Article(id=1236369019297649484, tenantId=1146029695717560320, journalId=1235980733773295621, issueId=1236369012058288373, 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=1678291200000, acceptedDateStr=2023-03-09, onlineDate=1772702944825, onlineDateStr=2026-03-05, pubDate=1693324800000, pubDateStr=2023-08-30, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1772702944825, onlineIssueDateStr=2026-03-05, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1772702944825, creator=13701087609, updateTime=1772702944825, updator=13701087609, issue=Issue{id=1236369012058288373, tenantId=1146029695717560320, journalId=1235980733773295621, year='2023', volume='32', issue='16', pageStart='1593', pageEnd='1696', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1772702943100, creator=13701087609, updateTime=1772703272680, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1236370394492490265, tenantId=1146029695717560320, journalId=1235980733773295621, issueId=1236369012058288373, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1236370394496684570, tenantId=1146029695717560320, journalId=1235980733773295621, issueId=1236369012058288373, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=1622, endPage=1628, ext={EN=ArticleExt(id=1236369019582862192, articleId=1236369019297649484, tenantId=1146029695717560320, journalId=1235980733773295621, language=EN, title=Establishment and preliminary validation of a method for detecting the titer of anti-adeno-associated virus 8 neutralizing antibody in monkey plasma, columnId=null, journalTitle=Chinese Journal of New Drugs, columnName=null, runingTitle=null, highlight=null, articleAbstract=
Objective: To develop a highly sensitive, efficient, and stable cell-based platform for detection of anti-adeno-associated virus 8 (AAV8) neutralizing antibody titer in monkey plasma and perform preliminary validation of the relevant performance of the method.
Methods: In vitro, an equal volume of diluted monkey plasma was incubated with AAV8-Luciferase and then the mixture was added to a 96-well plate lined with HEK293 cells and incubated overnight to infect HEK293 cells; after cell lysis, the cells were added to a firefly luciferase reporter gene assay and the fluorescence signal (RLU) was detected using ELISA to assess the plasma levels of anti-AAV8 neutralising antibodies and subsequently obtain the corresponding antibody titres. The assay's sensitivity (LOD) and titer reproducibility, precision, method robustness and sample stability were preliminary were preliminary validated.
Results: A highly sensitive, efficient and stable method for the detection of anti-AAV8 neutralizing antibody titer in monkey plasma at the cellular level was developed and preliminarily validated. The LOD value of the method was 10.36 ng·mL-1. The repeatability, precision and method robustness of the titer assay were good, and the positive control samples were stable after being placed at room temperature and 2 ℃~8 ℃ for 24 h and after freeze-thawing for 6 times at -65 ℃~-90 ℃/room temperature.
Conclusion: It has been preliminarily validated that the method can be used for the analysis and detection of anti-AAV8 neutralizing antibody titer in monkey plasma, which is useful for grasping the process of drug treatment and adjusting the treatment protocol in time and is suggestive for the development of AAV8 serotype gene therapy drugs and individualized treatment.
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目的： 建立一种基于细胞平台的高灵敏度、高效、稳定的猴血浆中抗腺相关病毒8（AAV8）中和抗体滴度的检测方法，并对该方法进行相关性能的初步确认。
方法： 在体外将等体积稀释后的猴血浆与AAV8-Luciferase孵育后，取混合液加入铺有HEK293细胞的96孔板中，孵育过夜以感染HEK293细胞；细胞裂解后加入萤火虫荧光素酶报告基因检测试剂，使用酶标仪检测荧光信号（RLU），以评估血浆中抗AAV8中和抗体含量，继而得到相应的抗体滴度。该研究对检测方法的检测灵敏度（LOD）及滴度实验重复性、精密度、方法稳健性和样品稳定性等性能进行初步确认。
结果： 建立了高灵敏度、高效、稳定的细胞水平检测猴血浆中抗AAV8中和抗体滴度的方法并完成初步确认。该方法LOD值为10.36 ng·mL-1，滴度实验重复性、精密度及方法稳健性良好，阳性对照样品室温放置24 h、2 ℃~8 ℃放置24 h、经-65 ℃~-90 ℃/室温冻融6次均保持稳定。
结论： 经初步确认，该方法可以用于猴血浆中抗AAV8中和抗体滴度的分析检测，利于掌握药物治疗进程，及时调整治疗方案，对AAV8血清型基因治疗药物的研发及个体化治疗有一定提示作用。
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, authorsList=陶泽萍, 曹艳容, 许波华, 周璐, 赵小平)}, authors=[Author(id=1236369021029897207, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236369019297649484, orderNo=0, firstName=null, middleName=null, lastName=null, nameCn=null, orcid=null, stid=null, country=null, authorPic=null, dead=0, email=1490422375@qq.com, emailSecond=null, emailThird=null, correspondingAuthor=0, authorType=1, ext={EN=AuthorExt(id=1236369021394800658, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236369019297649484, authorId=1236369021029897207, language=EN, stringName=Ze-ping TAO, firstName=Ze-ping, middleName=null, lastName=TAO, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=
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1College of Chemistry and Chemical Engineering, Shanghai University of Engineering Science, Shanghai 201620, China
2Shanghai Innostar Biotech Co., Ltd., Shanghai 201203, China
3Nantong InnoStar Biotech Co., Ltd., Haimen 226133, China, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null), CN=AuthorExt(id=1236369021499658266, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236369019297649484, authorId=1236369021029897207, language=CN, stringName=陶泽萍, firstName=null, middleName=null, lastName=null, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=
1, 2, 3, address=
1上海工程技术大学化学化工学院，上海 201620
2上海益诺思生物技术股份有限公司，上海 201203
3益诺思生物技术南通有限公司，海门 226133, bio={"content":"
陶泽萍，女，硕士研究生，主要从事药理学及药动学研究。E-mail: 1490422375@qq.com。
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陶泽萍，女，硕士研究生，主要从事药理学及药动学研究。E-mail: 1490422375@qq.com。
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2上海益诺思生物技术股份有限公司，上海 201203)]), AuthorCompany(id=1236369020757267422, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236369019297649484, xref=3, ext=[AuthorCompanyExt(id=1236369020765656031, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236369019297649484, companyId=1236369020757267422, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=
