摘要
目的采用离子交换层析技术从人血浆冷沉淀中分离纯化人纤维蛋白原(human fibrinogen,Fg)。方法采用离子交换层析技术从人血浆冷沉淀中连续制备3批次Fg制品,并根据《中华人民共和国药典》2020版(三部)(简称《中国药典》)的要求检测相关质量指标,首先对Fg制品进行了S/D病毒灭活前后蛋白含量、Fg含量和纯度指标差异性的测定,然后计算层析、超滤、除菌过滤和干热病毒灭活后各级的回收率,再进行干热病毒灭活前后蛋白含量、Fg含量、纯度和凝固活力指标差异性的检测,最后对成品进行物理检查和化学检定分析,并与国内同类产品进行比较。结果3批次实验结果显示,S/D处理对Fg无显著性影响(P>0.05);Fg各级回收率均值分别为69.0%、59.1%、54.4%和49.0%;干热病毒灭活处理对Fg无显著性影响(P>0.05);Fg物理检查(成品外观、复溶时间、可见异物和渗透压摩尔浓度)结果均合格;化学检定结果均在《中国药典》规定范围内,其中,水分含量为(1.6±0.1)%,TNBP和Tween-80残留量分别为(0.1±0.0)μg/mL和(8.0±2.6)μg/mL,成品的纯度为(83.4±1.3)%,盐酸精氨酸、枸橼酸离子和氯离子含量分别为(35.7±1.5)g/L、(28.0±1.0)mmol/L和(294.7±2.3)mmol/L,HBsAg检测结果均为阴性。Fg成品的关键质量指标与国内同类产品比较均能满足《中国药典》的要求。结论离子交换层析技术能从人血浆冷沉淀中分离纯化出安全、稳定和可靠的Fg制品。
Objective Study of the isolation and purification of human fibrinogen(Fg)from human plasma cryoprecipitate by ion exchange chromatography.Methods Three batches of Fg products were successively prepared from human plasma cryoprecipitate by ion exchange chromatography,and the relevant quality indexes were tested according to the requirements of Pharmacopoeia of the People's Republic of China 2020 Edition(PartⅢ).Firstly,the Fg products were investigated for the variability of the indexes of protein content,Fg content,and purity before and after the inactivation of the S/D virus,and then the recoveries of the products were calculated after chromatography,ultrafiltration,sterilization filtration and dry heat virus inactivation.And then the difference of protein content,Fg content,purity and coagulation activity index before and after dry heat virus inactivation was studied,and finally analyzed the final products by physical examination and chemical testing and compared them with similar products in China.Results The results of 3 batches of experiments showed S/D treatment had no significant effect on Fg(P>0.05).The average recoveries of Fg were 69.0%,59.1%,54.4%and 49.0%,respectively.Dry heat virus inactivation had no significant effect on Fg(P>0.05),and the results of the physical examination of Fg(appearance,redissolution time,visible foreign bodies and osmotic pressure molar concentration)of Fg were all qualified,the results of chemical examination were all within the range of Pharmacopoeia of the People's Republic of China,in which the moisture content was(1.6±0.1)%,the residues of TNBP and Tween-80 were(0.1±0.0)μg/mL and(8.0±2.6)μg/mL respectively,and the purity of the final product was(83.4±1.3)%,the contents of argine hydrochloride,citric acid ion and chloride ion were(35.7±1.5)g/L,(28.0±1.0)mmol/L and(294.7±2.3)mmol/L,the test results of HBsAg were all negative.The key quality indicators of Fg final products can meet the requirements of Pharmacopoeia of People's Republic of China as well as similar domestic products.Conclusion Ion exchange chromatography enables the isolation and purification of safe,stable and reliable Fg products from human plasma cryoprecipitate.
作者
刘环
张良
毛馨茸
张伟
郭亮
江砚芳
LIU Huan;ZHANG Liang;MAO Xinrong;ZHANG Wei;GUO Liang;JIANG Yanfang(Blood Preparation Department,Sinopharm Shanghai Plasma-derived Biotherapies Co.,Ltd..,Shanghai 200051,China;不详)
出处
《微生物学免疫学进展》
CAS
2023年第6期42-47,共6页
Progress In Microbiology and Immunology
关键词
人纤维蛋白原
血浆冷沉淀
层析
分离纯化
化学物残留
纯度
Human fibrinogen
Plasma cryoprecipitate
Chromatography
Separation and purification
Chemical residue
Purity
作者简介
刘环(1993-),男,硕士研究生,医学生物学工程师,主要从事血液制品制剂工作;共享第一作者:张良(1981-),男,本科,医学生物学高级工程师,主要从事血液制品研发和生产工作;通信作者:江砚芳,医学生物学高级工程师,E-mail:jiangyanfang@sinopharm.com。
