Hydroxycholesterol immunoassay

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.

What is claimed: 
     
         1 . A method of making antibodies that specifically bind to hydroxycholesterol, 22-hydroxycholesterol, 24S-hydroxycholesterol, 25-hydroxycholesterol, 26-hydroxycholesterol or 27-hydroxycholesterol, the method comprising
 a. preparing an immunogen by conjugating a hydroxycholesterol selected from hydroxycholesterol, 22-hydroxycholesterol, 24S-hydroxycholesterol, 25-hydroxycholesterol, 26-hydroxycholesterol and 27-hydroxycholesterol or a derivative of hydroxycholesterol, 22-hydroxycholesterol, 24S-hydroxycholesterol, 25-hydroxycholesterol, 26-hydroxycholesterol and 27-hydroxycholesterol to a carrier protein;   b. immunizing a vertebrate with the immunogen under conditions such that the immune system of the vertebrate makes antibodies to the hydroxycholesterol; and   c. collecting the antibodies to the hydroxycholesterol from the vertebrate.   
     
     
         2 . The method of  claim 1 , wherein the hydroxycholesterol is an O-succinoyl derivative. 
     
     
         3 . The method of  claim 2 , wherein the hydroxycholesterol derivative is 3-O-succinoyl-24S-hydroxycholesterol 
       
         
           
           
               
               
           
         
       
     
     
         4 . The method of  claim 1 , wherein the carrier protein is keyhole limpet hemocyanin, bovine serum albumin or ovalbumin. 
     
     
         5 . The method of  claim 1 , wherein the vertebrate is a rabbit, a goat, a mouse, a chicken, or a sheep. 
     
     
         6 . The method of  claim 1 , wherein the vertebrate is a rabbit. 
     
     
         7 . The method of  claim 1 , wherein the vertebrate is a mouse. 
     
     
         8 . The method of  claim 1 , wherein the antibodies collected from the vertebrate are polyclonal antibodies. 
     
     
         9 . The method of  claim 1 , wherein the antibodies collected from the vertebrate are monoclonal antibodies.