Claims for Patent: 6,689,566

Summary for Patent: 6,689,566

Title:	Methods, kits, and antibodies for detecting parathyroid hormone
Abstract:	The present invention relates to novel methods and devices for detecting whole or non-fragmented parathyroid hormone (wPTH) in a biological sample. In particular, a novel monoclonal or polyclonal antibody or antibody fragment is used that is specific for a portion of the initial peptide sequence for wPTH which comprises a domain for adenylate cyclase activation, (amino acids 2 to 8), wherein at least four amino acids in this sequence are part of the reactive portion with the antibody. The present wPTH assay can differentiate between the complete 1 to 84 amino acid sequence form of PTH and a large but incomplete 7 to 84 amino acid sequence form of PTH that is measured by commercially available \"intact\" or I-PTH assays. Measurement of both the complete biologically active form of PTH and the large but biologically inactive form of PTH can lead to the misdiagnosis of the parathyroid function in patients.
Inventor(s):	Cantor; Thomas L. (El Cajon, CA), Gao; Ping (San Diego, CA)
Assignee:	Scantibodies Laboratory, Inc. (Santee, CA)
Application Number:	09/231,422
Patent Claims:	1. A substantially pure antibody or antibody fragment specific for an initial peptide sequence of whole parathyroid hormone wherein said initial peptide sequence consists of VAL-SER-GLU-ILE-GLN-LEU-MET (SEQ ID NO:3), and wherein at least four amino acids in said initial peptide sequence are part of a reactive portion with said antibody. 2. The antibody of claim 1, which is a monoclonal antibody. 3. The antibody of claim 1, which is a polyclonal antibody. 4. The antibody of claim 1, which antibody specifically binds to a parathyroid hormone epitope selected from the group consisting of amino acid residues 2-5, 3-6, 4-7, 5-8, 2-6, 3-7 and 4-8 of human parathyroid hormone (SEQ ID NO:2). 5. A method for measuring an amount of whole parathyroid hormone in a sample comprising: a) adding to a sample a labeled antibody or antibody fragment specific for an initial peptide sequence of whole parathyroid hormone wherein said initial peptide sequence consists of VAL-SER-GLU-ILE-GLN-LEU-MET (SEQ ID NO:3), and wherein at least four amino acids in said initial peptide sequence are part of a reactive portion to said labeled antibody; b) allowing said labeled antibody to bind to whole parathyroid hormone present, thereby forming a complex; and c) measuring the amount of said labeled complex to measure the amount of whole parathyroid hormone in said sample while not detecting an interfering non-(1-84) parathyroid hormone fragment. 6. The method of claim 5, wherein the labeled anti-parathyroid hormone antibody or antibody fragment is a monoclonal antibody. 7. The method of claim 5, wherein the labeled anti-parathyroid hormone antibody or antibody fragment is a polyclonal antibody. 8. The method of claim 5, wherein a second antibody is added which is bound to a solid support and specifically binds to a portion of whole parathyroid hormone other than the initial peptide sequence which binds to the labeled antibody. 9. The method of claim 8, wherein the solid support is selected from the group consisting of a protein binding surface, colloidal metal particles, iron oxide particles, latex particles and polymeric beads. 10. The method of claim 9, wherein the complex precipitates from solution. 11. The method of claim 5, wherein the label of the labeled antibody is selected from the group consisting of a chemiluminescent agent, a colorimetric agent, an energy transfer agent, an enzyme, a fluorescent agent and a radioisotope. 12. The method of claim 5, wherein the labeled antibody specifically binds to a parathyroid hormone epitope selected from the group consisting of amino acid residues 2-5, 3-6, 4-7, 5-8, 2-6, 3-7 and 4-8 of human parathyroid hormone (SEQ ID NO:2). 13. A method for measuring an amount of whole parathyroid hormone in a sample comprising: a) adding to a sample a first antibody or antibody fragment specific for an initial peptide sequence of whole parathyroid hormone wherein said initial peptide sequence consists of VAL-SER-GLU-ILE-GLN-LEU-MET (SEQ ID NO:3), and wherein at least four amino acids in said initial peptide sequence are part of a reactive portion to said first antibody; b) allowing said first antibody to bind to whole parathyroid hormone present, thereby forming a complex; c) labeling said complex by adding a second labeled antibody that specifically binds to a portion of whole parathyroid hormone other than said initial peptide sequence that binds to said first antibody to form a labeled complex; and d) measuring the amount of said labeled complex to measure the amount of whole parathyroid hormone in said sample while not detecting an interfering non-(1-84) parathyroid hormone fragment. 14. The method of claim 13, wherein the second labeled antibody is added sequentially or simultaneously with the first antibody. 15. The method of claim 13, wherein the first antibody is bound to a solid support. 16. The method of claim 13, further comprising binding a third antibody to an epitope left open after the whole parathyroid hormone binds to the first antibody and the second antibody, thereby forming a precipitating mass. 17. The method of claim 13, wherein the first antibody specifically binds to a parathyroid hormone epitope selected from the group consisting of amino acid residues 2-5, 3-6, 4-7, 5-8, 2-6, 3-7 and 4-8 of human parathyroid hormone (SEQ ID NO:2). 18. A method for measuring whole parathyroid hormone by a precipitating or turbidometric immunoassay comprising: a) adding to a sample an antibody or antibody fragment specific for an initial peptide sequence of whole parathyroid hormone wherein said initial peptide sequence consists of VAL-SER-GLU-ILE-GLN-LEU-MET (SEQ ID NO:3), and wherein at least four amino acids in said initial peptide sequence are part of an antibody reactive portion of said peptide, said antibody being attached to a colloidal particle or moiety which can be used to detect a signal change; b) allowing said antibody to bind to whole parathyroid hormone present, thereby forming a complex; and c) measuring change in said signal due to the formation of said complex to measure whole parathyroid hormone in said sample while not detecting an interfering non-(1-84). parathyroid hormone fragment. 