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Last Updated: March 29, 2024

Claims for Patent: 7,820,393


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Summary for Patent: 7,820,393
Title:Methods, kits and antibodies for detecting parathyroid hormone
Abstract: The present invention relates to novel methods and compositions useful for detecting whole parathyroid hormone at a physiological level and parathyroid fragments in a mammalian sample. Such detections may be useful to different parathyroid diseases or disorders in a subject, such as hyperparathyroidism and related bone diseases, from normal or non-disease states. One detects whole or non-fragmented (1 to 84) parathyroid hormone in a biological sample and optionally one or more of a selection of non-whole parathyroid hormone peptide fragments that may or may not function as a parathyroid hormone antagonists. By either comparing values or using independently the value of either the one or more of a selection of non-whole parathyroid hormone peptide fragments, the whole parathyroid hormone, or the combination of these values one is able to differentiate parathyroid and bone related disease states, as well as differentiate such states from normal states.
Inventor(s): Cantor; Thomas L. (El Cajon, CA), Gao; Ping (San Diego, CA)
Assignee: Scantibodies Laboratory, Inc. (Santee, CA)
Application Number:10/617,489
Patent Claims:1. An isolated antibody that specifically binds to an N-terminal sequence of whole parathyroid hormone (PTH) and is capable of detecting said whole PTH at a physiological level in a mammalian sample, with a proviso that said isolated antibody avoids binding to a non-whole PTH fragment.

2. The isolated antibody of claim 1, which is a monoclonal antibody that specifically binds to whole PTH.

3. The isolated antibody of claim 1, which specifically binds to an epitope comprised in PTH.sub.1-5.

4. The isolated antibody of claim 1, which specifically binds to the parathyroid hormone peptide human PTH.sub.1-8, rat PTH.sub.1-8, mouse PTH.sub.1-8, bovine PTH.sub.1-8, canine PTH.sub.1-8, porcine PTH.sub.1-8, horse PTH.sub.1-8, human PTH.sub.1-15, rat PTH.sub.1-15, mouse PTH.sub.1-15, bovine PTH.sub.1-15, canine PTH.sub.1-15, porcine PTH.sub.1-15, or horse PTH.sub.1-15, wherein at least four amino acids in said peptide sequence are part of a reactive portion with the antibody.

5. The isolated antibody of claim 1, which specifically binds to an epitope comprised in PTH.sub.2-5, PTH.sub.2-6, PTH.sub.2-7, PTH.sub.2-8, PTH.sub.2-9, PTH.sub.2-10, PTH.sub.2-11, PTH.sub.2-12, PTH.sub.2-13, PTH.sub.2-14, or PTH.sub.2-15.

6. The isolated antibody of claim 1, wherein the binding between the antibody and the N-terminal sequence of whole PTH is dependent on the presence of amino acid residues 2-5 of the hPTH.

7. The isolated antibody of claim 1, wherein the binding between the antibody and the N-terminal sequence of whole PTH is dependent on the presence of amino acid residues 3-6 of the hPTH.

8. The isolated antibody or antibody fragment of claim 1, wherein the non-whole PTH fragment is a peptide having an amino acid sequence from between PTH.sub.3-84 and PTH.sub.34-84.

9. The isolated antibody of claim 1, wherein the non-whole PTH fragment is a peptide having an amino acid sequence of human PTH.sub.7-84.

10. A method for measuring a physiological level of whole parathyroid hormone (PTH) in a mammalian sample, which method comprises: a) obtaining a sample from a mammal to be tested; b) contacting said sample with an isolated antibody that specifically binds to an N-terminal sequence of whole PTH and is capable of detecting said whole PTH at a physiological level in said mammalian sample, with a proviso that said isolated antibody avoids binding to a non-whole PTH fragment in said sample; and c) assessing a complex formed between said whole PTH, if present in said sample, and said antibody, to measure physiological level of said whole PTH in said mammalian sample, wherein said isolated antibody specifically binds to an epitope comprised in PTH.sub.1-5, PTH.sub.1-6, PTH.sub.1-7, PTH.sub.1-8, PTH.sub.1-9, PTH.sub.1-10, PTH.sub.1-11, PTH-.sub.1-12, PTH.sub.1-13, PTH.sub.1-14 or PTH.sub.1-15.

