反复冻融对23价肺炎球菌荚膜多糖稳定性的影响  

Impact of multiple freeze-thaw cycles on stability of 23-valent pneumococcal capsular polysaccharide

作　　者：吕雪艳[1] 张磊[1] 陈思[1] 杨锣[1] 周学益 张立平 文崇艳 陈文敏[1] 陈亦鸣 张锐[1] LVXue-yan;ZHANG Lei;CHEN Si;YANG Luo;ZHOU Xue-yi;ZHANG Li-ping;WEN Chong-yan;CHEN Wen-min;CHEN Yi-ming;ZHANG Rui(Chengdu Institute of Biological Products Co.,Ltd.,Chengdu 610063,China)

机构地区：[1]成都生物制品研究所有限责任公司,成都610063

出　　处：《中国新药杂志》2020年第21期2515-2520,共6页Chinese Journal of New Drugs

摘　　要：目的:考察反复冻融对23种血清型(1,2,3,4,5,6B,7F,8,9N,9V,10A,11A,12F,14,15B,17F,18C,19A,19F,20,22F,23F和33F)肺炎球菌荚膜多糖的影响。方法:各型多糖由-20℃至室温(约25℃)反复冻融1~3次,每个时间点抽取样品进行多糖抗原性(速率比浊法)、分子大小(HPSEC-MALLS/RI分析法)和特异性基团含量(糖醛酸、甲基戊糖、氨基己糖和O-乙酰基含量等4项指标)的检测,将检测结果与正常使用水平数据(冻融1次)进行比较分析。结果:反复冻融1~3次,23种血清型肺炎球菌荚膜多糖的抗原性、分子量分布、特异性基团含量均未发生显著变化(P>0.05或含量与初始值相差<5%)。结论:反复冻融3次以内不会导致荚膜多糖的降解,各型多糖均表现出较高的稳定性。Objective: To study the impact of multiple freeze-thaw cycles on the stability of 23-valent( 1,2,3,4,5,6 B,7 F,8,9 N,9 V,10 A,11 A,12 F,14,15 B,17 F,18 C,19 A,19 F,20,22 F,23 F and 33 F)pneumococcal capsular polysaccharide. Methods: Freeze-thaw for 1 ~ 3 cycles were conducted on all types of polysaccharides at-20 ℃ to room temperature( approx. 25 ℃) . Samples were taken at each time point and polysaccharide antigenicity( rate nephelometry,RN),molecular size( HPSEC-MALLS/RI analysis) and the content of specific groups( uronic acid,methylpentose,aminohexose and O-acetyl) were tested. The results were compared to initial value( 1 freeze-thaw cycle) and analyzed. Results: after 1 ~ 3 cycles of freeze-thaw,there was no significant change( P > 0. 05 or 5% difference of content comparing to initial value) observed in antigenicity,molecular size distribution and content of specific groups of 23-valent pneumococcal capsular polysaccharide.Conclusion: Less than 3 cycles of freeze-thaw will not cause capsular polysaccharide degradation,and all types of polysaccharides show high stability within 3 cycles of freeze-thaw.

关 键 词：肺炎球菌荚膜多糖 反复冻融 速率比浊法 高效凝胶排阻色谱-多角度激光光散射仪/示差折光检测器联用技术 

分 类 号：R95[医药卫生—药学]