抗体制剂乳光现象与分子粒径及聚集倾向性研究

Opalescence appearance of antibody formulation correlation with molecular size or aggregation propensity

目的:对抗体制剂乳光现象与分子粒径、纯度及聚集倾向性的相关性进行系统研究。方法:通过研究多种抗体的乳光现象,其中包含不同浓度、pH/缓冲盐及辅料对乳光现象的影响,以及温度、光照、反复冻融及振荡等环境因素的影响,分析乳光现象与分子粒径、纯度(聚集体)、扩散相互作用指数(K_(D))及聚集温度(T_(agg))之间的关联。结果:抗体制剂乳光与分子粒径大小呈正相关,乳光程度的加深表明分子粒径有所增加且分子聚集倾向性增加,如C分子在高温条件下,乳光现象明显加深,浊度由3.3 NTU变为13.6 NTU,粒径由13.1 nm变为40.6 nm,K_(D)由正变负,T_(agg)由59.6℃变为50℃以下。结论:缓冲体系、pH及辅料等处方因素,以及温度等条件均会影响抗体制剂的乳光特性,乳光程度增加,预示制剂稳定性降低,更容易发生聚集。本研究为抗体药物制剂生产中乳光的检测评价提供重要参考。

Objective:To explore the correlations between opalescence and molecular size,purity or aggregation tendency in the antibody formulation.Methods:Studied on the opalescence phenomena of various antibodies,in⁃cluding the influence of different concentrations,pH/buffer,excipients,as well as the influence of environmental factors such as temperature,light,freeze-thaw and agitation,and the correlations had been analyzed between o⁃palescence and molecular size,purity(aggregation),K_(D)or T_(agg).Results:The degree of antibody opalescence was positively correlated with the molecular size,the deepening of the opalescence indicated that the molecular size was increased and the more tendency of molecular aggregation.For example,the opalescence of C molecule obviously enhanced under high temperature,and the result of turbidity changed from 3.3 NTU to 13.6 NTU,the molecular size changed from 13.1 nm to 40.6 nm,and the K_(D)changed from positive to negative.The T_(agg)changed from 59.6℃to below 50℃.Conclusion:The opalescence can be affected by buffer system,pH,excipients and ambient temperature,the degree of opalescence increased,indicating the stability become worse and the molecules prone to aggregate.This study provides important reference for the analysis and evaluation of opalescence in the antibody formulation development.