Methods and compositions for separating rare cells from fluid samples
The present invention includes methods of enriching rare cells, such as cancer cells, from biological samples, such as blood samples. The methods include performing at least one debulking step on a blood sample and selectively removing at least one type undesirable component from the blood sample to obtain a blood sample that is enriched in a rare cell of interest. In some embodiments magnetic beads coupled to specific binding members are used to selectively removed components.
1. A method of enriching rare cells from a blood sample, comprising:
a) performing at least one debulking step on a blood sample; and
b) selectively removing at least one type of undesirable component from said blood sample to enrich a rare cell of interest in said blood sample, wherein said at least one type of undesirable component is a white blood cell;
wherein selectively removing the white blood cell from said blood sample comprises contacting said blood sample with an antibody or an antibody fragment that selectively binds to the white blood cell to be removed.
2. The method according to claim 1 , wherein said antibody or antibody fragment is coupled to a solid support.
3. The method of claim 2 , wherein said antibody or antibody fragment selectively binds to CD3, CD11b, CD14, CD17, CD31, CD45, CD50, CD53, CD63, CD69, CD81, CD84, CD102 or CD166.
4. The method of claim 3 , wherein said antibody or antibody fragment selectively binds to CD50.
5. The method according to claim 1 , wherein said blood sample is a serum-depleted blood sample or a preserved blood sample.
6. The method according to claim 1 , wherein said blood sample is a peripheral blood sample.
7. The method according to claim 1 , wherein said debulking step is a centrifugation step.
8. The method according to claim 1 , wherein the debulking step removes red blood cells (RBC).
9. The method according claim 8 , wherein red blood cells are removed by a method selected from the group consisting of sedimentation, lysis, or binding to a red blood cell specific binding member.
10. The method according to claim 1 , wherein said antibody or antibody fragment is coupled to a magnetic bead, further wherein the white blood cell is removed from the sample by using a magnet to capture said magnetic bead to which said antibody or antibody fragment is coupled and bound.
11. The method according to claim 10 , wherein said antibody or antibody fragment selectively binds to CD50.
12. The method according to claim 1 , further comprising labeling said rare cells.
13. The method according to claim 12 , wherein said rare cells are cancer cells or other non-hematopoietic cells.
14. The method according to claim 13 , further comprising contacting said blood sample with a specific binding member that selectively binds to a second type of said undesirable component, wherein said blood sample is enriched by removal of said second type of undesirable component.
15. The method of claim 14 , wherein said specific binding member is an antibody that selectively binds to CD31, CD36, CD41, CD42 (a, b or c), CD51, or CD51/61.
16. The method according to claim 11 , further comprising labeling said rare cells.
17. The method according to claim 16 , wherein said rare cells are cancer cells or other non-hematopoietic cells.
18. The method according to claim 17 , further comprising contacting said blood sample with a specific binding member that selectively binds to a second type of said undesirable component, wherein said blood sample is enriched by removal of said second type of undesirable component.
19. The method of claim 18 , wherein said specific binding member is an antibody that selectively binds to CD31, CD36, CD41, CD42 (a, b or c), CD51, or CD51/61.
20. A method for enriching cancer cells from a blood sample, comprising:
a) removing red blood cells from the blood sample by lysis of said red blood cells or by selective sedimentation of said red blood cells;
b) removing white blood cells from said blood sample by contacting said blood sample with magnetic beads to which a selective binding member is coupled, wherein said selective binding member is an antibody that specifically binds to CD50; and
c) labeling at least one type of cancer cells.
21. The method of claim 20 , further comprising a step of analyzing said at least one type of cancer cells using a method selected from the group consisting of spectral imaging, fluorescence microscopy, visible light microscopy, and manual or automated image analysis.
22. The method of claim 20 , further comprising isolating said at least one type of cancer cells by fluorescence activated cell sorting or laser cytometry.