Article(id=1236346589879521397, tenantId=1146029695717560320, journalId=1235980733773295621, issueId=1236346588398932061, 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=1660752000000, acceptedDateStr=2022-08-18, onlineDate=1772697597236, onlineDateStr=2026-03-05, pubDate=1685376000000, pubDateStr=2023-05-30, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1772697597236, onlineIssueDateStr=2026-03-05, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1772697597236, creator=13701087609, updateTime=1772697597236, updator=13701087609, issue=Issue{id=1236346588398932061, tenantId=1146029695717560320, journalId=1235980733773295621, year='2023', volume='32', issue='10', pageStart='969', pageEnd='1072', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1772697596882, creator=13701087609, updateTime=1772697715363, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1236347085411373506, tenantId=1146029695717560320, journalId=1235980733773295621, issueId=1236346588398932061, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1236347085411373507, tenantId=1146029695717560320, journalId=1235980733773295621, issueId=1236346588398932061, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=1000, endPage=1006, ext={EN=ArticleExt(id=1236346590085042308, articleId=1236346589879521397, tenantId=1146029695717560320, journalId=1235980733773295621, language=EN, title=Bioactivity determination of human insulin using in-cell Western, columnId=null, journalTitle=Chinese Journal of New Drugs, columnName=null, runingTitle=null, highlight=null, articleAbstract=
Objective: To study the bioactivity determination method of human insulin from transgenic cell using in-cell Western technology.
Methods: Using CHO-INSR B1284 cells as the target cell, different concentrations of human insulin were used as the stimulants. In-cell Western was used to measure the phosphorylation of human insulin receptor, and the bioactivity of human insulin was determined. According to the 2020 edition 9401 biological products bioactivity/potency determination method verification guiding principles in the General Rules of the fourth part of Chinese Pharmacopoeia, the established method was validated in terms of specificity, precision and durability. The national standard of recombinant human insulin was used as the benchmark to evaluate the relative potency of human insulin drug substance and other insulin analogues.
Results: Different concentrations of human insulin indicated effect on CHO-INSR B1284 cells in a dose-dependent manner. The samples of five potent concentrations were detected for 8 times. The geometric means of the relative potency were (63.76±4.38)%, (78.01±5.97)%, (99.52±4.62)%, (122.84±7.49)% and (151.71±8.81)%, respectively. The relative bias of five potency level were within ±5%, the corresponding geometric coefficient of variations (GCV,%) were lower than 11%, and the upper confidence limit of GCV were all lower than 20%. The bioactivity of human insulin and insulin analogues can be effectively detected using this method.
Conclusion: The in vitro bioactivity determination of human insulin using the In-cell Western was of good specificity, accuracy and precision, which can be used as a routine bioassay method of human insulin.
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目的： 利用in-cell Western（ICW）技术研究基于转基因细胞的人胰岛素生物学活性测定方法。
方法： 以CHO-INSR B1284为靶细胞，用不同浓度的人胰岛素刺激靶细胞，采用ICW技术检测人胰岛素受体酪氨酸磷酸化水平，进行人胰岛素的体外生物学活性测定；根据《中华人民共和国药典》2020年版通则9401“生物制品生物活性/效价测定方法”对该方法进行专属性、相对准确度、中间精密度、线性与范围等方法学验证；以重组人胰岛素国家标准品为参比品，采用本方法测定人胰岛素原料药及其他人胰岛素类似物的生物学活性相对效价。
结果： 不同浓度的人胰岛素作用于CHO-INSR B1284细胞后具有较好的剂量依赖性； 5个效价浓度的待测品经8次测定，相对效价的几何均值分别为：（63.76±4.38）%，（78.01±5.97）%，（99.52±4.62）%，（122.84±7.4）%，（151.71±8.81）%，相对偏倚均在±5%范围内，对应的几何变异系数（GCV，%）均<11%，GCV的置信上限均<20%；采用该方法能有效检测人胰岛素及其胰岛素类似物的效价。
结论： 利用ICW技术的人胰岛素体外生物学活性测定法专属性强、准确度高、精密度好，可作为人胰岛素生物学活性的常规检测方法。
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, authorsList=王绿音, 杨艳枫, 梁誉龄, 张慧, 李晶, 梁成罡)}, authors=[Author(id=1236346590730965162, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, orderNo=0, firstName=null, middleName=null, lastName=null, nameCn=null, orcid=null, stid=null, country=null, authorPic=null, dead=0, email=wanglvyin@nifdc.org.cn, emailSecond=null, emailThird=null, correspondingAuthor=0, authorType=1, ext={EN=AuthorExt(id=1236346590819045552, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, authorId=1236346590730965162, language=EN, stringName=Lv-yin WANG, firstName=Lv-yin, middleName=null, lastName=WANG, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=
