Selection and Characterization of Monoclonal Antibodies Targeting Middle East Respiratory Syndrome Coronavirus through a Human Synthetic Fab Phage Display Library Panning.
Identifieur interne : 000415 ( PubMed/Curation ); précédent : 000414; suivant : 000416Selection and Characterization of Monoclonal Antibodies Targeting Middle East Respiratory Syndrome Coronavirus through a Human Synthetic Fab Phage Display Library Panning.
Auteurs : Yoonji Kim [Corée du Sud] ; Hansaem Lee [Corée du Sud] ; Keunwan Park [Corée du Sud] ; Sora Park [Corée du Sud] ; Ju-Hyeon Lim [Corée du Sud] ; Min Kyung So [Corée du Sud] ; Hye-Min Woo [Corée du Sud] ; Hyemin Ko [Corée du Sud] ; Jeong-Min Lee [Corée du Sud] ; Sun Hee Lim [Corée du Sud] ; Byoung Joon Ko [Corée du Sud] ; Yeon-Su Park [Corée du Sud] ; So-Young Choi [Corée du Sud] ; Du Hyun Song [Corée du Sud] ; Joo-Yeon Lee [Corée du Sud] ; Sung Soon Kim [Corée du Sud] ; Dae Young Kim [Corée du Sud]Source :
- Antibodies (Basel, Switzerland) [ 2073-4468 ] ; 2019.
Abstract
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 (EC50 = 123-421 nM) for MERS-S2P, showed no cross-reactivity to spike proteins from other CoVs, and were non-aggregating and thermostable (Tm = 61.5-80.4 °C). Reformatting the Fabs into IgGs (Immunoglobulin Gs) greatly increased their apparent affinities (KD = 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 (KD = 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.
DOI: 10.3390/antib8030042
PubMed: 31544848
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<author><name sortKey="So, Min Kyung" sort="So, Min Kyung" uniqKey="So M" first="Min Kyung" last="So">Min Kyung So</name>
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<author><name sortKey="Woo, Hye Min" sort="Woo, Hye Min" uniqKey="Woo H" first="Hye-Min" last="Woo">Hye-Min Woo</name>
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<author><name sortKey="Ko, Hyemin" sort="Ko, Hyemin" uniqKey="Ko H" first="Hyemin" last="Ko">Hyemin Ko</name>
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<author><name sortKey="Song, Du Hyun" sort="Song, Du Hyun" uniqKey="Song D" first="Du Hyun" last="Song">Du Hyun Song</name>
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<author><name sortKey="So, Min Kyung" sort="So, Min Kyung" uniqKey="So M" first="Min Kyung" last="So">Min Kyung So</name>
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<author><name sortKey="Woo, Hye Min" sort="Woo, Hye Min" uniqKey="Woo H" first="Hye-Min" last="Woo">Hye-Min Woo</name>
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<wicri:regionArea>Korea Center for Disease Control, Osong Health Technology Administration Complex, Cheongju-si, Chungcheongbuk-do 28159</wicri:regionArea>
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<author><name sortKey="Ko, Hyemin" sort="Ko, Hyemin" uniqKey="Ko H" first="Hyemin" last="Ko">Hyemin Ko</name>
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<country xml:lang="fr">Corée du Sud</country>
<wicri:regionArea>Korea Center for Disease Control, Osong Health Technology Administration Complex, Cheongju-si, Chungcheongbuk-do 28159</wicri:regionArea>
</affiliation>
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<author><name sortKey="Lee, Jeong Min" sort="Lee, Jeong Min" uniqKey="Lee J" first="Jeong-Min" last="Lee">Jeong-Min Lee</name>
<affiliation wicri:level="1"><nlm:affiliation>Plexense, Inc., Yongin-si, Gyeonggi-do 441-813, Korea.</nlm:affiliation>
<country xml:lang="fr">Corée du Sud</country>
<wicri:regionArea>Plexense, Inc., Yongin-si, Gyeonggi-do 441-813</wicri:regionArea>
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<author><name sortKey="Lim, Sun Hee" sort="Lim, Sun Hee" uniqKey="Lim S" first="Sun Hee" last="Lim">Sun Hee Lim</name>
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<country xml:lang="fr">Corée du Sud</country>
<wicri:regionArea>Plexense, Inc., Yongin-si, Gyeonggi-do 441-813</wicri:regionArea>
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<author><name sortKey="Ko, Byoung Joon" sort="Ko, Byoung Joon" uniqKey="Ko B" first="Byoung Joon" last="Ko">Byoung Joon Ko</name>
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<country xml:lang="fr">Corée du Sud</country>
<wicri:regionArea>New Drug Development Center, Osong Medical Innovation Foundation, Cheongju-si, Chungcheongbuk-do 28160</wicri:regionArea>
</affiliation>
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<author><name sortKey="Park, Yeon Su" sort="Park, Yeon Su" uniqKey="Park Y" first="Yeon-Su" last="Park">Yeon-Su Park</name>
