重组人白介素1受体拮抗剂(rhIL-1Ra)重复给药导致大鼠和食蟹猴产生结合抗体  

Anti-rhIL-1Ra Binding Antibody Detection in Rats and Cynomolgus Monkeys Post Repeated rhIL-1Ra Injection

作　　者：杨敏敏[1,2] 陆海波 韩伟[1] 郜尽[1] 

机构地区：[1]上海交通大学药学院,上海200240 [2]交晨生物医药技术(上海)有限公司,上海201203

出　　处：《上海交通大学学报（农业科学版）》2014年第4期14-18,共5页Journal of Shanghai Jiaotong University(Agricultural Science)

基　　金：上海市科委重点科技攻关项目(11431921300)

摘　　要：限制大分子药物使用的关键因素之一为诱导抗体产生。新药安全性评价时往往不得不采用多种动物评价人源蛋白的安全性,因此,测定动物产生抗体水平是正确评价大分子药物的前提。为了检验重组人白介素1受体拮抗剂(rhIL-1Ra)在动物体内的安全性,采用间接酶联免疫吸附测定法(ELISA)测定rhIL-1Ra结合抗体,以纯化抗原作为包被载体,加入供试品后再加酶标抗体,用底物显色并在酶标仪上读取结果。结果显示,抗原包被浓度为10μg/mL、酶标二抗工作浓度为1∶5 000、阳性对照工作浓度为1∶10 000、待检血清样品稀释度从1∶100开始较为合适。SD大鼠肌肉注射给予rhIL-1Ra 4周试验中,剂量分别为0、5、15和30mg/kg。结果表明,剂量≥5mg/kg时给药2周后可诱导产生结合抗体。食蟹猴每天1次连续给药4周,剂量分别采用0、2.5、7.5和15.0mg/kg,当剂量≥2.5mg/kg时可诱导产生结合抗体。Anti-rhlL-1Ra binding antibodies in serum were tested by enzyme-linked immunosorbent assay （ELISA）. The operation of the assay included pure antigen coating, sample addition, enzyme labeled antibody addition and quantifying the absorbance of substrate in a microplate reader. The results showed that the key indexes of the developed method were as follows.the concentration of the coated antigen was 10μg/mL, the titer of enzyme-labeled antibody was 1：5 000, the work concentration of the positive control serum was 1： 10 000, and the dilution of test serum began from 1： 100. SD rats were administrated with rhIL-1Ra by im at doses of 0,5,15 and 30 mg/kg for four weeks. The results of animal experiment showed that doses ≥ 5 mg/kg after two weeks of rhIL-1Ra administration induced rhIL-1Ra binding antibodies. Meanwhile,the similar test on Cynomolgus monkeys at doses of 0,2. 5,7. 5 and 15.0 mg/kg indicated that doses ≥2. 5 mg/kg induced binding antibodies.

关 键 词：重组人白介素1受体拮抗剂 结合抗体 酶联免疫吸附测定法 

分 类 号：R322.7[医药卫生—人体解剖和组织胚胎学] Q255[医药卫生—基础医学]