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Isolation and identification of an scFv antibody against nucleocapsid protein of SARS-CoV

To develop reagents for early diagnosis and therapeutic drugs against SARS-associated coronavirus (SARS-CoV), a large (3 × 10(9)) immunized human antibody library was constructed from peripheral blood mononuclear cells from six SARS convalescent patients. A single chain variable fragment antibody (N...

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Autores principales: Zhao, Aizhi, Qin, Weijun, Han, Yueheng, Wen, Weihong, Zhang, Wenhong, Lian, Zhonghui, Chen, Gang, Zhang, Zhuoli, Peng, Jianqiang, Wang, He, Guo, Yinglu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier Masson SAS. 2007
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7110486/
https://www.ncbi.nlm.nih.gov/pubmed/17548223
http://dx.doi.org/10.1016/j.micinf.2007.04.008
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author Zhao, Aizhi
Qin, Weijun
Han, Yueheng
Wen, Weihong
Zhang, Wenhong
Lian, Zhonghui
Chen, Gang
Zhang, Zhuoli
Peng, Jianqiang
Wang, He
Guo, Yinglu
author_facet Zhao, Aizhi
Qin, Weijun
Han, Yueheng
Wen, Weihong
Zhang, Wenhong
Lian, Zhonghui
Chen, Gang
Zhang, Zhuoli
Peng, Jianqiang
Wang, He
Guo, Yinglu
author_sort Zhao, Aizhi
collection PubMed
description To develop reagents for early diagnosis and therapeutic drugs against SARS-associated coronavirus (SARS-CoV), a large (3 × 10(9)) immunized human antibody library was constructed from peripheral blood mononuclear cells from six SARS convalescent patients. A single chain variable fragment antibody (N18) with high affinity against N protein of SARS-CoV was isolated. Sequence analysis revealed that the VL gene was composed of VL3 h (V lambda subgroup) and JL2 regions and the VH gene was composed of VH1-69 (VH1 subgroup), D2-15, D3-22 and JH6 regions. Soluble N18 antibody was expressed in Escherichia coli HB2151, purified by Ni–NTA affinity chromatography and verified by SDS-PAGE and Western blot. The potential application for early diagnosis was evaluated using N protein capture ELISA in which N18 antibody demonstrated high sensitive activity in detecting N protein of SARS-CoV. Finally, the potential usefulness of the N18 antibody in prophylaxis, vaccine design and therapy of SARS is discussed.
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spelling pubmed-71104862020-04-02 Isolation and identification of an scFv antibody against nucleocapsid protein of SARS-CoV Zhao, Aizhi Qin, Weijun Han, Yueheng Wen, Weihong Zhang, Wenhong Lian, Zhonghui Chen, Gang Zhang, Zhuoli Peng, Jianqiang Wang, He Guo, Yinglu Microbes Infect Article To develop reagents for early diagnosis and therapeutic drugs against SARS-associated coronavirus (SARS-CoV), a large (3 × 10(9)) immunized human antibody library was constructed from peripheral blood mononuclear cells from six SARS convalescent patients. A single chain variable fragment antibody (N18) with high affinity against N protein of SARS-CoV was isolated. Sequence analysis revealed that the VL gene was composed of VL3 h (V lambda subgroup) and JL2 regions and the VH gene was composed of VH1-69 (VH1 subgroup), D2-15, D3-22 and JH6 regions. Soluble N18 antibody was expressed in Escherichia coli HB2151, purified by Ni–NTA affinity chromatography and verified by SDS-PAGE and Western blot. The potential application for early diagnosis was evaluated using N protein capture ELISA in which N18 antibody demonstrated high sensitive activity in detecting N protein of SARS-CoV. Finally, the potential usefulness of the N18 antibody in prophylaxis, vaccine design and therapy of SARS is discussed. Elsevier Masson SAS. 2007-07 2007-04-21 /pmc/articles/PMC7110486/ /pubmed/17548223 http://dx.doi.org/10.1016/j.micinf.2007.04.008 Text en Copyright © 2007 Elsevier Masson SAS. All rights reserved. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.
spellingShingle Article
Zhao, Aizhi
Qin, Weijun
Han, Yueheng
Wen, Weihong
Zhang, Wenhong
Lian, Zhonghui
Chen, Gang
Zhang, Zhuoli
Peng, Jianqiang
Wang, He
Guo, Yinglu
Isolation and identification of an scFv antibody against nucleocapsid protein of SARS-CoV
title Isolation and identification of an scFv antibody against nucleocapsid protein of SARS-CoV
title_full Isolation and identification of an scFv antibody against nucleocapsid protein of SARS-CoV
title_fullStr Isolation and identification of an scFv antibody against nucleocapsid protein of SARS-CoV
title_full_unstemmed Isolation and identification of an scFv antibody against nucleocapsid protein of SARS-CoV
title_short Isolation and identification of an scFv antibody against nucleocapsid protein of SARS-CoV
title_sort isolation and identification of an scfv antibody against nucleocapsid protein of sars-cov
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7110486/
https://www.ncbi.nlm.nih.gov/pubmed/17548223
http://dx.doi.org/10.1016/j.micinf.2007.04.008
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