PCSK9单抗生物学活性测定方法的建立

发布时间：2018-06-02 16:40

本文选题：单克隆抗体 + 前蛋白转化酶枯草溶菌素(PCSK)　；参考：《中国新药杂志》2015年15期

【摘要】：目的:建立前蛋白转化酶枯草溶菌素9(PCSK9)单抗的生物学活性检测方法。方法:利用系列稀释的PCSK9单抗以剂量依赖性方式阻断PCSK9与低密度脂蛋白受体(LDLR)结合,增加低密度脂蛋白(LDL)在Hep G2细胞内摄取率,通过BODIPY类荧光染料标记低密度脂蛋白(BODIPY-LDL)检测胞内荧光强度来反映PCSK9单抗的生物学活性,并用PCSK9单抗参比品计算供试品相对百分效价。结果:PCSK9、PCSK9单抗参比品及供试品在该方法中均存在量效关系,且符合四参数方程式:Y=(A-D)/[1+(X/C)B]+D。利用该法对4批PCSK9单抗供试品进行检测,每批供试品重复试验3次,PCSK9单抗的相对百分效价的平均值在94.25%~110.77%之间,变异系数均小于15%。结论:该方法灵敏度高、重复性好、结果准确,可作为PCSK9单抗生物学活性测定的常规检测方法。
[Abstract]:Objective: to establish a method for the determination of bioactivity of proprotein converting enzyme lysogenin 9 PCSK9 monoclonal antibody. Methods: a series of diluted PCSK9 monoclonal antibodies were used to block the binding of PCSK9 to LDLR in a dose-dependent manner, and to increase the uptake rate of LDL in Hep G2 cells. The bioactivity of PCSK9 McAb was measured by BODIPY fluorescent dye labeled low density lipoprotein (LDL). The relative titer of PCSK9 McAb was calculated by using PCSK9 McAb reference material. Results there was a dose-effect relationship between the reference sample and the test sample of the McAb of 10% PCSK9 and PCSK9, which was in accordance with the four-parameter equation: 1 X% Cnb / [1 X / Cnb] D. the results showed that there was a dose-effect relationship between the reference sample and the tested sample in this method. Four batches of PCSK9 McAbs were tested by this method. The average value of the relative percent titer of each sample was 94.25% and 110.77%, and the coefficient of variation was less than 15%. Conclusion: this method is sensitive, reproducible and accurate. It can be used as a routine method for the determination of biological activity of PCSK9 monoclonal antibody.
【作者单位】：中国食品药品检定研究院卫生部生物技术产品检定方法及其标准化重点实验室;
【基金】：国家“重大新药创制”科技重大专项(2014ZX09304311-001;2012ZX09304010)
【分类号】：R927.1

【共引文献】