11. The method of claim 10, wherein the sample is selected from the group consisting of a serum, a plasma and a blood sample.

12. The method of claim 10, wherein the sample, is a clinical sample.

13. The method of claim 10 which is used for clinical management of renal disease subjects, subjects afflicted with osteoporosis or diagnosing primary hyperparathyroidism.

14. The method of claim 10, wherein the mammal is a human.

15. The method of claim 14, wherein the sample is a human clinical sample.

16. The method of claim 10, wherein the antibody is a monoclonal antibody that specifically binds to whole PTH.

17. The method of claim 10, wherein the antibody specifically binds to an epitope comprised in PTH.sub.1-6, PTH.sub.1-8, PTH.sub.1-9, PTH.sub.1-12, or PTH.sub.1-15.

18. The method of claim 10, wherein the antibody specifically binds to the PTH peptide human PTH.sub.1-8, rat PTH.sub.1-8, mouse PTH.sub.1-8, bovine PTH.sub.1-8, canine PTH.sub.1-8, porcine PTH.sub.1-8, horse PTH.sub.1-8, human PTH.sub.1-15, rat PTH.sub.1-15, mouse PTH.sub.1-15, bovine PTH.sub.15, canine PTH.sub.1-15, porcine PTH.sub.1-15, or horse PTH.sub.1-15, wherein at least four amino acids in said peptide sequence are part of a reactive portion with the antibody.

19. The method of claim 10, wherein the non-whole PTH fragment is selected from the group consisting of PTH.sub.3-84, PTH.sub.4-84, PTH.sub.5-84, PTH.sub.6-84, PTH.sub.7-84, PTH.sub.8-84, PTH.sub.9-84, PTH.sub.10-84, PTH.sub.11-84, PTH.sub.12-84, PTH.sub.13-84, PTH.sub.14-84, PTH.sub.15-84, PTH.sub.16-84, PTH.sub.17-84, PTH.sub.18-84, PTH.sub.19-84, PTH.sub.20-84, PTH.sub.21-84, PTH.sub.22-84, PTH.sub.23-84, PTH.sub.24-84, PTH.sub.25-84, PTH.sub.26-84, PTH.sub.27-84, PTH.sub.28-84, PTH.sub.29-84, PTH.sub.30-84, PTH.sub.31-84, PTH.sub.32-84, PTH.sub.33-44 and PTH.sub.34-84.

20. The method of claim 10, wherein the non-whole PTH fragment is a peptide having an amino acid sequence of human PTH.sub.7-84.

21. The method of claim 10, wherein the complex is assessed by a sandwich assay format.

22. The method of claim 21, wherein the antibody that specifically binds to an N-terminal sequence of whole PTH is used as a first antibody in a sandwich format assay, and a second antibody used in the sandwich format assay is an antibody that is capable of binding to a portion of whole PTH other than the N-terminal sequence to which the first antibody binds.

23. The method of claim 22, wherein either the first antibody or the second antibody is attached to a surface and functions as a capture antibody.

24. The method of claim 23, wherein the capture antibody is attached to the surface directly.

25. The method of claim 23, wherein the capture antibody is attached to the surface via a biotin-avidin (or streptavidin) linking pair.

26. The method of claim 10, wherein the complex is assessed by a format selected from the group consisting of an enzyme-linked immunosorbent assay (ELISA), immunoblotting, immunoprecipitation, radioimmunoassay (RIA), immunostaining, latex agglutination, indirect hemagglutination assay (IHA), complement fixation, indirect immunofluorescent assay (IFA), nephelometry, flow cytometry assay, plasmon resonance assay, chemiluminescence assay, lateral flow immunoassay, u-capture assay, inhibition assay and avidity assay.

27. The method of claim 10, wherein the complex is assessed in a homogeneous assay format.

28. The method of claim 10, wherein the physiological level of whole parathyroid hormone PTH is less than 4 pmol/L.

29. The method of claim 10, wherein the physiological level of whole PTH is from about 0.2 pmol/L to about 4 pmol/L.

30. The method of claim 10, which further comprises measuring a PTH peptide fragment level and/or total PTH level.

31. The method of claim 30, wherein said sample is contacted with one or more isolated antibodies, and wherein each of said one or more isolated antibodies specifically bind one or more PTH peptide fragments selected from the group consisting of: PTH.sub.39-84, PTH.sub.1-34, PTH.sub.43-68, PTH.sub.7-84, PTH.sub.39-68, PTH.sub.53-84, PTH.sub.65-84, PTH.sub.44-68, PTH.sub.19-84, PTH.sub.23-84, PTH.sub.1-38, PTH.sub.1-48, PTH.sub.1-58, PTH.sub.1-68, and PTH.sub.1-78.

