tANCHOR cell-based ELISA approach as a surrogate for antigen-coated plates to monitor specific IgG directed to the SARS-CoV-2 receptor binding domain

IF 2.5 Q3 BIOCHEMICAL RESEARCH METHODS Biology Methods and Protocols Pub Date : 2024-01-19 DOI:10.1093/biomethods/bpae001
Hubert Bernauer, Josef Maier, Norbert Bannert, Daniel Ivanusic
{"title":"tANCHOR cell-based ELISA approach as a surrogate for antigen-coated plates to monitor specific IgG directed to the SARS-CoV-2 receptor binding domain","authors":"Hubert Bernauer, Josef Maier, Norbert Bannert, Daniel Ivanusic","doi":"10.1093/biomethods/bpae001","DOIUrl":null,"url":null,"abstract":"\n Enzyme-linked immunosorbent assay (ELISA) systems use plates coated with peptides or expressed and purified proteins to monitor immunoglobulins derived from patient serum. However, there is currently no easy, flexible, and fast adaptive ELISA-based system for testing antibodies directed against new SARS-CoV-2 variants. In this study, we utilized the tANCHOR protein display system that provides a cell surface decorated with the receptor binding domain (RBD) to monitor specific antibodies derived from SARS-CoV-2 convalescent and vaccinated individuals directed against it. To test sera from vaccinees or convalescent individuals, only the RBD coding sequence needs to be cloned in the tANCHOR vector system and transfected into HeLa cells. Time-consuming protein expression, isolation, and purification followed by coating assay plates is not necessary. With this technique, the immune evasion of new SARS-CoV-2 variants from current vaccination regimes can be examined quickly and reliably.","PeriodicalId":36528,"journal":{"name":"Biology Methods and Protocols","volume":null,"pages":null},"PeriodicalIF":2.5000,"publicationDate":"2024-01-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biology Methods and Protocols","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1093/biomethods/bpae001","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
引用次数: 0

Abstract

Enzyme-linked immunosorbent assay (ELISA) systems use plates coated with peptides or expressed and purified proteins to monitor immunoglobulins derived from patient serum. However, there is currently no easy, flexible, and fast adaptive ELISA-based system for testing antibodies directed against new SARS-CoV-2 variants. In this study, we utilized the tANCHOR protein display system that provides a cell surface decorated with the receptor binding domain (RBD) to monitor specific antibodies derived from SARS-CoV-2 convalescent and vaccinated individuals directed against it. To test sera from vaccinees or convalescent individuals, only the RBD coding sequence needs to be cloned in the tANCHOR vector system and transfected into HeLa cells. Time-consuming protein expression, isolation, and purification followed by coating assay plates is not necessary. With this technique, the immune evasion of new SARS-CoV-2 variants from current vaccination regimes can be examined quickly and reliably.
查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
用基于 tANCHOR 细胞的 ELISA 方法替代抗原涂层板,监测针对 SARS-CoV-2 受体结合域的特异性 IgG
酶联免疫吸附试验(ELISA)系统使用涂有肽或表达和纯化蛋白质的平板来监测从病人血清中提取的免疫球蛋白。然而,目前还没有一种简便、灵活、快速的自适应 ELISA 系统来检测针对 SARS-CoV-2 新变种的抗体。在这项研究中,我们利用了 tANCHOR 蛋白显示系统,该系统提供了一个由受体结合域(RBD)装饰的细胞表面,用于监测来自 SARS-CoV-2 康复者和疫苗接种者的针对 SARS-CoV-2 的特异性抗体。要检测疫苗接种者或康复者的血清,只需在 tANCHOR 载体系统中克隆 RBD 编码序列并转染 HeLa 细胞。无需进行耗时的蛋白质表达、分离和纯化,然后再涂布检测板。有了这项技术,就可以快速可靠地检测当前疫苗接种方案中新的 SARS-CoV-2 变体的免疫逃避能力。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 去求助
来源期刊
Biology Methods and Protocols
Biology Methods and Protocols Agricultural and Biological Sciences-Agricultural and Biological Sciences (all)
CiteScore
3.80
自引率
2.80%
发文量
28
审稿时长
19 weeks
期刊最新文献
Optimizing Western blotting immunodetection: Streamlining antibody cocktails for reduced protocol time and enhanced multiplexing applications. Live cell fluorescence microscopy-an end-to-end workflow for high-throughput image and data analysis. A reproducible method to study traumatic injury-induced zebrafish brain regeneration. Cluster analysis identifies long COVID subtypes in Belgian patients. Unpacking unstructured data: A pilot study on extracting insights from neuropathological reports of Parkinson's Disease patients using large language models.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1