摘要
A photoreactive azido analog of the trypanocide ethidium bromide, 3-amino-8-azido-5-ethyl-6-phenylphenanthridinium chloride, attached covalently to calf thymus DNA (CT DNA) by photoaffinity labeling, was used to generate antibodies for the drug analog. The specificity of the antiserum was tested using enzyme-linked immunoadsorbant assays (ELISA) against immobilized antigen (photoaffinity labeled DNA) and by both the avidin-biotin peroxidase reaction and indirect immunofluorescence performed on smears of drug treated trypanosomes. The reaction of the antiserum with the covalently bound drug adduct was diminished effectively by prior incubation with an excess of ethidium monoazide, ethidium diazide, and ethidium bromide, and to a lesser extent by the DNA-ethidium complex, the diazide-DNA or RNA adduct, and the monoazide-RNA adduct. DNA which had been photoaffinity labeled with either the propidium or the acridine moiety did not react. The antiserum recognition of DNA photoaffinity labeled with ethidium monoazide was based on the substituted phenanthridinium ring system of the parent ethidium, as evidenced by competition binding studies involving the free monoazido analog (EA1), the diazido analog (EA2), and the parent compound, ethidium bromide (EB). This approach and the sensitivity it provides should prove useful for identifying the distribution and fate of covalently bound drugs resulting from antiparasitic drug treatment, and for studying their roles in antiparasitic action.
摘要译文
锥虫溴化乙锭，3-氨基-8-叠氮基-5-乙基-6-苯基菲啶氯化物的光反应性叠氮基类似物，通过光亲和标记与小牛胸腺DNA（CT DNA）共价连接，用于产生药物类似物的抗体。ssays（ELISA）针对固定化抗原（光亲和标记的DNA）和抗生物素蛋白 - 生物素过氧化物酶反应和间接免疫荧光对药物处理的锥虫的涂片进行。通过预先与过量的单叠氮乙锭，二叠氮化乙锭和溴化乙锭一起温育，有效地减少了抗血清与共价结合的药物加合物的反应，并且在较小程度上由DNA-乙锭复合物，二叠氮化物-DNA或RNA加合物和单叠氮-RNA加合物组成。已经用丙锭或吖啶部分标记的光亲和性的DNA没有反应。用单乙锭标记的DNA光亲和性的抗血清识别基于亲本乙锭的取代的菲啶鎓环系，如通过竞争结合研究所证明的，其涉及游离的单叠氮基类似物（EA1），二叠氮基类似物（EA2）和母体化合物溴化乙锭（EB）。这种方法及其提供的灵敏度应该有助于确定抗寄生虫药物治疗产生的共价结合药物的分布和命运，并研究他们在抗寄生虫作用中的作用。
Paul E.Omholt;Betty A.Cox;Laura C.Prine;SuzanneByrd;Lerena W.Yielding;K.LemoneYielding;. Use of drug-specific antibodies to identify ethidium adducts produced in Trypanosoma brucei by photoaffinity labeling[J]. Acta Tropica, 1993,55(4): 191-204