二硫键稳定的抗CD3/抗Pgp微型双功能抗体稳定性研究  

Stability analyze of the disulphide bond stabilized anti-CD3/anti-Pgp diabody

作　　者：杨铭[1] 程昕[1] 杜志荣[2] 王彦[3] 范冬梅[1] 刘娟妮[1] 王金宏[1] 纪庆[1] 高瀛岱[1] 

机构地区：[1]中国医学科学院&北京协和医学院血液学研究所,天津300020 [2]中国医学科学院&北京协和医学院基础医学研究所,北京100730 [3]中国医学科学院&北京协和医学院放射医学研究所,天津300192

出　　处：《中国免疫学杂志》2010年第12期1078-1081,共4页Chinese Journal of Immunology

基　　金：天津市科技计划资助项目(No.08ZCKFSH04100)

摘　　要：目的:研究二硫键稳定的抗CD3/抗Pgp微型双功能抗体(Diabody)在体外和体内的稳定性。方法:将抗CD3/抗Pgp Diabody置于37℃含0.2%人血清白蛋白(human serumalbumin,HSB)的PBS中,孵育不同时间后,用流式细胞术(FACS)检测其结合活性。建立K562/A02裸鼠移植瘤模型,用Cy5荧光标记试剂盒标记抗CD3/抗Pgp Diabody,通过尾静脉注射给荷瘤裸鼠,在小动物活体成像系统上动态观察荧光信号。结果:二硫键稳定的抗CD3/抗Pgp Diabody(Ds-diabdoy)体外孵育72小时后活性无明显下降,而改造前Diabody孵育1小时后活性即开始下降,24小时后活性完全丧失。Ds-diabdoy注射后72小时仍能在肿瘤部位检测到荧光信号,而改造前Diabody在注射后24小时肿瘤部位荧光信号消失。结论:Ds-diabdoy较改造前Diabody稳定性大大提高。Objective：To analyze the in vitro and in vivo stability of a disulphide anti-CD3/anti-Pgp diabody.Methods：The anti-CD3/anti-Pgp diabody was incubated in PBS which contained 0.2% HSB at 37℃ for different times,and then analyzed the binding activity by FACS.The diabody was labeled with the fluorescent dye Cy5.Labeled diabdoy was injected via tail veil into BALB/c nude mice bearing MDR（Multi-drug resistance）human xenografts.The fluorescence emission was recorded with a whole-body small-animal imaging system.Results：The remained binding activity of ds-diadody showed no obvious decrease after incubation for 72 h in vitro,while the remained binding activity of the parent diabody began to decrease after incubation for 1 h,and couldn＇t be detected after incubation for 24 h.Intense fluorescence could be detected for the ds-diabody at 72 h post-injection,while fluorescence for the parent diabody disappeared at 24 h post-injection.Conclusion：The data demonstrate that ds-diabody has improved stability compared with the parent diabody.

关 键 词：药物稳定性 DIABODY 二硫键 多药耐药 

分 类 号：R392-33[医药卫生—免疫学]