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1Division of Hormone, National Institutes for Food and Drug Control, NHC Key Laboratory of Research on Quality and Standardization of Biotech Products, Beijing 102629, China, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null), CN=AuthorExt(id=1236346590911320246, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, authorId=1236346590730965162, language=CN, stringName=王绿音, firstName=null, middleName=null, lastName=null, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=
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王绿音，女，助理研究员，主要从事激素类药物质量控制研究。E-mail: wanglvyin@nifdc.org.cn。
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王绿音，女，助理研究员，主要从事激素类药物质量控制研究。E-mail: wanglvyin@nifdc.org.cn。
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1Division of Hormone, National Institutes for Food and Drug Control, NHC Key Laboratory of Research on Quality and Standardization of Biotech Products, Beijing 102629, China
2China Pharmaceutical University, Nanjing 211198, China, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null), CN=AuthorExt(id=1236346591154589898, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, authorId=1236346590995206331, language=CN, stringName=杨艳枫, firstName=null, middleName=null, lastName=null, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=
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1中国食品药品检定研究院激素室，国家卫生健康委员会生物技术产品检定方法及其标准化重点实验室，北京 102629)]), AuthorCompany(id=1236346590638690464, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, xref=2, ext=[AuthorCompanyExt(id=1236346590647079074, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, companyId=1236346590638690464, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=
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2中国药科大学，南京 211198)])]), Author(id=1236346591234281681, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, orderNo=2, firstName=null, middleName=null, lastName=null, nameCn=null, orcid=null, stid=null, country=null, authorPic=null, dead=0, email=null, emailSecond=null, emailThird=null, correspondingAuthor=0, authorType=1, ext={EN=AuthorExt(id=1236346591351722200, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, authorId=1236346591234281681, language=EN, stringName=Yu-ling LIANG, firstName=Yu-ling, middleName=null, lastName=LIANG, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=
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1Division of Hormone, National Institutes for Food and Drug Control, NHC Key Laboratory of Research on Quality and Standardization of Biotech Products, Beijing 102629, China, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null), CN=AuthorExt(id=1236346591427219681, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, authorId=1236346591234281681, language=CN, stringName=梁誉龄, firstName=null, middleName=null, lastName=null, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=
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1中国食品药品检定研究院激素室，国家卫生健康委员会生物技术产品检定方法及其标准化重点实验室，北京 102629, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null)}, companyList=[AuthorCompany(id=1236346590542221465, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, xref=1, ext=[AuthorCompanyExt(id=1236346590550610074, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, companyId=1236346590542221465, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=
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1中国食品药品检定研究院激素室，国家卫生健康委员会生物技术产品检定方法及其标准化重点实验室，北京 102629)])]), Author(id=1236346591536271589, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, orderNo=3, firstName=null, middleName=null, lastName=null, nameCn=null, orcid=null, stid=null, country=null, authorPic=null, dead=0, email=null, emailSecond=null, emailThird=null, correspondingAuthor=0, authorType=1, ext={EN=AuthorExt(id=1236346591628546289, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, authorId=1236346591536271589, language=EN, stringName=Hui ZHANG, firstName=Hui, middleName=null, lastName=ZHANG, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=
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1中国食品药品检定研究院激素室，国家卫生健康委员会生物技术产品检定方法及其标准化重点实验室，北京 102629, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null)}, companyList=[AuthorCompany(id=1236346590542221465, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, xref=1, ext=[AuthorCompanyExt(id=1236346590550610074, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, companyId=1236346590542221465, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=
