Antibodies specific to deoxythymidine 5'-phosphate

Antibodies to deoxythymidine 5'-phosphate (dpT) were elicited in rabbits using bovine serum albumin conjugate of the nucleotide. To det. the specificity of the antibodies, binding of tritiated hapten, [3H]dpT, to the g-globulins prepared from the antisera and inhibition of this binding by nonradioactive dpT and related compds. was measured by nitrocellulose filter assay. The results showed that the antibodies (i) were specific for the whole molecule of dpT, (ii) had very high steric requirement for recognizing the hapten, (iii) showed negligible cross reactivity with other mononucleotides, and (iv) recognized dpT residue in an oligonucleotide but showed preference for the dpT at the 3' end over the one at the 5'-end of the oligonucleotide. The property (iii) was used in developing a sensitive radioimmunoassay for the quantitation of dpT in the presence of all the 4 normal mononucleotides. The relability of this method was demonstrated by quantitating the amts. of dpT in oligonucleotides and DN samples of known base component