<affiliation wicri:level="1"><nlm:affiliation>Plexense, Inc., Yongin-si, Gyeonggi-do 441-813, Korea.</nlm:affiliation>
<country xml:lang="fr">Corée du Sud</country>
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<author><name sortKey="Choi, So Young" sort="Choi, So Young" uniqKey="Choi S" first="So-Young" last="Choi">So-Young Choi</name>
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<country xml:lang="fr">Corée du Sud</country>
<wicri:regionArea>New Drug Development Center, Osong Medical Innovation Foundation, Cheongju-si, Chungcheongbuk-do 28160</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Song, Du Hyun" sort="Song, Du Hyun" uniqKey="Song D" first="Du Hyun" last="Song">Du Hyun Song</name>
<affiliation wicri:level="1"><nlm:affiliation>New Drug Development Center, Osong Medical Innovation Foundation, Cheongju-si, Chungcheongbuk-do 28160, Korea.</nlm:affiliation>
<country xml:lang="fr">Corée du Sud</country>
<wicri:regionArea>New Drug Development Center, Osong Medical Innovation Foundation, Cheongju-si, Chungcheongbuk-do 28160</wicri:regionArea>
</affiliation>
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<author><name sortKey="Lee, Joo Yeon" sort="Lee, Joo Yeon" uniqKey="Lee J" first="Joo-Yeon" last="Lee">Joo-Yeon Lee</name>
<affiliation wicri:level="1"><nlm:affiliation>Korea Center for Disease Control, Osong Health Technology Administration Complex, Cheongju-si, Chungcheongbuk-do 28159, Korea.</nlm:affiliation>
<country xml:lang="fr">Corée du Sud</country>
<wicri:regionArea>Korea Center for Disease Control, Osong Health Technology Administration Complex, Cheongju-si, Chungcheongbuk-do 28159</wicri:regionArea>
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<author><name sortKey="Kim, Sung Soon" sort="Kim, Sung Soon" uniqKey="Kim S" first="Sung Soon" last="Kim">Sung Soon Kim</name>
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<country xml:lang="fr">Corée du Sud</country>
<wicri:regionArea>Korea Center for Disease Control, Osong Health Technology Administration Complex, Cheongju-si, Chungcheongbuk-do 28159</wicri:regionArea>
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<author><name sortKey="Kim, Dae Young" sort="Kim, Dae Young" uniqKey="Kim D" first="Dae Young" last="Kim">Dae Young Kim</name>
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<country xml:lang="fr">Corée du Sud</country>
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<front><div type="abstract" xml:lang="en">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.</div>
</front>
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<DateRevised><Year>2019</Year>
<Month>10</Month>
<Day>23</Day>
</DateRevised>
<Article PubModel="Electronic"><Journal><ISSN IssnType="Electronic">2073-4468</ISSN>
<JournalIssue CitedMedium="Internet"><Volume>8</Volume>
<Issue>3</Issue>
<PubDate><Year>2019</Year>
<Month>Jul</Month>
<Day>31</Day>
</PubDate>
</JournalIssue>
<Title>Antibodies (Basel, Switzerland)</Title>
<ISOAbbreviation>Antibodies (Basel)</ISOAbbreviation>
</Journal>
<ArticleTitle>Selection and Characterization of Monoclonal Antibodies Targeting Middle East Respiratory Syndrome Coronavirus through a Human Synthetic Fab Phage Display Library Panning.</ArticleTitle>
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<Abstract><AbstractText>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.</AbstractText>
</Abstract>
<AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Kim</LastName>
<ForeName>Yoonji</ForeName>
<Initials>Y</Initials>
<AffiliationInfo><Affiliation>New Drug Development Center, Osong Medical Innovation Foundation, Cheongju-si, Chungcheongbuk-do 28160, Korea.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Lee</LastName>
<ForeName>Hansaem</ForeName>
<Initials>H</Initials>
<AffiliationInfo><Affiliation>Korea Center for Disease Control, Osong Health Technology Administration Complex, Cheongju-si, Chungcheongbuk-do 28159, Korea.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Park</LastName>
<ForeName>Keunwan</ForeName>
<Initials>K</Initials>
<Identifier Source="ORCID">0000-0002-8783-9495</Identifier>
<AffiliationInfo><Affiliation>New Drug Development Center, Osong Medical Innovation Foundation, Cheongju-si, Chungcheongbuk-do 28160, Korea.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Park</LastName>
<ForeName>Sora</ForeName>
<Initials>S</Initials>
<AffiliationInfo><Affiliation>New Drug Development Center, Osong Medical Innovation Foundation, Cheongju-si, Chungcheongbuk-do 28160, Korea.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Lim</LastName>
<ForeName>Ju-Hyeon</ForeName>
<Initials>JH</Initials>