32. The method of claim 30, which further comprises comparing at least two parameters selected from the group consisting of the whole PTH level, total PTH peptide fragment level, total PTH level, C-terminal PTH fragment (cPTH) level, N-terminal PTH fragment level, and mid-terminal PTH fragment (mPTH) level.

33. The method of claim 32, wherein the results of said comparison are used to determine whether the mammal suffers from a bone turnover related disorder.

34. The method of claim 33, which is used in the diagnosis or monitoring of treatment for adynamic bone disease.

35. The method of claim 32, wherein the comparison is in the form of a ratio or proportion between the whole PTH level and the total PTH level.

36. The method of claim 32, wherein the comparison is in the form of a ratio or proportion between the whole PTH level versus the combined total of the total PTH level minus the whole PTH level.

37. The method of claim 10, which is used for differentiating between a person having substantially normal parathyroid function and having hyperparathyroidism.

38. A kit for measuring a physiological level of whole parathyroid hormone (PTH) in a mammalian sample, which kit comprises, in a container, an isolated antibody that specifically binds to an N-terminal sequence of whole (PTH) and is capable of detecting said whole PTH at a physiological level in a mammalian sample, with a proviso that said isolated antibody avoids binding to a non-(1-84) or non-(1-86) PTH fragment in said sample, wherein said isolated antibody specifically binds to an epitope comprised in PTH.sub.1-5, PTH.sub.1-6, PTH.sub.1-7, PTH.sub.1-8, PTH.sub.1-9, PTH.sub.1-10, PTH.sub.1-11, PTH.sub.1-12, PTH.sub.1-13, PTH.sub.1-14 or PTH.sub.1-15.

39. The method of claim 10, wherein the physiological level of whole PTH is from about 7 picogram/ml to about 39 picogram/ml.

40. The method of claim 23, wherein the capture antibody is attached to the surface indirectly.

41. The method of claim 10, wherein the complex is assessed in a heterogeneous assay format.

42. The method of claim 10, wherein the isolated antibody avoids binding greater than about 90% of a non-whole PTH fragment in said sample.

43. The method of claim 10, wherein the isolated antibody specifically binds to an epitope comprised in PTH.sub.1-5 or PTH.sub.1-6.

44. The method of claim 22, wherein one of the first or the second antibody is labeled.

45. A method for measuring a physiological level of whole parathyroid hormone (PTH) in a mammalian sample, which method comprises: a) obtaining a sample from a mammal to be tested; b) contacting said sample with an isolated antibody that specifically binds to an N-terminal sequence of whole PTH and is capable of detecting said whole PTH at a physiological level in said mammalian sample, with a proviso that said isolated antibody avoids binding to a non-whole PTH fragment; and c) assessing a complex formed between said whole PTH, if present in said sample, and said antibody, to measure physiological level of said whole PTH in said mammalian sample, wherein said antibody specifically binds to an epitope comprised in PTH.sub.1-6, PTH.sub.1-8, PTH.sub.1-9, PTH.sub.1-12, PTH.sub.1-15, or PTH.sub.3-12.

46. The method of claim 45, wherein the non-whole PTH fragment is a peptide having an amino acid sequence of human PTH.sub.7-84.

47. A method for measuring a physiological level of whole parathyroid hormone (PTH) in a mammalian sample, which method comprises: a) obtaining a sample from a mammal to be tested; b) contacting said sample with an isolated antibody that specifically binds to an N-terminal sequence of whole PTH and is capable of detecting said whole PTH at a physiological level in said mammalian sample, with a proviso that said isolated antibody avoids binding to a non-whole PTH fragment; and c) assessing a complex formed between said whole PTH, if present in said sample, and said antibody, to measure physiological level of said whole PTH in said mammalian sample, wherein said antibody specifically binds to the PTH peptide human PTH.sub.1-8, rat PTH.sub.1-8, mouse PTH.sub.1-8, bovine PTH.sub.1-8, canine PTH.sub.1-8, porcine PTH.sub.1-8, horse PTH.sub.1-8, human PTH.sub.1-15, rat PTH.sub.1-15, mouse PTH.sub.1-15, bovine PTH.sub.1-15, canine PTH.sub.1-15, porcine PTH.sub.1-15, or horse PTH.sub.1-15, wherein at least four amino acids in said peptide sequence are part of a reactive portion with the antibody.