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3益诺思生物技术南通有限公司，海门 226133)])]), Author(id=1236369021600321573, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236369019297649484, orderNo=1, 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=1236369021780676659, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236369019297649484, authorId=1236369021600321573, language=EN, stringName=Yan-rong CAO, firstName=Yan-rong, middleName=null, lastName=CAO, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=
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4Shanghai Vitalgen BioPharma Co., Ltd., Shanghai 201210, China, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null), CN=AuthorExt(id=1236369021898117180, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236369019297649484, authorId=1236369021600321573, language=CN, stringName=曹艳容, firstName=null, middleName=null, lastName=null, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=
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4上海天泽云泰生物医药有限公司，上海 201210, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null)}, companyList=[AuthorCompany(id=1236369020883096552, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236369019297649484, xref=4, ext=[AuthorCompanyExt(id=1236369020895679465, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236369019297649484, companyId=1236369020883096552, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=
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4上海天泽云泰生物医药有限公司，上海 201210)])]), Author(id=1236369022044917835, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236369019297649484, 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=1236369022191718489, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236369019297649484, authorId=1236369022044917835, language=EN, stringName=Bo-hua XU, firstName=Bo-hua, middleName=null, lastName=XU, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=
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3Nantong InnoStar Biotech Co., Ltd., Haimen 226133, China, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null), CN=AuthorExt(id=1236369022443376749, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236369019297649484, authorId=1236369022044917835, language=CN, stringName=许波华, firstName=null, middleName=null, lastName=null, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=
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3益诺思生物技术南通有限公司，海门 226133, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null)}, companyList=[AuthorCompany(id=1236369020757267422, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236369019297649484, xref=3, ext=[AuthorCompanyExt(id=1236369020765656031, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236369019297649484, companyId=1236369020757267422, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=
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3益诺思生物技术南通有限公司，海门 226133)])]), Author(id=1236369022602760315, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236369019297649484, 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=1236369022732783756, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236369019297649484, authorId=1236369022602760315, language=EN, stringName=Lu ZHOU, firstName=Lu, middleName=null, lastName=ZHOU, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=
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2Shanghai Innostar Biotech Co., Ltd., Shanghai 201203, China, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null), CN=AuthorExt(id=1236369022837641373, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236369019297649484, authorId=1236369022602760315, language=CN, stringName=周璐, firstName=null, middleName=null, lastName=null, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=
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2上海益诺思生物技术股份有限公司，上海 201203, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null)}, companyList=[AuthorCompany(id=1236369020610466770, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236369019297649484, xref=2, ext=[AuthorCompanyExt(id=1236369020614661076, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236369019297649484, companyId=1236369020610466770, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=
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2上海益诺思生物技术股份有限公司，上海 201203)])]), Author(id=1236369022938304680, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236369019297649484, orderNo=4, firstName=null, middleName=null, lastName=null, nameCn=null, orcid=null, stid=null, country=null, authorPic=null, dead=0, email=xp.zhao@vitalgen.com, emailSecond=null, emailThird=null, correspondingAuthor=0, authorType=1, ext={EN=AuthorExt(id=1236369023085105335, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236369019297649484, authorId=1236369022938304680, language=EN, stringName=Xiao-ping ZHAO, firstName=Xiao-ping, middleName=null, lastName=ZHAO, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=
1, 4, address=
1College of Chemistry and Chemical Engineering, Shanghai University of Engineering Science, Shanghai 201620, China
4Shanghai Vitalgen BioPharma Co., Ltd., Shanghai 201210, China, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null), CN=AuthorExt(id=1236369023202545861, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236369019297649484, authorId=1236369022938304680, language=CN, stringName=赵小平, firstName=null, middleName=null, lastName=null, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=
1, 4, address=
1上海工程技术大学化学化工学院，上海 201620
4上海天泽云泰生物医药有限公司，上海 201210, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null)}, companyList=[AuthorCompany(id=1236369020476249023, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236369019297649484, xref=1, ext=[AuthorCompanyExt(id=1236369020484637633, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236369019297649484, companyId=1236369020476249023, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=
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