Selection and Characterization of Monoclonal Antibodies Targeting Middle East Respiratory Syndrome Coronavirus through a Human Synthetic Fab Phage Display Library Panning
Since its first report in the Middle East in 2012, the Middle East respiratory syndrome-coronavirus (MERS-CoV) has become a global concern due to the high morbidity and mortality of individuals infected with the virus. Although the majority of MERS-CoV cases have been reported in Saudi Arabia, the o...
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MDPI AG
2019-07-01
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Series: | Antibodies |
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Online Access: | https://www.mdpi.com/2073-4468/8/3/42 |
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doaj-fddfb13f0bd54b6d964a98318fb5ef13 |
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record_format |
Article |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Yoonji Kim Hansaem Lee Keunwan Park Sora Park Ju-Hyeon Lim Min Kyung So Hye-Min Woo Hyemin Ko Jeong-Min Lee Sun Hee Lim Byoung Joon Ko Yeon-Su Park So-Young Choi Du Hyun Song Joo-Yeon Lee Sung Soon Kim Dae Young Kim |
spellingShingle |
Yoonji Kim Hansaem Lee Keunwan Park Sora Park Ju-Hyeon Lim Min Kyung So Hye-Min Woo Hyemin Ko Jeong-Min Lee Sun Hee Lim Byoung Joon Ko Yeon-Su Park So-Young Choi Du Hyun Song Joo-Yeon Lee Sung Soon Kim Dae Young Kim Selection and Characterization of Monoclonal Antibodies Targeting Middle East Respiratory Syndrome Coronavirus through a Human Synthetic Fab Phage Display Library Panning Antibodies MERS-CoV spike protein S2 subunit phage display monoclonal antibody |
author_facet |
Yoonji Kim Hansaem Lee Keunwan Park Sora Park Ju-Hyeon Lim Min Kyung So Hye-Min Woo Hyemin Ko Jeong-Min Lee Sun Hee Lim Byoung Joon Ko Yeon-Su Park So-Young Choi Du Hyun Song Joo-Yeon Lee Sung Soon Kim Dae Young Kim |
author_sort |
Yoonji Kim |
title |
Selection and Characterization of Monoclonal Antibodies Targeting Middle East Respiratory Syndrome Coronavirus through a Human Synthetic Fab Phage Display Library Panning |
title_short |
Selection and Characterization of Monoclonal Antibodies Targeting Middle East Respiratory Syndrome Coronavirus through a Human Synthetic Fab Phage Display Library Panning |
title_full |
Selection and Characterization of Monoclonal Antibodies Targeting Middle East Respiratory Syndrome Coronavirus through a Human Synthetic Fab Phage Display Library Panning |
title_fullStr |
Selection and Characterization of Monoclonal Antibodies Targeting Middle East Respiratory Syndrome Coronavirus through a Human Synthetic Fab Phage Display Library Panning |
title_full_unstemmed |
Selection and Characterization of Monoclonal Antibodies Targeting Middle East Respiratory Syndrome Coronavirus through a Human Synthetic Fab Phage Display Library Panning |
title_sort |
selection and characterization of monoclonal antibodies targeting middle east respiratory syndrome coronavirus through a human synthetic fab phage display library panning |
publisher |
MDPI AG |
series |
Antibodies |
issn |
2073-4468 |
publishDate |
2019-07-01 |
description |
Since its first report in the Middle East in 2012, the Middle East respiratory syndrome-coronavirus (MERS-CoV) has become a global concern due to the high morbidity and mortality of individuals infected with the virus. Although the majority of MERS-CoV cases have been reported in Saudi Arabia, the overall risk in areas outside the Middle East remains significant as inside Saudi Arabia. Additional pandemics of MERS-CoV are expected, and thus novel tools and reagents for therapy and diagnosis are urgently needed. Here, we used phage display to develop novel monoclonal antibodies (mAbs) that target MERS-CoV. A human Fab phage display library was panned against the S2 subunit of the MERS-CoV spike protein (MERS-S2P), yielding three unique Fabs (S2A3, S2A6, and S2D5). The Fabs had moderate apparent affinities (Half maximal effective concentration (<i>EC</i><sub>50</sub> = 123−421 nM) for MERS-S2P, showed no cross-reactivity to spike proteins from other CoVs, and were non-aggregating and thermostable (<i>T</i><sub>m</sub> = 61.5−80.4 °C). Reformatting the Fabs into IgGs (Immunoglobulin Gs) greatly increased their apparent affinities (<i>K</i><sub>D</sub> = 0.17−1.2 nM), presumably due to the effects of avidity. These apparent affinities were notably higher than that of a previously reported anti-MERS-CoV S2 reference mAb (<i>K</i><sub>D</sub> = 8.7 nM). Furthermore, two of the three mAbs (S2A3 and S2D5) bound only MERS-CoV (Erasmus Medical Center (EMC)) and not other CoVs, reflecting their high binding specificity. However, the mAbs lacked MERS-CoV neutralizing activity. Given their high affinity, specificity, and desirable stabilities, we anticipate that these anti-MERS-CoV mAbs would be suitable reagents for developing antibody-based diagnostics in laboratory or hospital settings for point-of-care testing. |