48. The method of claim 47, wherein the non-whole PTH fragment is a peptide having an amino acid sequence of human PTH.sub.7-84.

49. The method of claim 10, wherein the isolated antibody that specifically binds to an N-terminal sequence of whole PTH is produced by immunizing a mammal with whole PTH, collecting the antibody from the mammal and isolating the antibody by binding the antibody to an epitope comprised in PTH.sub.1-5, PTH.sub.1-6, PTH.sub.1-7, PTH.sub.1-8, PTH.sub.1-9, PTH.sub.1-10, PTH.sub.1-11, PTH.sub.1-12, PTH.sub.1-13, PTH.sub.1-14 or PTH.sub.1-15, and the isolated antibody specifically binds to whole PTH.

50. The method of claim 49, wherein the mammal is a goat.

51. The method of claim 49, wherein the whole PTH is human whole PTH and the peptide is a human or rat PTH peptide.

52. The method of claim 49, wherein the whole PTH is rat whole PTH and the peptide is a human or rat PTH peptide.

53. The method of claim 10, wherein the mammal is a rat.

54. The method of claim 10, wherein the isolated antibody that specifically binds to an N-terminal sequence of whole PTH is produced by immunizing a mammal with whole PTH, collecting the antibody from the mammal and isolating the antibody using a peptide selected from the group consisting of PTH.sub.1-5, PTH.sub.1-6, PTH.sub.1-7, PTH.sub.1-8, PTH.sub.1-9, PTH.sub.1-10, PTH.sub.1-11, PTH.sub.1-12, PTH.sub.1-13, PTH.sub.1-14 and PTH.sub.1-15, and the isolated antibody specifically binds to whole PTH.

55. The method of claim 32, wherein the results of said comparison are used to monitor bone disease or disorder related treatment.

56. The isolated antibody or antibody fragment of claim 8, wherein the non-whole PTH fragment is selected from the group consisting of PTH.sub.4-84, PTH.sub.5-84, PTH.sub.6-84, PTH.sub.7-84, PTH.sub.8-84, PTH.sub.9-84, PTH.sub.10-84, PTH.sub.11-84, PTH.sub.12-84, PTH.sub.13-84, PTH.sub.14-84, and PTH.sub.15-84.

57. The method of claim 19, wherein the non-whole PTH fragment is selected from the group consisting of PTH.sub.4-84, PTH.sub.5-84, PTH.sub.6-84, PTH.sub.7-84, PTH.sub.8-84, PTH.sub.9-84, PTH.sub.10-84, PTH.sub.11-84, PTH.sub.12-84, PTH.sub.13-84, PTH.sub.14-84 and PTH.sub.15-84.

58. The isolated antibody of claim 1, which is a polyclonal antibody that specifically binds to whole PTH.

59. The isolated antibody of claim 1, which is an antibody fragment that specifically binds to whole PTH.

60. The isolated antibody of claim 1, which specifically binds to an epitope comprised in PTH.sub.1-6.

61. The isolated antibody of claim 1, which specifically binds to an epitope comprised in PTH.sub.1-7.

62. The isolated antibody of claim 1, which specifically binds to an epitope comprised in PTH.sub.1-8.

63. The isolated antibody of claim 1, which specifically binds to an epitope comprised in PTH.sub.1-9.

64. The isolated antibody of claim 1, which specifically binds to an epitope comprised in PTH.sub.1-10.

65. The isolated antibody of claim 1, which specifically binds to an epitope comprised in PTH.sub.1-11.

66. The isolated antibody of claim 1, which specifically binds to an epitope comprised in PTH.sub.1-12.

67. The isolated antibody of claim 1, which specifically binds to an epitope comprised in PTH.sub.1-13.

68. The isolated antibody of claim 1, which specifically binds to an epitope comprised in PTH.sub.1-14.

69. The isolated antibody of claim 1, which specifically binds to an epitope comprised in PTH.sub.1-15.

70. The isolated antibody of claim 1, which specifically binds to an epitope comprised in PTH.sub.3-12.

71. The isolated antibody of claim 1, which specifically binds to the parathyroid hormone peptide human PTH.sub.1-8, wherein at least four amino acids in said peptide sequence are part of a reactive portion with the antibody.

