Arif Z, Ali R
Department of Biochemistry, Faculty of Medicine, Aligarh Muslim University, India.
Arch Biochem Biophys. 1996 May 15;329(2):191-8. doi: 10.1006/abbi.1996.0208.
We report the formation of photocrosslink (diad-duct) between duplex poly(dA-dT) and 8-methoxypsoralen inferred on the basis of UV and fluorescence characteristics, Tm, nuclease S1 digestibility, and hydroxyapatite column chromatography. The photocrosslink was highly immunogenic inducing high titer antibodies in goat. The affinity constant of antigen-antibody interaction quantitated by precipitin titration was found to be 2.71 x 10(9) M-1, calculated by Langmuir isotherm plot. These antibodies were highly specific showing recognition of almost exclusively of nucleic acid-furocoumarin photoadducts and could be used as a probe to detect and quantitate DNA modified with furocoumarins in health and disease. Antibodies against poly(dA-dT)-furocoumarin photocrosslink might be a better probe because of repeated A-T sequences on the immunogen.
我们报道了基于紫外和荧光特性、熔点(Tm)、核酸酶S1消化率以及羟基磷灰石柱色谱法推断出的双链聚(dA-dT)与8-甲氧基补骨脂素之间光交联(二元加合物)的形成。这种光交联具有高度免疫原性,能在山羊体内诱导产生高滴度抗体。通过沉淀素滴定法定量的抗原-抗体相互作用亲和常数,根据朗缪尔等温线图计算得出为2.71×10⁹ M⁻¹。这些抗体具有高度特异性,几乎只识别核酸-呋喃香豆素光加合物,可作为一种探针用于检测和定量健康及疾病状态下用呋喃香豆素修饰的DNA。由于免疫原上存在重复的A-T序列,抗聚(dA-dT)-呋喃香豆素光交联的抗体可能是一种更好的探针。