激光粒度仪在评价吸附无细胞百白破疫苗颗粒一致性及稳定性中的应用  

Application of laser particle size analyzer to evaluation of batch consistency and stability of adsorbed DTaP vaccine

作　　者：李喆 黄小艳 吴燕 宋晓红 龙珍 李月琪 黄涛宏 马霄 LI Zhe;HUANG Xiao-yan;WU Yan;SONG Xiao-hong;LONG Zhen;LI Yue-qi;HUANG Tao-hong;MA Xiao(Division of Diphtheria,Tetanus and Pertussis Vaccine and Toxins,Department of Biological Product Control,National Institute for Food and Drug Control,Beijing 102629,China)

机构地区：[1]中国食品药品检定研究院生物制品检定所百白破疫苗与毒素室,北京102629 [2]岛津企业管理(中国)有限公司,北京100020

出　　处：《中国生物制品学杂志》2020年第7期818-823,共6页Chinese Journal of Biologicals

基　　金：国家自然科学基金青年科学基金(81701978);国家科技重大专项重大新药创制(2018ZX09738005)。

摘　　要：目的应用激光粒度仪评价吸附无细胞百白破疫苗(以下简称百白破疫苗)颗粒粒径及分布的一致性和稳定性。方法使用激光粒度仪SALD-7500nano,采用批式池和高浓度池系统测定百白破疫苗、抗原蛋白及铝佐剂颗粒粒径及分布;选取不同中值粒径(D50)聚苯乙烯微球验证仪器的准确性及重现性;采用该仪器分析铝佐剂(A及B厂家)对百白破抗原蛋白的吸附效果,评价百白破疫苗(A、B及C厂家)颗粒粒径一致性(批内及批间)及稳定性(长期、加速及机械刺激)。结果D50分别为0.05、1及25μm的聚苯乙烯微球有证标准物质重复测定6次D50的相对误差(relative error,RE)<0.05%,RSD<5%,激光粒度仪的准确性及重现性良好。A及B厂家疫苗抗原蛋白颗粒粒径分布在30~300 nm范围,铝佐剂颗粒粒径分布在1~30μm之间,疫苗颗粒粒径在1~300μm范围,表明铝佐剂对抗原蛋白吸附效果良好。4批A厂家疫苗批间颗粒D50的CV为37%,A及B厂家疫苗批内颗粒D50的CV均<5%,表明疫苗颗粒批间有一定差别,批内一致性良好。A及B厂家疫苗4℃保存0、30及60 d颗粒D50的CV均小于2%,表明疫苗长期储存稳定性良好;37℃保存1 d疫苗颗粒粒径变化较小,-20℃明显增大;百白破疫苗经批式池于连续搅拌120 min后静置12 h,A及C厂家疫苗颗粒D50的CV分别为1.2%和>10%,表明搅拌过程对疫苗颗粒粒径与分布存在一定影响。结论激光粒度仪系统的准确性及重现性良好,适用于百白破疫苗颗粒的一致性和稳定性评价。Objective To evaluate the batch consistency and stability of adsorbed DTaP vaccine by laser particle size analyzer.Methods The particle size(D50)and distribution of DTaP vaccine,antigen protein and aluminum adjuvant particles were measured by the batch cell and high concentration cell systems in SALD-7500 nano laser particle size analyzer.The analyzer was verified for accuracy and reproducibility with polystyrene microspheres at various median particles(D50),by which the adsorption effect of DTaP antigen protein by aluminum adjuvant from manufacturers A and B were analyzed,while the consistency of particle sizes of DTaP vaccine from manufacturers A,B and C in intra-and interassays and the long-term,accelerated and mechanical stimulation stabilities were evaluated.Results The relative errors of six test results of standard polystyrene microparticles at D50 of 0.05,1 and 25μm were less than 0.05%,while the RSD was less than 5%,indicating high accuracy and good reproducibility of the laser particle size analyzer.The particle sizes of antigenic protein from manufacturers A and B were 30~300 nm,while those of aluminum adjuvant were 1~30μm,and those of the vaccine were 1~300μm,indicating that antigen protein was well adsorbed by the aluminum adjuvant.The CV of D50 of four batches of DTaP vaccine from manufacturer A in inter-assay was 37%,while those from manufacturers A and B in intra-assay were less than 5%,indicating a certain difference between different batches and good consistency within the same batch.All the CVs of D50 of vaccine from manufacturers A and B after storage at 4℃for 0,30 and 60 d were less than 2%,indicating high stability after long-term storage.However,the D50 of vaccine after storage at 37℃for 1 d showed slight change while increased significantly at-20℃.After storage in batch cell in continuously stirring for 120 min and in stationary state for 12 h,the CVs of D50 of vaccines from manufacturers A and C were1.2%and more than 10%respectively,indicating a certain effect of stirring on size

关 键 词：激光粒度仪 吸附无细胞百白破疫苗 粒径 一致性 稳定性 

分 类 号：R917[医药卫生—药物分析学]