Article(id=1322906834780168205, tenantId=1146029695717560320, journalId=1235980733773295621, issueId=1322887806737825858, 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=1629734400000, acceptedDateStr=2021-08-24, onlineDate=1793335167832, onlineDateStr=2026-10-30, pubDate=1634227200000, pubDateStr=2021-10-15, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1793335167832, onlineIssueDateStr=2026-10-30, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1793335167832, creator=13701087609, updateTime=1793335167832, updator=13701087609, issue=Issue{id=1322887806737825858, tenantId=1146029695717560320, journalId=1235980733773295621, year='2021', volume='30', issue='19', pageStart='1729', pageEnd='1824', issueExtLink='null', onlineDate='null', pubDate='1634227200000', pubDateStr='2021-10-15', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1793330631192, creator='13701087609', updateTime=1793337389212, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1322916151994495727, tenantId=1146029695717560320, journalId=1235980733773295621, issueId=1322887806737825858, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1322916151994495728, tenantId=1146029695717560320, journalId=1235980733773295621, issueId=1322887806737825858, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=1787, endPage=1793, ext={EN=ArticleExt(id=1322906834943746062, articleId=1322906834780168205, tenantId=1146029695717560320, journalId=1235980733773295621, language=EN, title=Development and application of a sandwich ELISA method for detection of N protein in SARS-CoV-2 inactivated vaccine, columnId=null, journalTitle=Chinese Journal of New Drugs, columnName=null, runingTitle=null, highlight=null, articleAbstract=
Objective: To develop a sandwich ELISA method for detection of N protein in SARS-CoV-2 inactivated vaccine, and to further validate and primarily apply it for N protein qutification.
Methods: The indirect ELISA method was used to assess the affinity of different anti-N protein antibodies by coating the recombinant N protein. Then the sandwich ELISA system was set up by using antibodies rN003 and rN002 as the capture and detection antibody, respectively. Also, the method for fully lysing virus to release all the N protein was confirmed. Validation of linear range, accuracy, precision, intermediate precision, specificity and LLD were obtained, and N protein content in 6 batches of SARS-CoV-2 inactivated vaccine was determined using this method.
Results: N protein content has good linearty within the concentration range of 0.018~0.75 μg·mL-1 (R2>0.98), with the recovery of 80%~120% and CV <10%. N protein concentrations for 6 batches of samples accounted for 10%~20% of the total protein. All samples showed a good uniformity.
Conclusions: The sandwich ELISA method for detection of N protein in SARS-CoV-2 inactivated vaccine was successfully developed with good linearty, accuracy, precision, intermediate precision, selectivity, and sensitivity, which can satisfy the N protein detection of SARS-CoV-2 inactivated vaccine.
, authors=Yu-xiu ZHAO
1, Zhi-fang YING
2, Yan-li SONG
1, Ai-ling LI
1, Yan-cen GUO
1, Yuan DONG
1, Yan LI
1, Hui WANG
1, authorsList=Yu-xiu ZHAO, Zhi-fang YING, Yan-li SONG, Ai-ling LI, Yan-cen GUO, Yuan DONG, Yan LI, Hui WANG, authorCompany=null, correspAuthors=Hui WANG, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, fund=null), CN=ArticleExt(id=1322906835178627087, articleId=1322906834780168205, tenantId=1146029695717560320, journalId=1235980733773295621, language=CN, title=新型冠状病毒灭活疫苗N蛋白含量双抗体夹心ELISA检测方法的建立与应用, columnId=1236330552693944345, journalTitle=中国新药杂志, columnName=生物医药前沿, runingTitle=null, highlight=null, articleAbstract=
目的： 建立新型冠状病毒（SARS coronavirus 2，SARS-CoV-2）灭活疫苗N蛋白含量双抗体夹心ELISA检测方法，并对其进行验证及初步应用。
方法： 首先包被重组N蛋白，采用间接ELISA方法测定不同抗体对N蛋白的亲和能力。再选取高亲和力的抗体rN003作为包被抗体，rN002作为检测抗体，建立双抗体夹心ELISA检测方法并确定病毒裂解方法。测定该方法的线性范围、准确度、精确度、中间精密度、特异性及灵敏度，并用该方法测定6批SARS-CoV-2灭活疫苗中N蛋白的含量。
结果： N蛋白含量在0.018~0.75 μg·mL-1范围内该方法呈现良好的线性关系（R2>0.98），回收率在80%~120%之间，CV<10%。6批样品N蛋白含量均占总蛋白的10%~20%，样品均一性较好。
结论： 本研究成功建立了SARS-CoV-2灭活疫苗N蛋白含量双抗体夹心ELISA检测方法，该方法具有良好的线性、准确度、精确度、中间精密度、特异性及灵敏度，能满足SARS-CoV-2灭活疫苗N蛋白含量的检测。
, authors=赵玉秀
1, 英志芳
2, 宋彦丽
1, 李爱灵
1, 郭彦岑
1, 董圆
1, 李艳
1, 王辉
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赵玉秀，女，副研究员，主要从事新冠疫苗研发。联系电话：（010）60963531，E-mail：zhao10306@126.com。
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