Antibodies to 25-hydroxyvitamin d2 and d3 and uses thereof

Abstract

Provided herein are antigenic molecules that can be used to generate antibodies capable of binding to a vitamin D derivative, such as 25-hydroxyvitamin D2 and/or 25-hydroxyvitamin D3, or a 25-hydroxyvitamin D analog, such as a vitamin D-C22 immunogenic molecule or compound. Antibodies produced using these antigenic molecules, and related antigenic compounds, are also described. In addition, disclosed herein are methods for detecting vitamin D deficiency in a subject, methods for treating a subject suspected of having a vitamin D deficiency, methods for monitoring progression of vitamin D deficiency in a subject, and methods for monitoring treatment of vitamin D deficiency in a subject in need thereof. The methods involve the detection or quantification of 25-hydroxyvitamin D2 and D3. Also provided are methods and reagents for the detection or quantification of 25-hydroxyvitamin D2 and D3, methods for stabilizing vitamin D analogs, and methods for separating 25-hydroxyvitamin D2 and D3 from vitamin D binding protein in a biological sample.

What is claimed: 
     
         1 . A method for treating a subject suspected of having a vitamin D deficiency, said method comprising:
 determining the level of total 25-hydroxyvitamin D in a biological sample derived from said subject;   and if a decrease between the level in the biological sample relative to the level in a normal control or a threshold level of 30 ng/mL is determined, administering to said subject a treatment for vitamin D deficiency.   
     
     
         2 . A method for monitoring treatment of vitamin D deficiency in a subject in need thereof, said method comprising:
 determining the level of total 25-hydroxyvitamin D in a first biological sample derived from said subject at a first time; and   determining the level of total 25-hydroxyvitamin D in a second biological sample derived from said subject at a second time later than said first time and following treatment of said subject for said vitamin D deficiency;   wherein an increase in or stabilization of the level in the second biological sample relative to the level in the first biological sample is indicative of efficacy of the treatment of the vitamin D deficiency in said subject, and   wherein a decrease in the level in the second biological sample relative to the level in the first biological sample is indicative of inefficacy of the treatment of the vitamin D deficiency in said subject.   
     
     
         3 . The method of  claim 1  or  2 , wherein the level of 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3 is determined by contacting said biological sample with an antibody that recognizes both 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3 or an antigen-binding fragment thereof. 
     
     
         4 . A hybridoma cell that produces a monoclonal antibody that binds 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3. 
     
     
         5 . A method for detecting or determining the level of total 25-hydroxyvitamin D in a biological sample comprising contacting said sample with an antibody, or antigen-binding fragment thereof, that binds 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3. 
     
     
         6 . The method of  claim 5  wherein said determining step comprises a competitive immunoassay using fluorescein conjugated vitamin D-C22. 
     
     
         7 . The method of  claim 5  wherein said antibodies and antigen-binding fragments are used to detect a vitamin D derivative, such as 25-hydroxyvitamin D2 and/or 25-hydroxyvitamin D3. 
     
     
         8 . A kit comprising an antibody, or an antigen-binding fragment thereof, that binds 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3 in a sample. 
     
     
         9 . A diagnostic device comprising an antibody, or an antigen-binding fragment thereof, that binds 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3 in a sample. 
     
     
         10 . The kit of  claim 8  further comprising an alkaline phosphatase-conjugated vitamin D-C22-bovine serum albumin (BSA). 
     
     
         11 . The diagnostic device of  claim 9  further comprising an alkaline phosphatase conjugated vitamin D-C22-bovine serum albumin (BSA). 
     
     
         12 . The device of  claim 9  wherein said device performs enhanced chemiluminescence (ECL), enzyme immunoassay (EIA), immunohistochemistry (IHC), western blot analysis, radioimmunoassay (RIA), immunofluorescence, equilibrium dialysis, immunodifferentiation, or enzyme-linked immunosorbant assay (ELISA).