19. The method of claim 18, wherein the first antibody specifically binds to a parathyroid hormone epitope selected from the group consisting of amino acid residues 2-5, 3-6, 4-7, 5-8, 2-6, 3-7 and 4-8 of human parathyroid hormone (SEQ ID NO:2). 20. A kit for assaying for whole parathyroid hormone comprising: a) a substantially pure antibody or antibody fragment specific for an initial peptide sequence of whole parathyroid hormone wherein said initial peptide sequence consists of VAL-SER-GLU-ILE-GLN-LEU-MET (SEQ ID NO:3), and wherein at least four amino acids in said initial peptide sequence are part of a reactive portion with said antibody; and b) a labeling component that binds to whole parathyroid hormone, but not to said parathyroid hormone initial peptide sequence VAL-SER-GLU-ILE-GLN-LEU-MET (SEQ ID NO:3). 21. The kit of claim 20, wherein the substantially pure antibody or antibody fragment specifically binds to a parathyroid hormone epitope selected from the group consisting of amino acid residues 2-5, 3-6, 4-7, 5-8, 2-6, 3-7 and 4-8 of human parathyroid hormone (SEQ ID NO:2). 22. A kit for assaying for whole parathyroid hormone comprising: a) a substantially pure antibody or antibody fragment specific for an initial peptide sequence of whole parathyroid hormone wherein said initial peptide sequence consists of VAL-SER-GLU-ILE-GLN-LEU-MET (SEQ ID NO:3), and wherein at least four amino acids in said initial peptide sequence are part of a reactive portion with said antibody; and b) a second antibody bound to a solid support and said second antibody is specific for a portion of whole parathyroid hormone that does not include the domain for adenylate cyclase activation. 23. The kit of claim 22, further comprising a third antibody specific for an epitope left open after the whole parathyroid hormone binds to the first and the second antibodies, thereby forming a precipitating mass. 24. The kit of claim 22, wherein the substantially pure antibody or antibody fragment specifically binds to a parathyroid hormone epitope selected from the group consisting of amino acid residues 2-5, 3-6, 4-7, 5-8, 2-6, 3-7 and 4-8 of human parathyroid hormone (SEQ ID NO:2). 25. A method for measuring an amount of a functional N-terminal parathyroid hormone fragment and whole parathyroid hormone in a sample comprising: a) adding to a sample a first antibody or antibody fragment specific for an initial peptide sequence of whole parathyroid hormone wherein said initial peptide sequence consists of VAL-SER-GLU-ILE-GLN-LEU-MET (SEQ ID NO:3), and wherein at least four amino acids in said initial peptide sequence are part of a reactive portion with said first antibody; b) adding to said sample a second antibody or antibody fragment specific for a peptide comprising amino acid sequence 28 to 34 of human parathyroid hormone (SEQ ID NO:2), which comprises a domain for protein kinase C activation, wherein at least four amino acids in said peptide sequence are a reactive portion with said second antibody; c) allowing said first antibody and second antibody, wherein at least one of which is labeled, to bind to N-terminal parathyroid hormone fragment or whole parathyroid hormone present in said sample, thereby forming a labeled complex; and d) measuring the amount of said labeled complex to measure the amount of said functional N-terminal parathyroid hormone fragment and whole parathyroid hormone in said sample while not detecting an interfering non-(1-84) parathyroid hormone fragment. 26. The method of claim 25, wherein the first antibody or antibody fragment specifically binds to a parathyroid hormone epitope selected from the group consisting of amino acid residues 2-5, 3-6, 4-7, 5-8, 2-6, 3-7 and 4-8 of human parathyroid hormone (SEQ ID NO:2). 27. A method for differentiating between a person having substantially normal parathyroid hormone function and having hyperparathyroidism comprising: a) obtaining a sample from a person to be tested; b) contacting said sample with a substantially pure antibody or antibody fragment specific for an initial peptide sequence of whole parathyroid hormone wherein said initial peptide sequence consists of VAL-SER-GLU-ILE-GLN-LEU-MET (SEQ ID NO:3), and wherein at least four amino acids in said initial peptide sequence are part of a reactive portion with said antibody; and c) assessing binding between said substantially pure antibody or antibody fragment and whole parathyroid hormone, if present in said sample, to measure whole parathyroid hormone level in said person, while not detecting an interfering non-(1-84) parathyroid hormone fragment, and to determine if said person has substantially normal parathyroid hormone function or has hyperparathyroidism. 28. The method of claim 27, wherein the hyperparathyroidism is primary hyperparathyroidism. 29. The method of claim 27, wherein the hyperparathyroidism is secondary hyperparathyroidism. 30. The method of claim 27, wherein the hyperparathyroidism is caused by chronic renal failure. 31. The method of claim 27, wherein the substantially pure antibody or antibody fragment specifically binds to a parathyroid hormone epitope selected from the group consisting of amino acid residues 2-5, 3-6, 4-7, 5-8, 2-6, 3-7 and 4-8 of human parathyroid hormone ID NO:2). 32. The method of claim 27, further comprising comparing the whole parathyroid hormone level determined in said tested person to a whole parathyroid hormone level determined in a normal person without hyperparathyroidism. 33. The method of claim 5, wherein the sample is a rat or human sample. 34. The method of claim 14, wherein the sample is a rat or human sample. 35. The method of claim 18, wherein the sample is a rat or human sample. 36. The method of claim 27, wherein the sample is a rat or human sample.

Details for Patent 6,689,566

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Nps Pharmaceuticals, Inc.	NATPARA	parathyroid hormone	For Injection	125511	01/23/2015		2039-03-29