摘要
目的调查活化凝血因子Ⅺ(activated coagulation factorⅪ,FⅪa)在静注人免疫球蛋白各个生产环节中的分布及含量,分析其在生产过程中的主要形成过程、去除过程及其影响因素。方法采用底物显色法对合并血浆、组分Ⅰ+Ⅱ+Ⅲ(fractionⅠ+Ⅱ+Ⅲ,FⅠ+Ⅱ+Ⅲ)上清液、FⅠ+Ⅱ+Ⅲ沉淀溶解液、FⅠ+Ⅲ沉淀溶解液、FⅠ+Ⅲ上清液、FⅡ沉淀溶解液、FⅡ精制滤过液以及超滤配制后、低pH孵放后制品的FⅪa含量进行检测。添加硅藻土对合并血浆的FⅪ进行激活,将FⅠ+Ⅱ+Ⅲ沉淀溶解液采用双层滤纸过滤或离心进行FⅪa去除。结果FⅠ+Ⅱ+Ⅲ分离阶段合并血浆FⅪ被激活形成FⅪa,其激活量为合并血浆的2500倍。FⅠ+Ⅲ分离阶段可去除合并血浆FⅪ活化后形成的约99.8%的FⅪa。结论FⅠ+Ⅱ+Ⅲ分离阶段为FⅪa形成的主要阶段,该阶段硅藻土启动内源性凝血途径形成FⅪa;FⅠ+Ⅲ分离阶段为FⅪa去除的主要阶段,该阶段废弃吸附了FⅪa的硅藻土以去除工艺中形成的FⅪa;硅藻土为FⅪa形成与去除的主要影响因素。
Objective To investigate the distribution and content of activated coagulation factorⅪ(FⅪa)in various production processes of human immunoglobulin for intravenous injection,and analyze its main formation process,removal process and influencing factors in the production process.Methods Substrate chromogenic method was used to measure the content of FⅪa in the combined plasma,fractionⅠ+Ⅱ+Ⅲ(FⅠ+Ⅱ+Ⅲ)supernatant,FⅠ+Ⅱ+Ⅲprecipitate solution,FⅠ+Ⅲprecipitate solution,FⅠ+Ⅲsupernatant,FⅡprecipitate solution,FⅡrefined filtrate,product after preparation of ultrafiltration,and product after low pH incubation.Diatomite was added to activate the FⅪof the pooling plasma,and the FⅪa was removed from the FⅠ+Ⅱ+Ⅲprecipitation solution by double-layer filter paper filtration or centrifugation.Results The pooling plasma FⅪin the separation stage of FⅠ+Ⅱ+Ⅲwas activated to form FⅪa,and its activation amount was 2500 times of that from pooling plasma.The separation stage of FⅠ+Ⅲremoved about 99.8%of FⅪa formed after the activation of FⅪin the pooling plasma.Conclusions The separation stage of FⅠ+Ⅱ+Ⅲis the main stage of FⅪa formation,and at this stage,diatomite initiates the endogenous coagulation pathway to form FXIa.The separation stage of FⅠ+Ⅲis the main stage of FⅪa removal,and at this stage,the diatomite adsorbed with FXIa is discarded to remove the FXIa formed in the production process.Diatomite is the main influencing factor for the formation and removal of FXIa.
作者
周顺波
丁亚凌
田天丽
于海生
张姮婕
陈婕
陶迎秋
马晶
Zhou Shunbo;Ding Yaling;Tian Tianli;Yu Haisheng;Zhang Hengjie;Chen Jie;Tao Yingqiu;Ma Jing(Department of Quality Assurance,Chengdu Rongsheng Pharmaceuticals Co.,Ltd.,Chengdu 610219,China;NMPA Key Laboratory for Quality Control and Evaluation of Vaccines and Biological Products,Chengdu 611731,China)
出处
《国际生物制品学杂志》
CAS
2023年第6期333-337,共5页
International Journal of Biologicals
基金
国家药品监督管理局疫苗及生物制品质量控制与评价重点实验室开放课题(2022-KFKT-002)。
关键词
静注人免疫球蛋白
硅藻土
低温乙醇分离
低pH孵放
凝血因子Ⅺ
活化凝血因子Ⅺ
Human immunoglobulin for intravenous injection
Celite
Low temperature ethanol separation
Low pH incubation
Coagulation factorⅪ
Activated coagulation factorⅪ