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1中国食品药品检定研究院激素室，国家卫生健康委员会生物技术产品检定方法及其标准化重点实验室，北京 102629)])]), Author(id=1236346591813095679, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, orderNo=4, firstName=null, middleName=null, lastName=null, nameCn=null, orcid=null, stid=null, country=null, authorPic=null, dead=0, email=null, emailSecond=null, emailThird=null, correspondingAuthor=0, authorType=1, ext={EN=AuthorExt(id=1236346591917953287, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, authorId=1236346591813095679, language=EN, stringName=Jing LI, firstName=Jing, middleName=null, lastName=LI, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=
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1中国食品药品检定研究院激素室，国家卫生健康委员会生物技术产品检定方法及其标准化重点实验室，北京 102629, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null)}, companyList=[AuthorCompany(id=1236346590542221465, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, xref=1, ext=[AuthorCompanyExt(id=1236346590550610074, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, companyId=1236346590542221465, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=
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1中国食品药品检定研究院激素室，国家卫生健康委员会生物技术产品检定方法及其标准化重点实验室，北京 102629)])]), Author(id=1236346592094114071, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, orderNo=5, firstName=null, middleName=null, lastName=null, nameCn=null, orcid=null, stid=null, country=null, authorPic=null, dead=0, email=liangchenggang@nifdc.org.cn, emailSecond=null, emailThird=null, correspondingAuthor=0, authorType=1, ext={EN=AuthorExt(id=1236346592194777378, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, authorId=1236346592094114071, language=EN, stringName=Cheng-gang LIANG, firstName=Cheng-gang, middleName=null, lastName=LIANG, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=
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1Division of Hormone, National Institutes for Food and Drug Control, NHC Key Laboratory of Research on Quality and Standardization of Biotech Products, Beijing 102629, China, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null), CN=AuthorExt(id=1236346592266080548, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, authorId=1236346592094114071, language=CN, stringName=梁成罡, firstName=null, middleName=null, lastName=null, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=
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1中国食品药品检定研究院激素室，国家卫生健康委员会生物技术产品检定方法及其标准化重点实验室，北京 102629)])])], keywords=[Keyword(id=1236346592375132458, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, language=EN, orderNo=1, keyword=biological activity), Keyword(id=1236346592446435633, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, language=EN, orderNo=2, keyword=human insulin), Keyword(id=1236346592517738808, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, language=EN, orderNo=3, keyword=transgenic cells), Keyword(id=1236346592622596413, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, language=EN, orderNo=4, keyword=in-cell Western), Keyword(id=1236346592731648322, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, language=EN, orderNo=5, keyword=method validation), Keyword(id=1236346592802951494, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, language=CN, orderNo=1, keyword=生物学活性), Keyword(id=1236346592882643275, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, language=CN, orderNo=2, keyword=人胰岛素), Keyword(id=1236346592966529360, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, language=CN, orderNo=3, keyword=转基因细胞), Keyword(id=1236346593071386964, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, language=CN, orderNo=4, keyword=in-cell Western), Keyword(id=1236346593159467355, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, language=CN, orderNo=5, keyword=方法学验证)], refs=null, funds=[Fund(id=1236346593285296482, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, awardId=2018ZX09101001-003-004, language=CN, fundingSource=国家“重大新药创制”科技重大专项资助项目：创新生物技术药评价及标准化关键技术研究(2018ZX09101001-003-004), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1236346590542221465, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, xref=1, ext=[AuthorCompanyExt(id=1236346590550610074, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236346589879521397, companyId=1236346590542221465, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=
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