Development of a SARS-CoV-2-specific biosensor for antigen detection using scFv-Fc fusion proteins

被引：117

作者：

Kim, Hye-Yeon ^{[1
,2
]}

Lee, Jong-Hwan ^{[1
]}

Kim, Mi Jeong ^{[1
,2
]}

Park, Sun Cheol ^{[1
,2
]}

Choi, Minsuk ^{[1
]}

Lee, Wonbin ^{[1
,2
]}

Ku, Keun Bon ^{[1
]}

Kim, Bum Tae ^{[1
]}

Park, Edmond Changkyun ^{[1
,2
,3
]}

Kim, Hong Gi ^{[1
]}

Kim, Seung Il ^{[1
,2
,3
]}

机构：

[1] Korea Res Inst Chem Technol, Ctr Convergent Res Emerging Virus Infect, Daejeon 34114, South Korea

[2] Korea Basic Sci Inst, Res Ctr Bioconvergence Anal, Cheongju 28119, South Korea

[3] Univ Sci & Technol, Dept Bioanal Sci, Daejeon 34113, South Korea

来源：

BIOSENSORS & BIOELECTRONICS | 2021年 / 175卷

基金：

新加坡国家研究基金会;

关键词：

COVID-19; SARS-CoV-2; Lateral flow immunoassay; Rapid diagnostic test; Antibody; ANTIBODIES;

D O I：

10.1016/j.bios.2020.112868

中图分类号：

Q6 [生物物理学];

学科分类号：

071011 ;

摘要：

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 x 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.

引用

页数：8