topic |
MERS-CoV spike protein S2 subunit phage display monoclonal antibody |
url |
https://www.mdpi.com/2073-4468/8/3/42 |
work_keys_str_mv |
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doaj-fddfb13f0bd54b6d964a98318fb5ef132020-11-25T00:50:12ZengMDPI AGAntibodies2073-44682019-07-01834210.3390/antib8030042antib8030042Selection and Characterization of Monoclonal Antibodies Targeting Middle East Respiratory Syndrome Coronavirus through a Human Synthetic Fab Phage Display Library PanningYoonji Kim0Hansaem Lee1Keunwan Park2Sora Park3Ju-Hyeon Lim4Min Kyung So5Hye-Min Woo6Hyemin Ko7Jeong-Min Lee8Sun Hee Lim9Byoung Joon Ko10Yeon-Su Park11So-Young Choi12Du Hyun Song13Joo-Yeon Lee14Sung Soon Kim15Dae Young Kim16New Drug Development Center, Osong Medical Innovation Foundation, Cheongju-si, Chungcheongbuk-do 28160, KoreaKorea Center for Disease Control, Osong Health Technology Administration Complex, Cheongju-si, Chungcheongbuk-do 28159, KoreaNew Drug Development Center, Osong Medical Innovation Foundation, Cheongju-si, Chungcheongbuk-do 28160, KoreaNew Drug Development Center, Osong Medical Innovation Foundation, Cheongju-si, Chungcheongbuk-do 28160, KoreaNew Drug Development Center, Osong Medical Innovation Foundation, Cheongju-si, Chungcheongbuk-do 28160, KoreaNew Drug Development Center, Osong Medical Innovation Foundation, Cheongju-si, Chungcheongbuk-do 28160, KoreaKorea Center for Disease Control, Osong Health Technology Administration Complex, Cheongju-si, Chungcheongbuk-do 28159, KoreaKorea Center for Disease Control, Osong Health Technology Administration Complex, Cheongju-si, Chungcheongbuk-do 28159, KoreaPlexense, Inc., Yongin-si, Gyeonggi-do 441-813, KoreaPlexense, Inc., Yongin-si, Gyeonggi-do 441-813, KoreaNew Drug Development Center, Osong Medical Innovation Foundation, Cheongju-si, Chungcheongbuk-do 28160, KoreaPlexense, Inc., Yongin-si, Gyeonggi-do 441-813, KoreaNew Drug Development Center, Osong Medical Innovation Foundation, Cheongju-si, Chungcheongbuk-do 28160, KoreaNew Drug Development Center, Osong Medical Innovation Foundation, Cheongju-si, Chungcheongbuk-do 28160, KoreaKorea Center for Disease Control, Osong Health Technology Administration Complex, Cheongju-si, Chungcheongbuk-do 28159, KoreaKorea Center for Disease Control, Osong Health Technology Administration Complex, Cheongju-si, Chungcheongbuk-do 28159, KoreaNew Drug Development Center, Osong Medical Innovation Foundation, Cheongju-si, Chungcheongbuk-do 28160, KoreaSince its first report in the Middle East in 2012, the Middle East respiratory syndrome-coronavirus (MERS-CoV) has become a global concern due to the high morbidity and mortality of individuals infected with the virus. Although the majority of MERS-CoV cases have been reported in Saudi Arabia, the overall risk in areas outside the Middle East remains significant as inside Saudi Arabia. Additional pandemics of MERS-CoV are expected, and thus novel tools and reagents for therapy and diagnosis are urgently needed. Here, we used phage display to develop novel monoclonal antibodies (mAbs) that target MERS-CoV. A human Fab phage display library was panned against the S2 subunit of the MERS-CoV spike protein (MERS-S2P), yielding three unique Fabs (S2A3, S2A6, and S2D5). The Fabs had moderate apparent affinities (Half maximal effective concentration (<i>EC</i><sub>50</sub> = 123−421 nM) for MERS-S2P, showed no cross-reactivity to spike proteins from other CoVs, and were non-aggregating and thermostable (<i>T</i><sub>m</sub> = 61.5−80.4 °C). Reformatting the Fabs into IgGs (Immunoglobulin Gs) greatly increased their apparent affinities (<i>K</i><sub>D</sub> = 0.17−1.2 nM), presumably due to the effects of avidity. These apparent affinities were notably higher than that of a previously reported anti-MERS-CoV S2 reference mAb (<i>K</i><sub>D</sub> = 8.7 nM). Furthermore, two of the three mAbs (S2A3 and S2D5) bound only MERS-CoV (Erasmus Medical Center (EMC)) and not other CoVs, reflecting their high binding specificity. However, the mAbs lacked MERS-CoV neutralizing activity. Given their high affinity, specificity, and desirable stabilities, we anticipate that these anti-MERS-CoV mAbs would be suitable reagents for developing antibody-based diagnostics in laboratory or hospital settings for point-of-care testing.https://www.mdpi.com/2073-4468/8/3/42MERS-CoVspike proteinS2 subunitphage displaymonoclonal antibody |