<AffiliationInfo><Affiliation>New Drug Development Center, Osong Medical Innovation Foundation, Cheongju-si, Chungcheongbuk-do 28160, Korea.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>So</LastName>
<ForeName>Min Kyung</ForeName>
<Initials>MK</Initials>
<AffiliationInfo><Affiliation>New Drug Development Center, Osong Medical Innovation Foundation, Cheongju-si, Chungcheongbuk-do 28160, Korea.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Woo</LastName>
<ForeName>Hye-Min</ForeName>
<Initials>HM</Initials>
<AffiliationInfo><Affiliation>Korea Center for Disease Control, Osong Health Technology Administration Complex, Cheongju-si, Chungcheongbuk-do 28159, Korea.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Ko</LastName>
<ForeName>Hyemin</ForeName>
<Initials>H</Initials>
<AffiliationInfo><Affiliation>Korea Center for Disease Control, Osong Health Technology Administration Complex, Cheongju-si, Chungcheongbuk-do 28159, Korea.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Lee</LastName>
<ForeName>Jeong-Min</ForeName>
<Initials>JM</Initials>
<AffiliationInfo><Affiliation>Plexense, Inc., Yongin-si, Gyeonggi-do 441-813, Korea.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Lim</LastName>
<ForeName>Sun Hee</ForeName>
<Initials>SH</Initials>
<AffiliationInfo><Affiliation>Plexense, Inc., Yongin-si, Gyeonggi-do 441-813, Korea.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Ko</LastName>
<ForeName>Byoung Joon</ForeName>
<Initials>BJ</Initials>
<AffiliationInfo><Affiliation>New Drug Development Center, Osong Medical Innovation Foundation, Cheongju-si, Chungcheongbuk-do 28160, Korea.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Park</LastName>
<ForeName>Yeon-Su</ForeName>
<Initials>YS</Initials>
<AffiliationInfo><Affiliation>Plexense, Inc., Yongin-si, Gyeonggi-do 441-813, Korea.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Choi</LastName>
<ForeName>So-Young</ForeName>
<Initials>SY</Initials>
<AffiliationInfo><Affiliation>New Drug Development Center, Osong Medical Innovation Foundation, Cheongju-si, Chungcheongbuk-do 28160, Korea.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Song</LastName>
<ForeName>Du Hyun</ForeName>
<Initials>DH</Initials>
<AffiliationInfo><Affiliation>New Drug Development Center, Osong Medical Innovation Foundation, Cheongju-si, Chungcheongbuk-do 28160, Korea.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Lee</LastName>
<ForeName>Joo-Yeon</ForeName>
<Initials>JY</Initials>
<AffiliationInfo><Affiliation>Korea Center for Disease Control, Osong Health Technology Administration Complex, Cheongju-si, Chungcheongbuk-do 28159, Korea.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Kim</LastName>
<ForeName>Sung Soon</ForeName>
<Initials>SS</Initials>
<AffiliationInfo><Affiliation>Korea Center for Disease Control, Osong Health Technology Administration Complex, Cheongju-si, Chungcheongbuk-do 28159, Korea.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Kim</LastName>
<ForeName>Dae Young</ForeName>
<Initials>DY</Initials>
<AffiliationInfo><Affiliation>New Drug Development Center, Osong Medical Innovation Foundation, Cheongju-si, Chungcheongbuk-do 28160, Korea. kimdae@kbiohealth.kr.</Affiliation>
</AffiliationInfo>
</Author>
</AuthorList>
<Language>eng</Language>
<PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType>
</PublicationTypeList>
<ArticleDate DateType="Electronic"><Year>2019</Year>
<Month>07</Month>
<Day>31</Day>
</ArticleDate>
</Article>
<MedlineJournalInfo><Country>Switzerland</Country>
<MedlineTA>Antibodies (Basel)</MedlineTA>
<NlmUniqueID>101587489</NlmUniqueID>
<ISSNLinking>2073-4468</ISSNLinking>
</MedlineJournalInfo>
<KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="N">MERS-CoV</Keyword>
<Keyword MajorTopicYN="N">S2 subunit</Keyword>
<Keyword MajorTopicYN="N">monoclonal antibody</Keyword>
<Keyword MajorTopicYN="N">phage display</Keyword>
<Keyword MajorTopicYN="N">spike protein</Keyword>
</KeywordList>
</MedlineCitation>
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<Month>06</Month>
<Day>03</Day>
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<PubMedPubDate PubStatus="revised"><Year>2019</Year>
<Month>07</Month>
<Day>01</Day>
</PubMedPubDate>
<PubMedPubDate PubStatus="accepted"><Year>2019</Year>
<Month>07</Month>
<Day>07</Day>
</PubMedPubDate>
<PubMedPubDate PubStatus="entrez"><Year>2019</Year>
<Month>9</Month>
<Day>24</Day>
<Hour>6</Hour>
<Minute>0</Minute>
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<Hour>6</Hour>
<Minute>0</Minute>
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<Minute>1</Minute>
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