Grishchenko A M, Tereshchenko M I, Potopal'skiĭ A I, Voloshchuk T P
Mol Biol (Mosk). 1987 May-Jun;21(3):814-9.
DNA binding activity of rabbit antiserum against calf spleen DNA's modified by thiophosphamide (DNA-T) was studied by means of solid enzyme immunoassays (ELISA). The studies demonstrated the preferential binding of the immobilized DNA-T compared to immobilized single-stranded DNA (ss-DNA) and only small preference compared to native DNA. Two antisera against DNA-T were purified by affinity chromatography on a ss-DNA-CNBr agarose from antibodies to calf spleen ss-DNA. They interacted only with the immobilized DNA-T, but not with ss-DNA or native DNA. These results demonstrated that DNA modification by thiophosphamide, decreases the immunogenicity of usual nitrogen-containing DNA bases, but detected new immunogenic specificity for adducts. Detection of new immunogenic specificity in DNA's alkylated by thiophosphamide, resulted in the development of a sensitive enzyme immunoassay for the detection of these adducts in nucleic acids, in monitoring their formation, persistence and repair damages in DNA.
采用固相酶免疫测定法（ELISA）研究了兔抗硫代磷酰胺修饰的小牛脾DNA（DNA-T）抗血清的DNA结合活性。研究表明，与固定化单链DNA（ss-DNA）相比，固定化DNA-T具有优先结合性，而与天然DNA相比，优先性较小。通过在ss-DNA-CNBr琼脂糖上进行亲和层析，从小牛脾ss-DNA抗体中纯化出两种抗DNA-T抗血清。它们仅与固定化DNA-T相互作用，而不与ss-DNA或天然DNA相互作用。这些结果表明，硫代磷酰胺对DNA的修饰降低了常见含氮DNA碱基的免疫原性，但检测到加合物具有新的免疫原性特异性。检测硫代磷酰胺烷基化DNA中的新免疫原性特异性，导致开发出一种灵敏的酶免疫测定法，用于检测核酸中的这些加合物，监测它们在DNA中的形成、持久性和修复损伤情况。
