N-琥珀酰亚胺-3-碘[^(125)I]苯甲酸酯标记人IgG  

Indirect Radioiodination of Human IgG With N-Succinimidyl-3-Iodo[^(125)I] Benzoate

作　　者：刘振锋[1] 汪勇先[2] 董墨[2] 周伟[2] 夏姣云[2] 尹端沚[2] 李林法[1] 

机构地区：[1]浙江大学医学院附属第一医院核医学科,杭州310003 [2]中国科学院上海应用物理研究所,上海201800

出　　处：《核化学与放射化学》2007年第3期171-177,共7页Journal of Nuclear and Radiochemistry

基　　金：中国科学院创新工程重大项目(KJCXI-SW-08);国家自然科学基金资助项目(10405034);上海市科委纳米专项资助项目(0352nm120)

摘　　要：研究能有效降低抗体的体内脱碘的标记方法。碘标记N-琥珀酰亚胺-3-(三正丁基锡)苯甲酸酯(ATE)前体,得到N-琥珀酰亚胺-3-碘[125I]苯甲酸酯(S125IB),分别与人IgG和抗人肝癌单抗(Hepama-1)进行偶联,探索最佳标记条件,并测定标记物的稳定性和生物活性,研究直接标记和间接标记的Hepama-1在正常小鼠体内的生物学分布。结果表明,用N-氯代琥珀酰亚胺(NCS)法125I标记ATE前体,在ATE用量为25～100μg、NCS用量为10～20μg、磷酸盐缓冲溶液(PBS)用量为10～20μL、反应时间为5 min时,标记率大于95%;S125IB和人IgG的偶联率最高可达75%,偶联产物稳定性、生物活性良好;与Hepama-1偶联率可达75%以上。生物分布的对比实验证明,1,3,4,6-四氯-3α,6α-二苯甘脲(Iodogen)直接标记的Hepama-1在甲状腺的放射性摄取率(脱碘显示)最高是S125IB间接标记的Hepama-1的87.9倍。这说明以ATE为前体的放射性碘间接标记蛋白质方法与传统的碘直接标记方法相比较,在解决体内严重脱碘问题上具有明显的优越性。The objective of this study was to develop an acylation method for the radioiodination of monoclonal antibodies that could decrease the loss of radioiodine in vivo. Preparation of N- succinimidyl-3-iodobenzoate（S^125 IB） from the organoth precursor, N-succinimidyl-3-（tri-n-butylstannyl）benzoate（ATE） proceeds in more than 95 % labelling yield, when the mass of ATE and NCS are respectively 25-100 μg and 10-20 μg, and the volume of PBS is 10-20 μL, and reaction time is 5 min. IgG is labeled using S^125IB in up to 75% conjugation efficiency and with well retained immunoreactivity to sheep anti-human IgG. Hepama-1 is also labeled using S^125 IB in more than 75% conjugation efficiency. Paired-label biodistribution studies in normal mice demonstrate that thyroid uptake（a monitor of dehalogenation） of Hepama-1 labeled by S^125 IB method is up to 87.9 times lower than that of Hepama-1 labeled with Iodogen. This result suggests that S^125IB offers significant advantages for labeling proteins, antibodies over other conventional methods for protein radioiodination

关 键 词：碘标记 人IGG N-琥珀酰亚胺-3-(三正丁基锡)苯甲酸酯(ATE) 

分 类 号：R142.3[医药卫生—公共卫生与预防医学]