Claims
- 1. Agonist molecules which specifically bind to or interact with human G-CSF receptor to stimulate cell proliferation and differentiation.
- 2. The agonist molecules of claim 1 which stimulate proliferation and differentiation of neutrophils or its progenitor cells.
- 3. Agonist molecules which specifically bind to or interact with human G-CSF receptor and dimerize the receptor or activate phosphorylation of kinases associated with the receptor to stimulate cell proliferation and differentiation.
- 4. The human G-CSF receptor of claim 1 which is a native human G-CSF receptor, or its mutants with substitutions, insertions or deletions.
- 5. The agonist molecules of claims 1 or 3 which interact with the extracellular portion of human G-CSF receptor.
- 6. The agonist molecules of claim 5 which interact at a region between amino acid residues 1-603 (SEQ ID NO: 27) of the G-CSF receptor.
- 7. The extracellular portion of human G-CSF receptor of claim 5 which is the extracellular portion of human G-CSF receptor of claim 4.
- 8. The agonist molecules of claims 1 or 3 which are monoclonal antibodies, or fragments, homologues or analogues thereof, or peptides or organic compounds.
- 9. The fragments of claim 6 which are F(ab)′2, Fab or scFv.
- 10. The monoclonal agonist antibodies of claim 8 which include mAb163-93 and mAb174-74-11.
- 11. Agonist molecules which bind to the same epitope as either the monoclonal antibody mAb163-93 or mAb 174-24-11.
- 12. Agonist molecules of claims 1 or 3 which are capable of stimulating the proliferation of human or mouse cells expressing the human G-CSF receptor as determined in an in vitro proliferation assay.
- 13. The monoclonal agonist antibody mAb163-93 of claim 10, which belongs to IgG1 subclass, wherein the CDRs of the variable region heavy chain include one or more of the following amino acid sequences:
CDR1: Asn Tyr Gly Met Asn (SEQ ID NO: 15) CDR2: Trp Ile Asn Thr Tyr Thr Gly Glu Pro Thr Tyr Ala Gly Asp Phe Lys Gly (SEQ ID NO: 16) CDR3: Glu Gly Phe Tyr Gly Gly His Pro Gly Phe Asp Tyr (SEQ ID NO: 17) Or the CDRs of the variable region light chain include one or more of the following amino acid sequences: CDR1: Lys Ser Ser Gln Ser Leu Leu Ser Ser Arg Thr Arg Lys Asn Tyr Leu Ala (SEQ ID NO: 18) CDR2: Trp Ala Ser Thr Arg Glu Ser (SEQ ID NO: 19) CDR3: Lys Gln Ser Tyr Asn Leu Arg Thr (SEQ ID NO: 20)
- 14. The monoclonal agonist antibody mAb174-74-11 of claim 10, which belongs to IgG2a subclass, wherein the CDRs of the variable region heavy chain include one or more of the following amino acid sequences:
CDR1: Ser Tyr Ala Met Ser (SEQ ID NO: 21) CDR2: Gly Ile Ser Ser Gly Gly Ser Tyr Ser Tyr Tyr Pro Gly Thr Leu Lys Gly (SEQ ID NO: 22) CDR3: Glu Ala Tyr Asn Asn Tyr Asp Ala Leu Asp Tyr (SEQ ID NO: 23) or CDRs of the light chain variable region include one or more of the following amino sequences: CDR1: Arg Ala Ser Ser Ser Val Thr Tyr Val His (SEQ ID NO: 24) CDR2: Ala Thr Ser Asn Leu Ala Ser (SEQ ID NO: 25) CDR3: Gln Gln Trp Thr Ser Asn Pro Phe Thr (SEQ ID NO: 26)
- 15. Hybridoma cell lines producing the monoclonal agonist antibodies, fragments, homologues, analogues or peptides of claim 8.
- 16. The hybridoma cell lines of claim 13 which are hybridoma cell lines 163-93 or 174-74-11.
- 17. DNA sequences encoding monoclonal agonist antibodies, fragments, homologues, analogues or peptides of claim 8.
- 18. Gene delivery systems including DNA sequences encoding monoclonal agonist antibodies, fragments, homologues, analogues or peptides of claim 8.
- 19. Gene delivery systems including one or more of the DNA sequences shown in SEQ ID NOS: 15 through 26.
- 20. Gene delivery systems of claim 16 wherein said systems include viral vectors, plasmids, or non-vector delivery systems.
- 21. A method of treating neutropenia comprising administering the molecules of any of claims 1-3.
- 22. A method of treating neutropenia comprising administering the molecules of claim 5.
- 23. A method of treating neutropenia comprising administering the molecules of claim 8.
- 24. A method of screening a molecule for G-CSF agonist activity comprising determining if the molecule can stimulate the proliferation of G-CSF-dependent cells.
- 25. The method of claim 22 wherein the measurement is made using an in vitro assay.
- 26. The method of claim 23 wherein the assay method is an MTT-based calorimetric assay or an 3H-thymidine uptake assay using cells expressing G-CSF receptor.
- 27. The G-CSF dependent cells of claim 22 which express proteins, or fragments thereof, having the sequence of the extracellular domain or its part of the G-CSF receptor on its cell membrane surface.
- 28. A method for dectecting the human G-CSF receptor immunologically by means of antibodies of claim 8.
- 29. A method for immunological dectection of a cell expressing the human G-CSF receptor on the cell surface by means of the antibodies of claim 8.
- 30. A method for detecting and determining a soluble human F-CSF receptor immunologically by means of antibodies of claim 8.
RELATED APPLICATION
[0001] This application claims priority to U.S. Provisional Application Serial No. 60/083,575, filed on Apr. 30, 1998.
Provisional Applications (1)
|
Number |
Date |
Country |
|
60083575 |
Apr 1998 |
US |
Continuations (1)
|
Number |
Date |
Country |
Parent |
09303155 |
Apr 1999 |
US |
Child |
10071962 |
Feb 2002 |
US |