Production and specificity of antisera raised against 25-hydroxyvitamin D3-[C-3]-bovine serum albumin conjugates

Article abstract of DOI:10.1016/0039-128X(92)90043-9

In order to obtain specific antisera for use in the enzyme immunoassay of 25-hydroxyvitamin D₃, three hapten-carrier conjugates having different lengths of bridges at the C-3 position were prepared from 25-hydroxyvitamin D₃ by coupling with bovine serum albumin using the active ester method. The specificity of anti-25-hydroxyvitamin D₃ antisera elicited in rabbits was tested by a cross-reaction study with closely related secosterols and by measuring the plasma levels of 25-hydroxyvitamin D₃ by means of radioimmunoassay using tritium-labeled antigen. The results indicated that the specificity of the antisera obtained is higher than that of vitamin D-binding protein, and that some of these antisera are suitable for enzyme immunoassay.