Roberts J C, Figard S D, Mercer-Smith J A, Svitra Z V, Anderson W L, Lavallee D K
Medical Radioisotopes Research Program, Los Alamos National Laboratory, NM 87545.
J Immunol Methods. 1987 Dec 24;105(2):153-64. doi: 10.1016/0022-1759(87)90261-4.
Methods were developed to label antibodies with copper-67, a potentially useful medical radioisotope, using the porphyrin chelating agent N-benzyl-5,10,15,20-tetrakis(4-carboxyphenyl) porphine. The porphyrin was activated for coupling using either (1) 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide HCl and N-hydroxysuccinimide or (2) 1,1'-carbonyldiimidazole. The coupling reactions were optimized as a function of activation time, coupling time, coupling pH, and reagent concentrations to achieve maximum coupling to IgG monomer. Sodium dodecylsulfate polyacrylamide gel electrophoresis was used to determine coupling yields. After purification by gel filtration, the antibody-porphyrin conjugates were labeled with copper-67 in aqueous solution. The coupling protocols were used to label antibodies from several species, demonstrating the general utility of these methods. Characterization of the conjugates indicated that the porphyrin label was attached randomly to the IgG molecule. Antigen binding capacities after conjugation were unaltered or slightly lowered as determined by a competitive ELISA.
研究人员开发了一些方法，使用卟啉螯合剂N-苄基-5,10,15,20-四（4-羧基苯基）卟啉，用潜在有用的医用放射性同位素铜-67标记抗体。使用（1）1-乙基-3-（3-二甲基氨基丙基）碳二亚胺盐酸盐和N-羟基琥珀酰亚胺或（2）1,1'-羰基二咪唑将卟啉活化以进行偶联。根据活化时间、偶联时间、偶联pH值和试剂浓度优化偶联反应，以实现与IgG单体的最大偶联。使用十二烷基硫酸钠聚丙烯酰胺凝胶电泳来确定偶联产率。通过凝胶过滤纯化后，抗体-卟啉缀合物在水溶液中用铜-67标记。这些偶联方案用于标记来自多个物种的抗体，证明了这些方法的通用性。缀合物的表征表明，卟啉标记随机附着在IgG分子上。通过竞争性ELISA测定，偶联后抗原结合能力未改变或略有降低。
