26/1996
• Status: Expired due to Term

• Current Assignee: Amcell Corporation
• Original Assignee: Amcell Corporation
• Inventors: Miraglia, Sheri, Buck, David W., Yin, Amy
• Primary Examiner(s): Eisenschenk, Frank C.
• Assistant Examiner(s): RABIN, EVELYN MARIE
• Application Number:
  US08/639,891
• Time in Patent Office: 949 Days
• Field of Search: 530/391.3, 530/387.1, 530/388.1, 530/388.7, 530/389.6, 435/2, 435/7.21, 435/240.2, 435/343, 435/326, 435/346, 435/372.1
• US Class Current: 435/2
• CPC Class Codes: C07K 14/705 Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: C07K 16/28 against receptors, cell surface antigens or cell surface determinants

Abstract

An hematopoietic progenitor cell antigen, and antibodies that specifically bind to the antigen are provided. Expression of the antigen is highly tissue specific. It is only detected on a subset of hematopoietic progenitor cells derived from human bone marrow, fetal bone marrow and liver, cord blood and adult peripheral blood. The subset of cells recognized by AC133 is CD34^bright, and contains substantially all of the CFU-GM activity present in the CD34⁺ population. This highly specific distribution of AC133 makes it exceptionally useful as a reagent for isolating and characterizing human hematopoietic progenitor and stem cells. Cells selected for expression of AC133 antigen may be further purified by selection for other hematopoietic stem cell and progenitor cell markers.

Claims

1. An antibody that specifically binds to the AC133 antigen, wherein said AC133 antigen specifically binds to the antibody produced by the hybridoma cell line ATCC HB 12346.

2. An antibody according to claim 1, wherein said antibody is a monoclonal antibody, produced by a hybridoma cell line.

3. An antibody according to claim 2, wherein said monoclonal antibody blocks simultaneous binding to said AC133 antigen by the antibody produced by the hybridoma cell line ATCCHB12346.

4. An antibody according to claim 1, wherein said antibody was raised through contralateral immunization.

5. A monoclonal antibody produced by the hybridoma cell line ATCCHB12346.

8. A method according to claim 5 or 7, wherein said reagent or reagents are antibodies.

9. A method according to claim 8, wherein at least one of said antibodies is fluorochrome conjugated.

10. A method according to claim 9, wherein said selecting with said fluorochrome conjugated antibodies is by flow cytometry.

11. A method according to claim 8, wherein at least one of said antibodies is conjugated to magnetic particles.

12. A method according to claim 11, wherein said selecting with said magnetic particle conjugated antibodies is by high gradient magnetic selection.

6. A method for enrichment of hematopoietic progenitor cells, said method comprising:

combining a mixed population of human cells comprising hematopoietic progenitor cells with a reagent that specifically binds to the AC133 antigen; and

selecting for those cells that are AC133 positive;

wherein said selected cells are enriched in hematopoietic progenitor cell activity.

7. A method according to claim 6, further comprising:

combining said mixed population of human cells or said selected cell with reagents that specifically recognize at least one of the cell surface markers CDw90, CD117 and HLA-DR; and

selecting for those cells that are positive for said at least one cell surface marker.