用于凋亡显像的Cys-Annexin V冻干品的制备和质量控制  

Preparation and quality control of freeze-dried products of Cys-Annexin V for apoptosis imaging

作　　者：蒋泉福[1] 陆春雄[1] 胡敏进[2] 俞惠新[1] 华子春[2,3] 

机构地区：[1]江苏省原子医学研究所卫生部核医学重点实验室江苏省分子核医学重点实验室,无锡214063 [2]江苏靶标生物医药研究所有限公司常州南京大学高新技术研究院,常州213164 [3]南京大学医药生物技术国家重点实验室,南京210093

出　　处：《中国新药杂志》2015年第21期2507-2510,共4页Chinese Journal of New Drugs

基　　金：卫生部课题"一种新型99mTc标记膜联蛋白V变体用于细胞凋亡显像的临床前研究"(W201207);江苏省科技基础设施建设计划--科技公共服务平台(BM2012066);南京大学医药生物技术国家重点实验室开放基金(KF-GN-201304);江苏省自然基金(BK20151119);江苏省卫生科技计划项目(Q201204)

摘　　要：目的：制备Cys-Annexin V冻干品,并建立相应的质量控制方法。方法：将一定量的新型膜联蛋白变体蛋白（Cys-Annexin V）、氯化亚锡和葡庚糖酸钠溶于磷酸缓冲液,制备成冻干品;使用Bradford法测冻干品中蛋白含量;HPLC测定标记率;电位滴定法测冻干品中氯化亚锡含量;并对该冻干品的稳定性进行考察。结果：制备得到白色粉末冻干品,该冻干品的99mTc标记率大于90%,Bradford法测定蛋白含量,在0.01~0.1 mg·m L-1范围内线性良好,平均回收率为96.6%,RSD为6.25%。该冻干品对高温敏感,但在加速实验下6个月和长期稳定性实验下12个月是稳定的。结论：制备工艺简单易行,稳定性好,质量可控。Objective： To prepare freeze-dried kit of Cys-Annexin V and establish the quality control method. Methods： Freeze-dried kit of Cys-Annexin V were prepared with Cys-Annexin V,stannous chloride dihydrate and glucoheptonate. Content of Cys-Annexin V in the freeze-dried kit was determined by the Bradford＇s dye-binding method. The labeling yield of Cys-Annexin V was determined by HPLC. The content of stannous chloride in freezedried kit was determined by potentiometric titration. The stability test was also performed. Results： Homogenous pellet was obtained as white powder and the labeling yield of the kit was above 90% with99 mTc. A good linear relationship was obtained in the range of 0. 01 ~ 0. 1 mg·m L- 1,the average recovery of sample was 96. 6%,and RSD was 6. 25%. The kit was sensitive to heat; however it was stable for 6 months under accelerate conditions and 12 months under long-term test condition. Conclusion： The preparation of freeze-dried kit of Cys-Annexin V has advantages of easy manipulation with good stability,and its quality control is a reliable.

关 键 词：锝[99mTc]标记 膜联蛋白V变体 冻干 质量控制 

分 类 号：R943.3[医药卫生—药剂学]