HYDROXYCHOLESTEROL IMMUNOASSAY

• US 20120115169A1
• Filed: 11/09/2010
• Published: 05/10/2012
• Est. Priority Date: 11/09/2010
• Status: Active Grant

• Current Assignee: Enzo Life Sciences Incorporated (Enzo Biochem Incorporated)
• Original Assignee: Enzo Life Sciences Incorporated (Enzo Biochem Incorporated)
• Inventors: Zipkin, Robert Elliot, Adams, Jeffrey Kroll, Patton, Wayne Forrest, Donegan, James J., Mullenix, Michael C.
• Granted Patent: US 8,969,525 B2
• Time in Patent Office: Days
• US Class Current: 435/7.92
• CPC Class Codes: C07J 43/003 not condensed
• CPC Class Codes: C07J 9/00 Normal steroids containing carbon, hydrogen, halogen or oxygen substituted in position 17 beta by a chain of more than two carbon atoms, e.g. cholane, cholestane, coprostane
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 16/44 against material not provided for elsewhere , e.g. haptens, metals, DNA, RNA, amino acids
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: G01N 2800/2835 Movement disorders, e.g. Parkinson, Huntington, Tourette
• CPC Class Codes: G01N 2800/285 Demyelinating diseases; Multipel sclerosis
• CPC Class Codes: G01N 2800/52 Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
• CPC Class Codes: G01N 33/5308 for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
• CPC Class Codes: G01N 33/56966 Animal cells
• CPC Class Codes: G01N 33/92 involving lipids, e.g. cholesterol , lipoproteins, or their receptors steroid hormones G01N33/743

Abstract

Provided is a derivative of 22-hydroxycholesterol, 24S-hydroxycholesterol, 25-hydroxycholesterol, 26-hydroxycholesterol or 27-hydroxycholesterol. Also provided is a protein conjugated to the above derivative. Further provided is an antibody composition comprising antibodies that specifically bind to 22-hydroxycholesterol, 24S-hydroxycholesterol, 25-hydroxycholesterol, 26-hydroxycholesterol or 27-hydroxycholesterol. Additionally, a method of making antibodies that specifically bind to 22-hydroxycholesterol, 24S-hydroxycholesterol, 25-hydroxycholesterol, 26-hydroxycholesterol or 27-hydroxycholesterol is provided. Also, a method of assaying for 22-hydroxycholesterol, 24S-hydroxycholesterol, 25-hydroxycholesterol, 26-hydroxycholesterol or 27-hydroxycholesterol is provided. Additionally provided is a kit for detecting 22-hydroxycholesterol, 24S-hydroxycholesterol, 25-hydroxycholesterol, 26-hydroxycholesterol or 27-hydroxycholesterol. A method of detecting an enzyme or enzymes utilized in phase II drug metabolism is also provided. Also, a method of detecting an enzyme that synthesizes 22-hydroxycholesterol, 24S-hydroxycholesterol, 25-hydroxycholesterol, 26-hydroxycholesterol or 27-hydroxycholesterol is provided. Further provided is a method of evaluating progression of multiple sclerosis in a patient. Also provided is a method of determining whether a treatment for multiple sclerosis in a patient is effective. Further, a method of evaluating progression of Huntington'"'"'s disease in a patient is provided. Additionally provided is a method of determining whether a treatment for Huntington'"'"'s disease in a patient is effective.

Claims

1. :

A derivative of 22-hydroxycholesterol, 24S-hydroxycholesterol, 25-hydroxycholesterol, 26-hydroxycholesterol or 27-hydroxycholesterol comprising a moiety comprising a free amino, carboxylic acid, or sulfhydryl group covalently attached to the 3-OH of the 22-hydroxycholesterol, 24S-hydroxycholesterol, 25-hydroxycholesterol, 26-hydroxycholesterol or 27-hydroxycholesterol.

3. :

The derivative of claim 1, wherein the derivative is 3-O-succinoyl-24S-hydroxycholesterol

14. :

A protein conjugated to the derivative of claim 1.

2. (canceled)

4. :

A compound comprising 3-O-succinoyl-24S-hydroxycholesterol.