72. The isolated antibody of claim 1, which specifically binds to the parathyroid hormone peptide rat PTH.sub.1-8, wherein at least four amino acids in said peptide sequence are part of a reactive portion with the antibody.

73. The isolated antibody of claim 1, which specifically binds to the parathyroid hormone peptide human PTH.sub.1-12, wherein at least four amino acids in said peptide sequence are part of a reactive portion with the antibody.

74. The isolated antibody of claim 1, which specifically binds to the parathyroid hormone peptide rat PTH.sub.1-12, wherein at least four amino acids in said peptide sequence are part of a reactive portion with the antibody.

75. The isolated antibody of claim 1, wherein the isolated antibody avoids binding greater than about 90% of a non-whole PTH fragment in the mammalian sample.

76. The isolated antibody of claim 1, wherein the isolated antibody avoids binding greater than about 95% of a non-whole PTH fragment in the mammalian sample.

77. The isolated antibody of claim 1, wherein the isolated antibody avoids binding greater than about 99% or more of a non-whole PTH fragment in the mammalian sample.

78. The method of claim 10, wherein the antibody is a polyclonal antibody that specifically binds to whole PTH.

79. The method of claim 10, wherein the antibody is an antibody fragment that specifically binds to whole PTH.

80. The method of claim 10, wherein the complex is assessed by a competitive assay format.

81. The method of claim 33, which is used in the diagnosis or monitoring of treatment for severe hyperparathyroidism.

82. The method of claim 10, which is used for monitoring parathyroid related bone disease and treatment.

83. The method of claim 10, which is used for monitoring effects of therapeutic treatment for hyperparathyroidism.

84. The method of claim 10, which is used for diagnosing parathyroid related bone disease.

85. The method of claim 10, wherein the isolated antibody specifically binds to an epitope comprised in PTH.sub.1-6.

86. The method of claim 10, wherein the isolated antibody specifically binds to an epitope comprised in PTH.sub.1-7.

87. The method of claim 10, wherein the isolated antibody specifically binds to an epitope comprised in PTH.sub.1-8.

88. The method of claim 10, wherein the isolated antibody specifically binds to an epitope comprised in PTH.sub.1-9.

89. The method of claim 10, wherein the isolated antibody specifically binds to an epitope comprised in PTH.sub.1-10.

90. The method of claim 10, wherein the isolated antibody specifically binds to an epitope comprised in PTH.sub.1-11.

91. The method of claim 10, wherein the isolated antibody specifically binds to an epitope comprised in PTH.sub.1-12.

92. The method of claim 10, wherein the isolated antibody specifically binds to an epitope comprised in PTH.sub.1-13.

93. The method of claim 10, wherein the isolated antibody specifically binds to an epitope comprised in PTH.sub.1-14.

94. The method of claim 10, wherein the isolated antibody specifically binds to an epitope comprised in PTH.sub.1-15.

95. The method of claim 10, wherein the isolated antibody avoids binding greater than about 95% of a non-whole PTH fragment in the mammalian sample.

96. The method of claim 10, wherein the isolated antibody avoids binding greater than about 99% or more of a non-whole PTH fragment in the mammalian sample.

