Pig Sera-derived Anti-SARS-CoV-2 Antibodies in Surface Plasmon Resonance Biosensors
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Original Article
Pig Sera-derived Anti-SARS-CoV-2 Antibodies in Surface Plasmon Resonance Biosensors Ji-Hong Bong1, Tae-Hun Kim1, Jaeyong Jung1, Soo Jeong Lee1, Jeong Soo Sung1, Chang Kyu Lee1, Min-Jung Kang2, Hyun Ok Kim3 & Jae-Chul Pyun 1,* Received: 29 August, 2020 / Accepted: 18 September, 2020 / Published online: 22 October, 2020 ⒸThe Korean BioChip Society and Springer 2020
Abstract Anti-coronavirus disease-2019 (COVID-19; anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)) antibodies against nucleoprotein (NP) were purified from pig sera. Following the separation of the antibody fraction using a protein-A column, the final yield of the purified antibodies against SARSCoV-2 NPs was estimated to be 0.26 ± 0.05 % (absolute amount of 143.4 ± 25.2 ng, n=5) from 1 mL of pig sera. The binding activities of the isolated antibodies were confirmed using immunoassay and immunostaining. Based on the specific binding activity to NPs, a quantitative assay was performed using a surface plasmon resonance (SPR) biosensor. From the doseresponse curve, the binding constant (Kd) was calculated to be 185 pM and the limit of detection was estimated to be 1.02 pM. The SPR biosensor with the isolated antibodies against SARS-CoV-2 NPs was applied for the detection of SARS-CoV-2, MERSCoV, and CoV strain 229E in culture fluid. Keywords: SARS-CoV-2, COVID-19, Antibody, Nucleoprotein, Surface plasmon resonance, Immunoassay
1 Department of Materials Science and Engineering, Yonsei University, 50 Yonsei-Ro, Seodaemun-Gu, Seoul 03722, Republic of Korea 2 Molecular Recognition Research Center, Korea Institute of Science and Technology (KIST), Seoul 02792, Republic of Korea 3 Department of Laboratory Medicine, Yonsei University College of Medicine, Seoul 03722, Republic of Korea *Correspondence and requests for materials should be addressed to J.C. Pyun ( [email protected])
Introduction Coronavirus (CoV) has been classified into four genera: alpha, beta, gamma, and delta, with humans being infected by the former two coronaviruses1. CoVs such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), SARS-CoV, and Middle East respiratory syndrome–related coronavirus (MERS-CoV), which are all beta viruses, cause acute respiratory diseases such as coronavirus disease-2019 (COVID19), SARS, and MERS, respectively. These betaCoVs also infect pigs (alpha-CoV and beta-CoV), dogs (alpha-CoV), cats (alpha-CoV), bats (beta-CoV), horses (beta-CoV), and cows (beta-CoV)2. Specifically, the porcine epidemic diarrhea virus (PEDV; alpha-CoV), the transmissible gastroenteritis virus (TGEV; alpha-CoV), and the porcine hemagglutinating encephalomyelitis virus (PHEV; beta-CoV) are known to affect pigs. The CoV genome encodes for four major structural proteins—spike protein, envelope protein, membrane protein, and nucleoprotein (NP). Among these, NP is known to be highly immunogenic and is abundantly expressed during infections3-6. It contains N- and C-terminal binding domains with a linkage region. As a result, it is frequently used in
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