10. :

The compound of claim 4, consisting of

5-9. -9. (canceled)

11-13. -13. (canceled)

15-28. -28. (canceled)

29. :

An antibody composition comprising antibodies that specifically bind to 22-hydroxycholesterol, 24S-hydroxycholesterol, 25-hydroxycholesterol, 26-hydroxycholesterol or 27-hydroxycholesterol.

30. :

The antibody composition of claim 29, wherein the antibodies specifically bind to 24S-hydroxycholesterol and have less than 1% cross-reactivity to cholesterol, 22-hydroxycholesterol, 25-hydroxycholesterol, 26-hydroxycholesterol, 27-hydroxycholesterol and DHEA when compared to the binding of the antibodies to 24S-hydroxycholesterol.

31. :

The antibody composition of claim 30, wherein the antibodies have less than 0.2% cross-reactivity to 22-hydroxycholesterol, cholesterol and DHEA when compared to the binding of the antibodies to 24S-hydroxycholesterol.

32. :

The antibody composition of claim 30, wherein the antibodies have less than 0.01% cross-reactivity to cholesterol when compared to the binding of the antibodies to 24S-hydroxycholesterol.

33. :

The antibody composition of claim 29, wherein the antibodies are polyclonal antibodies.

34. :

The antibody composition of claim 33, wherein the polyclonal antibodies are rabbit antibodies.

35. :

The antibody composition of claim 29, wherein the antibodies are monoclonal antibodies.

36. :

The antibody composition of claim 29, wherein the antibodies are Fab or Fab2 fragments.

37. :

The antibody composition of claim 29, wherein the antibodies are recombinant antibodies.

38-45. -45. (canceled)

46. :

A method of assaying for the hydroxycholesterol 22-hydroxycholesterol, 24S-hydroxycholesterol, 25-hydroxycholesterol, 26-hydroxycholesterol or 27-hydroxycholesterol in a fluid or tissue sample from a mammal, the method comprising combining the sample with antibodies that specifically bind to the hydroxycholesterol (hydroxycholesterol antibodies), then determining whether the antibodies specifically bind to the hydroxycholesterol from the sample, wherein specific antibody binding to the hydroxycholesterol from the sample indicates that the hydroxycholesterol is present in the sample.

51. :

The method of claim 46, wherein the hydroxycholesterol is 24S-hydroxycholesterol and the sample is from a human that is suspected of having cognitive impairment, Huntington'"'"'s disease, Alzheimer'"'"'s disease, or multiple sclerosis.

64. :

The method of claim 46, comprising enzyme linked immunosorbent assay (ELISA).

81. :

A method of detecting an enzyme or enzymes utilized in phase II drug metabolism in a sample, where the enzyme or enzymes are capable of altering 22-hydroxycholesterol, 24S-hydroxycholesterol, 25-hydroxycholesterol, 26-hydroxycholesterol or 27-hydroxycholesterol, the method comprising combining the sample with 22-hydroxycholesterol, 24S-hydroxycholesterol, 25-hydroxycholesterol, 26-hydroxycholesterol or 27-hydroxycholesterol for a time sufficient for the enzyme or enzymes to modify the 22-hydroxycholesterol, 24S-hydroxycholesterol, 25-hydroxycholesterol, 26-hydroxycholesterol or 27-hydroxycholesterol, then assaying the sample using the method of claim 46, wherein a reduction in the amount of the 22-hydroxycholesterol, 24S-hydroxycholesterol, 25-hydroxycholesterol, 26-hydroxycholesterol or 27-hydroxycholesterol detected by the assay indicates that the sample comprises the enzyme or enzymes.

86. :

A method of detecting an enzyme that synthesizes 22-hydroxycholesterol, 24S-hydroxycholesterol, 25-hydroxycholesterol, 26-hydroxycholesterol or 27-hydroxycholesterol in a sample, the method comprising combining the sample with cholesterol for a time sufficient for the enzyme to modify the cholesterol, then assaying the sample using the method of claim 46, wherein an increase in the amount of 22-hydroxycholesterol, 24S-hydroxycholesterol, 25-hydroxycholesterol, 26-hydroxycholesterol or 27-hydroxycholesterol detected by the assay indicates that the sample comprises the enzyme.

47-50. -50. (canceled)

52-63. -63. (canceled)

65-80. -80. (canceled)

82-85. -85. (canceled)

88. (canceled)

90. (canceled)

92. (canceled)