97. The isolated antibody of claim 1, which specifically binds to at least four amino acids in PTH.sub.1-8 sequence.

98. The isolated antibody of claim 97, which avoids binding to PTH.sub.7-84 fragment.

99. The isolated antibody of claim 1, which specifically binds to PTH.sub.1-6 sequence as part of whole PTH.

100. The isolated antibody of claim 1, which specifically binds to PTH.sub.1-7 sequence as part of whole PTH.

101. The isolated antibody of claim 1, which specifically binds to PTH.sub.1-8 sequence as part of whole PTH.

102. The isolated antibody of claim 1, which specifically binds to PTH.sub.1-9 sequence as part of whole PTH.

103. The isolated antibody of claim 1, which specifically binds to PTH.sub.1-10 sequence as part of whole PTH.

104. The isolated antibody of claim 1, which specifically binds to PTH.sub.1-11 sequence as part of whole PTH.

105. The isolated antibody of claim 1, which specifically binds to PTH.sub.1-12 sequence as part of whole PTH.

106. The isolated antibody of claim 1, which specifically binds to PTH.sub.1-13 sequence as part of whole PTH.

107. The isolated antibody of claim 1, which specifically binds to PTH.sub.1-14 sequence as part of whole PTH.

108. The isolated antibody of claim 1, which specifically binds to PTH.sub.1-15 sequence as part of whole PTH.

109. A method for measuring a physiological level of whole parathyroid hormone (PTH) in a mammalian sample, which method comprises: a) obtaining a sample from a mammal to be tested; b) contacting said sample with an isolated antibody that specifically binds to an N-terminal sequence of whole PTH and is capable of detecting said whole PTH at a physiological level in said mammalian sample, with a proviso that said isolated antibody avoids binding to a non-whole PTH fragment in said sample; and c) assessing a complex formed between said whole PTH, if present in said sample, and said antibody, to measure physiological level of said whole PTH in said mammalian sample, wherein said isolated antibody specifically binds to PTH.sub.1-6 sequence as part of whole PTH, PTH.sub.1-7 sequence as part of whole PTH, PTH.sub.1-8 sequence as part of whole PTH, PTH.sub.1-9 sequence as part of whole PTH, PTH.sub.1-10 sequence as part of whole PTH, PTH.sub.1-11 sequence as part of whole PTH, PTH.sub.1-12 sequence as part of whole PTH, PTH.sub.1-13 sequence as part of whole PTH, PTH.sub.1-14 sequence as part of whole PTH, or PTH.sub.1-15 sequence as part of whole PTH.

110. The method of claim 109, wherein the isolated antibody avoids binding to PTH.sub.7-84 fragment.

111. The method of claim 109, wherein the isolated antibody specifically binds to PTH.sub.1-6 sequence as part of whole PTH.

112. The method of claim 109, wherein the isolated antibody specifically binds to PTH.sub.1-7 sequence as part of whole PTH.

113. The method of claim 109, wherein the isolated antibody specifically binds to PTH.sub.1-8 sequence as part of whole PTH.

114. The method of claim 109, wherein the isolated antibody specifically binds to PTH.sub.1-9 sequence as part of whole PTH.

115. The method of claim 109, wherein the isolated antibody specifically binds to PTH.sub.1-10 sequence as part of whole PTH.

116. The method of claim 109, wherein the isolated antibody specifically binds to PTH.sub.1-11 sequence as part of whole PTH.

117. The method of claim 109, wherein the isolated antibody specifically binds to PTH.sub.1-12 sequence as part of whole PTH.

118. The method of claim 109, wherein the isolated antibody specifically binds to PTH.sub.1-13 sequence as part of whole PTH.

119. The method of claim 109, wherein the isolated antibody specifically binds to PTH.sub.1-14 sequence as part of whole PTH.

120. The method of claim 109, wherein the isolated antibody specifically binds to PTH.sub.1-15 sequence as part of whole PTH.

121. A kit for measuring a physiological level of whole parathyroid hormone (PTH) in a mammalian sample, which kit comprises, in a container, an isolated antibody that specifically binds to an N-terminal sequence of whole PTH and is capable of detecting said whole PTH at a physiological level in a mammalian sample, with a proviso that said isolated antibody avoids binding to a non-whole PTH fragment in said sample, wherein said isolated antibody specifically binds to PTH.sub.1-6 sequence as part of whole PTH, PTH.sub.1-7 sequence as part of whole PTH, PTH.sub.1-8 sequence as part of whole PTH, PTH.sub.1-9 sequence as part of whole PTH, PTH.sub.1-10 sequence as part of whole PTH, PTH.sub.1-11 sequence as part of whole PTH, PTH.sub.1-12 sequence as part of whole PTH, PTH.sub.1-13 sequence as part of whole PTH, PTH.sub.1-14 sequence as part of whole PTH, or PTH.sub.1-15 sequence as part of whole PTH.

122. The kit of claim 121, wherein the isolated antibody avoids binding to PTH.sub.7-84 fragment.

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