https://scholars.lib.ntu.edu.tw/handle/123456789/606014
標題: | Generation and characterization of a spike glycoprotein domain a-specific neutralizing single-chain variable fragment against porcine epidemic diarrhea virus | 作者: | Chang C.-Y Wang Y.-S Wu J.-F Yang T.-J Chang Y.-C Chae C Chang H.-W HUI-WEN CHANG YEN-CHEN CHANG |
關鍵字: | Neutralizing antibody;Porcine epidemic diarrhea virus;Single-chain variable fragment (scFv);agar;brilliant blue g;buffer;expi293;expifectamine;fluorescein isothiocyanate;horseradish peroxidase;imidazole;immunoglobulin heavy chain;immunoglobulin kappa chain;immunoglobulin lambda chain;kpl;peroxidase;phosphate buffered saline;r250;reagent;RNA directed DNA polymerase;single chain fragment variable antibody;smartscribe;sodium azide;sodium chloride;tissue plasminogen activator;virus spike protein;animal cell;antigen recognition;Article;controlled study;DNA sequence;epitope mapping;fast protein liquid chromatography;gene expression system;genetic transfection;hybridoma cell line;immobilized metal affinity chromatography;immune response;immunocytochemistry;immunofluorescence assay;immunoprecipitation;live cell imaging;nonhuman;polyacrylamide gel electrophoresis;protein analysis;protein domain;protein expression;reverse transcription polymerase chain reaction;RNA extraction;size exclusion chromatography;spike glycoprotein domain a;Western blotting | 公開日期: | 2021 | 卷: | 9 | 期: | 8 | 來源出版物: | Vaccines | 摘要: | The emergence of the genotype (G) 2 and re-emergence of the G1 porcine epidemic diarrhea virus (PEDV) has caused severe economic impacts in the past decade. Developments of efficient vaccines against new variants of PEDV have been challenging, not least because of the difficulties in eliciting mucosal and lactogenic immunity. A single-chain fragment variable (scFv) capable of efficient antigen recognition is an alternative to vaccination and treatment of a viral infection. In the present study, the variable regions of the light chain and the heavy chain of a G2b PEDV spike domain A (S1A)-specific neutralizing monoclonal antibody (mAb) were sequenced, constructed with a (G4S) x3 linker, and produced by a mammalian protein expression system. Our results demonstrated that the PEDV S1A domain scFv was able to bind to S proteins of both G1 and G2b PEDVs. Nevertheless, the scFv was only capable of neutralizing the homologous G2b PEDV but not the G1 PEDV. The binding ability of the G2b-specific neutralizing scFv was not able to predict the neutralizing ability toward heterologous PEDV. The anti-PEDV S1A scFv presented herein serves as a potential therapeutic candidate against the virulent G2b PEDV. ? 2021 by the authors. |
URI: | https://www.scopus.com/inward/record.uri?eid=2-s2.0-85112418286&doi=10.3390%2fvaccines9080833&partnerID=40&md5=a61d3c2087467f700d5d4130d9799667 https://scholars.lib.ntu.edu.tw/handle/123456789/606014 |
ISSN: | 2076393X | DOI: | 10.3390/vaccines9080833 |
顯示於: | 分子暨比較病理生物學研究所 |
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