Monoclonal antibodies and their use
Isolated monoclonal antibodies are disclosed herein that specifically bind a cell surface antigen expressed on the human pancreatic endocrine cells or a subset thereof, and/or a precursor thereof. Isolated monoclonal antibodies are also disclosed herein that specifically bind a cell surface antigen expressed on human pancreatic exocrine cells or human ductal cells. Humanized forms of these antibodies, and functional fragments of these antibodies, are also disclosed. The antibodies can be conjugated to an effector molecule, such as a detectable marker, a therapeutic agent, or a toxin. These antibodies are of use to detect and isolate pancreatic cells or a subset thereof. The antibodies can be used for in vitro or in vivo detection and/or isolation of pancreatic endocrine cells. Methods of treating a pancreatic tumor are also disclosed. In several examples, the isolated monoclonal antibodies bind pancreatic endocrine cells and can be used to detect diabetes or a pancreatic endocrine cell tumor.
1. An isolated monoclonal antibody, comprising
a heavy chain comprising an H-CDR1, H-CDR2 and H-CDR3 of a monoclonal antibody produced by a HIC1 2-B4, hybridoma; and a light chain comprising an L-CDR-1, L-CDR2 and L-CDR3 of a monoclonal antibody produced by the HIC1-2-B4 hybridoma, wherein the monoclonal antibody specifically binds pancreatic endocrine cells.
2. The isolated monoclonal antibody of claim 1 , wherein the monoclonal antibody is produced by the HIC1 2 B4 hybridoma.
3. The isolated monoclonal antibody of claim 1 , comprising a human framework region.
4. An isolated functional antigen-binding fragment of the isolated monoclonal antibody of claim 1 , wherein the antigen binding fragment specifically binds pancreatic endocrine cells.
5. The isolated antigen-binding fragment of the monoclonal antibody of claim 4 , wherein the antigen-binding fragment is a scFV, a Fv, a Fab, a F(ab′) 2 or a scFV 2 fragment.
6. The isolated monoclonal antibody of claim 1 , or an antigen-binding fragment thereof, conjugated to an effector molecule.
7. The isolated monoclonal antibody or antigen-binding fragment of claim 6 , wherein the effector molecule is a detectable marker or a toxin.
8. The isolated monoclonal antibody or antigen-binding fragment of claim 7 , wherein the detectable marker is a fluorescent marker or a radiolabel.
9. The isolated antibody or antigen-binding fragment of claim 7 , wherein the toxin comprises ricin A, abrin, diphtheria toxin or a subunit thereof, Pseudomonas exotoxin or a portion thereof, saporin, restrictocin or gelonin.
10. An isolated nucleic acid encoding the monoclonal antibody of claim 1 , or encoding an antigen binding fragment of the monoclonal antibody.
11. The isolated nucleic acid of claim 10 , operably linked to a promoter.
12. An expression vector comprising the nucleic acid of claim 10 .
13. An isolated host cell transformed with the expression vector of claim 12 .
14. A hybridoma producing the monoclonal antibody of claim 1 .
15. A composition comprising an effective amount of the monoclonal antibody of claim 1 or an antigen-binding fragment thereof and physiologically acceptable carrier.
16. A method for detecting a pancreatic endocrine cell, comprising:
contacting a cell of interest with the monoclonal antibody of claim 1 or an antigen binding fragment thereof; and
detecting binding of the monoclonal antibody or the antigen binding fragment thereof to the cell,
wherein binding of the monoclonal antibody or the antigen binding fragment thereof to the cell indicates that the cell is a pancreatic endocrine cell.
17. The method of claim 16 , wherein the pancreatic endocrine cell is in vitro.
18. The method of claim 16 , wherein the pancreatic endocrine cell in vivo.
19. The method of claim 17 , wherein the pancreatic endocrine cell is isolated from a subject that has a pancreatic endocrine cell tumor or diabetes.
20. A method of isolating a pancreatic endocrine cell, comprising:
contacting a suspension of pancreatic cells with the monoclonal antibody of claim 1 or an antigen binding fragment thereof to form an immune complex; and
isolating the immune complex, thereby isolating the pancreatic endocrine cell.
21. The method of claim 20 , further comprising contacting the immune complex with a second antibody that specifically binds the monoclonal antibody of claim 3 or the antigen-binding fragment thereof.
22. The method of claim 20 , wherein the monoclonal antibody of claim 3 or the antigen-biding fragment thereof is coupled to a detectable marker.
23. The method of claim 21 , wherein the second antibody is coupled to a detectable marker.
24. The method of claim 22 , wherein the detectable marker is a fluorescent marker.
25. A method for isolating a pancreatic endocrine cell, comprising:
contacting cells with the antibody of claim 1 ;
detecting binding of the antibody to the cells; and
isolating a cell bound to the antibody,
thereby isolating the pancreatic endocrine cell, wherein the pancreatic endocrine cell produces insulin or glucagon.
26. The method of claim 25 , wherein the cells are from a human cadaveric islet.
27. The method of claim 25 , wherein the cells are differentiated from an embryonic stem cell, an adult stem cell or a progenitor cell.
28. The method of claim 16 , wherein the cell is from a human cadaveric islet.
29. The method of claim 16 , wherein the cell is differentiated from an embryonic stem cells, an adult stem cell or a progenitor cell.
30. A method for determining the purity of a human cadaveric islet preparation comprising:
contacting a cell of interest with the monoclonal antibody of claim 1 or an antigen-binding fragment thereof; and
detecting binding of the monoclonal antibody or the antigen-binding fragment thereof,
wherein the extent of binding of the monoclonal antibody or the antigen binding fragment thereof to the cadraveric islet preparation indicates the purity of the islet preparation.
31. The method of claim 30 , further comprising assessing insulin secretion of the islet preparation.
32. The method of claim 30 , wherein the method assesses the suitability of the islet cell preparation for transplantation.