Methods for preparing enriched human hematopoietic cell preparations

• US 6,306,575 B1
• Filed: 12/01/1995
• Issued: 10/23/2001
• Est. Priority Date: 06/16/1995
• Status: Expired due to Term

• Current Assignee: Stemcell Technologies Inc. (Stemcell Technologies Canada Inc.)
• Original Assignee: Stemcell Technologies Inc. (Stemcell Technologies Canada Inc.)
• Inventors: Thomas, Terry, Lansdorp, Peter
• Primary Examiner(s): Reeves, Julie
• Application Number:
  US08/566,295
• Time in Patent Office: 2,153 Days
• Field of Search: 530/388.7, 530/388.73, 530/388.75, 530/391.1, 530/413, 424/130.1, 424/153.1, 424/155.1, 424/173.1, 424/174.1, 424/178.1, 435/7.24, 435/2, 435/372, 435/366, 435/7.23, 435/334, 435/344, 436/503
• US Class Current: 435/2
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 16/2803 against the immunoglobulin superfamily
• CPC Class Codes: C07K 16/2806 against CD2
• CPC Class Codes: C07K 16/2809 against the T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 16/283 against Fc-receptors, e.g. CD16, CD32, CD64 CD23 C07K16/2851
• CPC Class Codes: C07K 16/289 against CD45
• CPC Class Codes: C07K 16/2896 against molecules with a "CD"-designation, not provided for elsewhere
• CPC Class Codes: C07K 16/4283 against an allotypic or isotypic determinant on Ig
• 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: C12N 5/0693 Tumour cells; Cancer cells

Abstract

The present invention relates to an antibody composition which contains antibodies specific for glycophorin A, CD3, CD24, CD16, CD14, and optionally CD45RA, CD38, CD36, CD38, CD56, CD2, CD19, CD66e, CD66b, and/or antibodies specific for antigens expressed on non-hematopoietic tumor cells. A process is also provided for enriching and recovering human hematopoietic progenitor cells and stem cells in a sample containing human hematopoietic differentiated, progenitor, and stem cells, and optionally tumor cells. The process involves reacting the sample with an antibody composition containing antibodies capable of binding to the antigens glycophorin A, CD3 CD24, CD16, and CD14, and optionally CD45RA, CD36, CD38, CD56, CD2, CD19, CD66e, CD66b, and/or antibodies specific for antigen expressed on non-hematopoietic tumor cells under conditions so that cell conjugates are formed between the antibodies and differentiated cells having the antigens glycophorin A, CD3 CD24, CD16, and CD14, and optionally CD45RA, CD38, CD36, CD38, CD56, CD2, CD19, CD66e, CD66b, and/or antibodies specific for antigens expressed on non-hematopoietic tumor cells on their surfaces. The cell conjugates are removed and a cell preparation is obtained which is enriched in human hematopoietic progenitor cells and stem cells. The invention also relates to kits for carrying out this process and to the cell preparations prepared by the process.

Claims

1. A negative selection process for enriching and recovering human hematopoietic progenitor cells and human hematopoietic stem cells in a sample containing human hematopoietic differentiated cells, human hematopoietic progenitor cells, human hematopoietic stem cells, and tumor cells comprising:

(a) reacting the sample with an antibody composition containing antibodies capable of binding to the antigens glycophorin A, CD3, CD24, CD16, and CD14, and containing an antibody specific for a non-hematopoietic antigen expressed on the tumor cells, under conditions so that antibody-cell conjugates are formed between the antibodies and cells in the sample containing the antigens glycophorin A, CD3, CD24, CD16, and CD14 and non-hematopoietic antigens on their surface;

(b) removing the conjugates; and

,

(c) recovering a cell preparation which is enriched in human hematopoietic progenitor cells and human hematopoietic stem cells, wherein the cell preparation recovered in step (c) has about a 3 to 5 log depletion of tumor cells and differentiated cells and is not labeled or coated with antibodies.

2. A process as claimed in claim 1, wherein the antibodies in the antibody composition are monoclonal antibodies.

3. A process as claimed in claim 1, wherein the antibodies in the antibody composition are labelled with a marker or are conjugated to a matrix.

4. A process as claimed in claim 1, wherein the antibodies in the antibody composition are labelled with biotin or a fluorochrome.

5. A process as claimed in claim 3, wherein the matrix is magnetic beads, a panning surface, dense particles for density centrifugation, an adsorption column, or an adsorption membrane.

6. A process as claimed in claim 5, wherein each of the antibodies in the antibody composition is a monoclonal antibody that is incorporated in a tetrameric antibody complex which comprises a first monoclonal antibody of a first animal species from the antibody composition, and a second monoclonal antibody of the first animal species which is capable of binding to at least one antigen on the surface of a matrix, which have been conjugated to form a cyclic tetramer with two monoclonal antibodies of a second animal species directed against the Fe-fragments of the antibodies of the first animal species.

7. A process as claimed in claim 1 wherein the tumor cells are from breast carcinoma, lung carcinoma, or neuroblastoma, and the antibody composition contains antibodies specific for antigens expressed on the surface of cells from breast carcinoma, lung carcinoma, or neuroblastoma.

8. A negative selection process for enriching and recovering human hematopoietic progenitor cells and human hematopoietic stem cells in a sample containing human hematopoietic differentiated cells, human hematopoietic progenitor cells, human hematopoietic stem cells, and tumor cells comprising;

(a) reacting the sample with an antibody composition containing antibodies capable of binding to the antigens glycophorin A, CD3, CD24, CD16, CD14, CD45RA, CD36, CD38, CD56, CD2, CD19, CD66e, and CD66b, and containing an antibody specific for a non-hematopoietic antigen expressed on the tumor cells, under conditions so that antibody-cell conjugates are formed between the antibodies and cells in the sample containing the antigens glycophorin A, CD3, CD24, CD16, CD14, CD45RA, CD36, CD38, CD56, CD2, CD19, CD66e, CD66b, and non-hematopoietic antigens on their surfaces;

(b) removing the conjugates; and

,

(c) recovering a cell preparation which is enriched in human hematopoietic progenitor cells and human hematopoietic stem cells, wherein the cell preparation recovered in step (c) has about a 3 to 5 log depletion of tumor cells and differentiated cells and is not labeled or coated with antibodies.

9. A process as claimed in claims 8, wherein the antibodies in the antibody composition are monoclonal antibodies.

10. A process as claimed in claim 8, wherein the antibodies in the antibody composition are labelled with a marker or the antibodies are conjugated to a matrix.

11. A process as claimed in claim 10, wherein the antibodies in the antibody composition are labelled with biotin or a fluorochrome.

12. A process as claimed in claim 10, wherein the matrix is magnetic beads, a panning surface, dense particles for density centrifugation, an adsorption column, or an adsorption membrane.

13. A process as claimed in claim 12, wherein each of the antibodies in the antibody composition is a monoclonal antibody that is incorporated in a tetrameric antibody complex which comprises a first monoclonal antibody of a first animal species from the antibody composition, and a second monoclonal antibody of the first animal species which is capable of binding to an antigen on the surface of a matrix, which have been conjugated to form a cyclic tetramer with two monoclonal antibodies of a second animal species directed against the Fc-fragments of the antibodies of the first animal species.