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Development of a SARS-CoV-2-specific biosensor for antigen detection using scFv-Fc fusion proteins
Coronavirus disease 2019 (COVID-19) is a newly emerged human infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In a global pandemic, development of a cheap, rapid, accurate, and easy-to-use diagnostic test is necessary if we are to mount an immediate response...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier B.V.
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7703470/ https://www.ncbi.nlm.nih.gov/pubmed/33281048 http://dx.doi.org/10.1016/j.bios.2020.112868 |
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author | Kim, Hye-Yeon Lee, Jong-Hwan Kim, Mi Jeong Park, Sun Cheol Choi, Minsuk Lee, Wonbin Ku, Keun Bon Kim, Bum Tae Changkyun Park, Edmond Kim, Hong Gi Kim, Seung Il |
author_facet | Kim, Hye-Yeon Lee, Jong-Hwan Kim, Mi Jeong Park, Sun Cheol Choi, Minsuk Lee, Wonbin Ku, Keun Bon Kim, Bum Tae Changkyun Park, Edmond Kim, Hong Gi Kim, Seung Il |
author_sort | Kim, Hye-Yeon |
collection | PubMed |
description | Coronavirus disease 2019 (COVID-19) is a newly emerged human infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In a global pandemic, development of a cheap, rapid, accurate, and easy-to-use diagnostic test is necessary if we are to mount an immediate response to this emerging threat. Here, we report the development of a specific lateral flow immunoassay (LFIA)-based biosensor for COVID-19. We used phage display technology to generate four SARS-CoV-2 nucleocapsid protein (NP)-specific single-chain variable fragment-crystallizable fragment (scFv-Fc) fusion antibodies. The scFv-Fc antibodies bind specifically and with high affinity to the SARS-CoV-2 NP antigen, but not to NPs of other coronaviruses. Using these scFv-Fc antibodies, we screened three diagnostic antibody pairs for use on a cellulose nanobead (CNB)-based LFIA platform. The detection limits of the best scFv-Fc antibody pair, 12H1 as the capture probe and 12H8 as the CNB-conjugated detection probe, were 2 ng antigen protein and 2.5 × 10(4) pfu cultured virus. This LFIA platform detected only SARS-CoV-2 NP, not NPs from MERS-CoV, SARS-CoV, or influenza H1N1. Thus, we have successfully developed a SARS-CoV-2 NP-specific rapid diagnostic test, which is expected to be a simple and rapid diagnostic test for COVID-19. |
format | Online Article Text |
id | pubmed-7703470 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Elsevier B.V. |
record_format | MEDLINE/PubMed |
spelling | pubmed-77034702020-12-01 Development of a SARS-CoV-2-specific biosensor for antigen detection using scFv-Fc fusion proteins Kim, Hye-Yeon Lee, Jong-Hwan Kim, Mi Jeong Park, Sun Cheol Choi, Minsuk Lee, Wonbin Ku, Keun Bon Kim, Bum Tae Changkyun Park, Edmond Kim, Hong Gi Kim, Seung Il Biosens Bioelectron Article Coronavirus disease 2019 (COVID-19) is a newly emerged human infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In a global pandemic, development of a cheap, rapid, accurate, and easy-to-use diagnostic test is necessary if we are to mount an immediate response to this emerging threat. Here, we report the development of a specific lateral flow immunoassay (LFIA)-based biosensor for COVID-19. We used phage display technology to generate four SARS-CoV-2 nucleocapsid protein (NP)-specific single-chain variable fragment-crystallizable fragment (scFv-Fc) fusion antibodies. The scFv-Fc antibodies bind specifically and with high affinity to the SARS-CoV-2 NP antigen, but not to NPs of other coronaviruses. Using these scFv-Fc antibodies, we screened three diagnostic antibody pairs for use on a cellulose nanobead (CNB)-based LFIA platform. The detection limits of the best scFv-Fc antibody pair, 12H1 as the capture probe and 12H8 as the CNB-conjugated detection probe, were 2 ng antigen protein and 2.5 × 10(4) pfu cultured virus. This LFIA platform detected only SARS-CoV-2 NP, not NPs from MERS-CoV, SARS-CoV, or influenza H1N1. Thus, we have successfully developed a SARS-CoV-2 NP-specific rapid diagnostic test, which is expected to be a simple and rapid diagnostic test for COVID-19. Elsevier B.V. 2021-03-01 2020-11-30 /pmc/articles/PMC7703470/ /pubmed/33281048 http://dx.doi.org/10.1016/j.bios.2020.112868 Text en © 2020 Elsevier B.V. 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 Kim, Hye-Yeon Lee, Jong-Hwan Kim, Mi Jeong Park, Sun Cheol Choi, Minsuk Lee, Wonbin Ku, Keun Bon Kim, Bum Tae Changkyun Park, Edmond Kim, Hong Gi Kim, Seung Il Development of a SARS-CoV-2-specific biosensor for antigen detection using scFv-Fc fusion proteins |
title | Development of a SARS-CoV-2-specific biosensor for antigen detection using scFv-Fc fusion proteins |
title_full | Development of a SARS-CoV-2-specific biosensor for antigen detection using scFv-Fc fusion proteins |
title_fullStr | Development of a SARS-CoV-2-specific biosensor for antigen detection using scFv-Fc fusion proteins |
title_full_unstemmed | Development of a SARS-CoV-2-specific biosensor for antigen detection using scFv-Fc fusion proteins |
title_short | Development of a SARS-CoV-2-specific biosensor for antigen detection using scFv-Fc fusion proteins |
title_sort | development of a sars-cov-2-specific biosensor for antigen detection using scfv-fc fusion proteins |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7703470/ https://www.ncbi.nlm.nih.gov/pubmed/33281048 http://dx.doi.org/10.1016/j.bios.2020.112868 |
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