Article(id=1236330552370982916, tenantId=1146029695717560320, journalId=1235980733773295621, issueId=1236330549866975405, articleNumber=null, orderNo=null, doi=null, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=null, receivedDateStr=null, revisedDate=null, revisedDateStr=null, acceptedDate=1667318400000, acceptedDateStr=2022-11-02, onlineDate=1772693773596, onlineDateStr=2026-03-05, pubDate=1697299200000, pubDateStr=2023-10-15, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1772693773596, onlineIssueDateStr=2026-03-05, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1772693773596, creator=13701087609, updateTime=1772693773596, updator=13701087609, issue=Issue{id=1236330549866975405, tenantId=1146029695717560320, journalId=1235980733773295621, year='2023', volume='32', issue='19', pageStart='1905', pageEnd='2008', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1772693772998, creator=13701087609, updateTime=1772695059296, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1236335945046348400, tenantId=1146029695717560320, journalId=1235980733773295621, issueId=1236330549866975405, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1236335945046348401, tenantId=1146029695717560320, journalId=1235980733773295621, issueId=1236330549866975405, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=1987, endPage=1992, ext={EN=ArticleExt(id=1236330552589086736, articleId=1236330552370982916, tenantId=1146029695717560320, journalId=1235980733773295621, language=EN, title=Preparation and quality analysis of intravenous human immunoglobulin by chromatography combined with low-temperature ethanol, columnId=null, journalTitle=Chinese Journal of New Drugs, columnName=null, runingTitle=null, highlight=null, articleAbstract=
Objective: The study aims to prepare human immunoglobulin by chromatography combined with low-temperature ethanol, detect the effect of impurities removal in the preparation process and the effect of chromatography on product quality.
Methods: Human immunoglobulin (pH 4) was prepared by one-step anion-exchange chromatography from FII precipitated from healthy human plasma. It was further separated and purified by low-temperature ethanol-protein separation. The distribution of molecular size, IgA content, IgG content and IgG subclasses distribution, protein secondary structure of IgG were determined and analyzed. The effect of IgA and IgM removal of intravenous human immunoglobulin (pH 4) prepared by low-temperature ethanol protein separation and anion exchange purification was compared.
Results: The precipitation of component II was dissolved by 5-fold volume of water for injection, and the dissolution rate of IgG was higher after stirring for 2 h. The content of IgA in intravenous human immunoglobulin (pH 4) prepared by anion exchange chromatography combined with low-temperature ethanol was controlled below 100 μg·mL-1, consistent with the distribution of molecular size and IgG isoforms of commercial products. The virus inactivation flux of nanofilms (20 nm) was increased.
Conclusion: Anion exchange chromatography can reduce the residual amount of IgA, IgM and other impurities in IVIG products, improve the permeability of nano-film (20 nm), and has no significant effect on the key quality of IVIG products.
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目的： 层析结合低温乙醇法制备静脉注射人免疫球蛋白（IVIG），检测制备过程中杂质去除效果及层析对制品质量的影响。
方法： 本实验系用健康人血浆，采用低温乙醇蛋白分离法分离纯化得到的组分II（以下简称FII）沉淀作为原料，经过一步阴离子交换层析制备IVIG（pH 4），检测IVIG制品分子量大小分布、免疫球蛋白（Ig）A含量、IgG含量及IgG亚型分布、蛋白质二级结构，采用孔径为20 nm的纳滤膜过滤，对比分析低温乙醇蛋白分离法及联合阴离子交换纯化工艺制备的IVIG（pH4）制品的IgA，IgM去除效果及纳米膜的通量。
结果： FII沉淀经5倍注射用水溶解，搅拌2 h后的IgG溶出率较高，阴离子交换层析结合低温乙醇法制备的IVIG（pH 4）制品中IgA含量控制在100 μg·mL-1以内，提高了纳米膜（20 nm）病毒灭活的通量，与市售产品IgG亚型分布、二级结构一致，分子大小分布符合《中华人民共和国药典》规定。
结论： 阴离子交换层析可以降低产品中IgA，IgM等杂质的残留量，提高了纳米膜（20 nm）的通透性，且对IVIG制品关键质量无显著影响。
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周雁翔，女，副研究员，主要从事血液制品研发。E-mail: 1403119938@qq.com。
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周雁翔，女，副研究员，主要从事血液制品研发。E-mail: 1403119938@qq.com。
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