抗白介素-13单克隆抗体生物学活性测定方法的建立

Establishment of A Method for the Biological Potency Detection of AntiInterleukin-13 Monoclonal Antibody

目的:建立抗白介素-13单克隆抗体(抗IL-13单抗)的生物学活性检测方法。方法:利用L-BEAS-2B细胞系,通过荧光素酶检测系统进行抗IL-13单抗的生物学活性检测,通过抗IL-13单抗参比品与样品对比以平行线分析的方法计算样品相对百分效价,并对该方法的精密性和准确性进行验证。结果:抗IL-13单抗在该方法中存在量效关系,在半对数坐标纸上呈平行的直线分布。6批抗IL-13单抗样品经3次测定,相对百分效价平均值在(91.10±1.10)%~(105.27±7.17)%之间,RSD均小于12%;1批样品经9次测定平均相对百分效价为(110.77±7.01)%,板间和日间RSD均小于7%;2批回收率样品经3次测定,回收率分别为(96.17±8.50)%和(91.53±15.46)%。结论:研究建立的抗IL-13单抗生物学活性检测方法准确性高,精密度及重复性好,可作为抗IL-13单抗生物学活性的常规检测方法。

Objective:To establish a method for the biological potency detection of anti-IL-13 monoclonal antibody.Methods:The biological potency of anti-IL-13 mAb was determined by Bright-Glo?Luciferase Assay System with L-BEAS-2 B cell lines acting as target cells and the relative percentage potency was calculated by comparing the samples with the reference with the method of parallel analysis.The developed method was validated in precision and accuracy.Results:Either the drug product or the reference of anti-IL-13 mAb showed a dose-response relationship by semi-log mode,presenting parallel straight lines.6 batches of drug products of the anti-IL-13 mAb had been tested for 3 times,the results showed that the mean of relative percentage potency was between(91.10±1.10)%and(105.27±7.17)%,with relative standard deviation less than 12%.The mean of relative percentage potency of one batch after 9 times of detections was(10.77±7.01)%,and neither the standard deviation of day to day nor that of plate to plate was higher 7%.The recovery samples of 2 batches had been detected for 3 times,the recoveries of which were(96.17±8.50)%and(91.53±15.46)%respectively.Conclusion:A method for detecting potency of anti-IL-13 mAb was successfully developed,which showed good reproducibility and accuracy and could be used as a routine potency detection for anti-IL-13 mAb.