Santella R M, Dharmaraja N, Gasparro F P, Edelson R L
Nucleic Acids Res. 1985 Apr 11;13(7):2533-44. doi: 10.1093/nar/13.7.2533.
A panel of monoclonal antibodies have been developed which specifically recognize DNA modified by 8-methoxypsoralen (8-MOP) and ultraviolet A light (320-400 nm) (UVA). These antibodies have been characterized as to sensitivity and specificity by an enzyme linked immunosorbent assay (ELISA). In a competitive ELISA with the most sensitive antibody, 50% inhibition of antibody binding occurred at 17 fmole 8-MOP-DNA photo adducts. One adduct per 10(7) bases could be reliably detected. There was also some antibody cross-reactivity with DNAs modified by 4' aminomethyl-4, 5, 8-trimethylpsoralen and 4', 5-dimethylangelicin as well as DNA isolated from cells treated with 8-MOP and UVA. The primary specificity of one of the antibodies was shown to be the 4', 5' thymine monoadduct by competitive inhibition studies using HPLC fractions of an enzymatic digest of 8-MOP poly(dA-dT) . poly(dA-dT). These antibodies should allow the quantitation of adduct levels in various in vitro systems as well as humans exposed clinically to 8-MOP and UVA.
已研发出一组单克隆抗体,它们能特异性识别经8-甲氧基补骨脂素(8-MOP)和紫外线A光(320 - 400纳米)(UVA)修饰的DNA。这些抗体通过酶联免疫吸附测定(ELISA)对敏感性和特异性进行了表征。在使用最敏感抗体的竞争性ELISA中,当8-MOP - DNA光加合物达到17飞摩尔时,抗体结合受到50%的抑制。每10⁷个碱基中的一个加合物能够被可靠地检测到。这些抗体还与经4'-氨甲基-4,5,8-三甲基补骨脂素和4',5-二甲基白芷素修饰的DNA以及从用8-MOP和UVA处理的细胞中分离出的DNA存在一定的交叉反应。通过使用8-MOP聚(dA - dT)·聚(dA - dT)酶解产物的HPLC馏分进行竞争性抑制研究,表明其中一种抗体的主要特异性是4',5'-胸腺嘧啶单加合物。这些抗体应能对各种体外系统以及临床暴露于8-MOP和UVA的人体中的加合物水平进行定量。