Characterization of novel monoclonal antibodies against MERS-coronavirus spike protein.
Identifieur interne : 000286 ( PubMed/Corpus ); précédent : 000285; suivant : 000287Characterization of novel monoclonal antibodies against MERS-coronavirus spike protein.
Auteurs : Junghyun Goo ; Yuji Jeong ; Young-Shin Park ; Eunji Yang ; Dae-Im Jung ; Semi Rho ; Uni Park ; Hyeyeong Sung ; Pil-Gu Park ; Jung-Ah Choi ; Sang Hwan Seo ; Nam Hyuck Cho ; Hyeja Lee ; Jae Myun Lee ; Jae-Ouk Kim ; Manki SongSource :
- Virus research [ 1872-7492 ] ; 2020.
Abstract
Middle East Respiratory Syndrome coronavirus (MERS-CoV) causes severe pulmonary infection, with ∼35 % mortality. Spike glycoprotein (S) of MERS-CoV is a key target for vaccines and therapeutics because S mediates viral entry and membrane-fusion to host cells. Here, four different S subunit proteins, receptor-binding domain (RBD; 358-606 aa), S1 (1-751 aa), S2 (752-1296 aa), and SΔTM (1-1296 aa), were generated using the baculoviral system and immunized in mice to develop neutralizing antibodies. We developed 77 hybridomas and selected five neutralizing mAbs by immunization with SΔTM against MERS-CoV EMC/2012 strain S-pseudotyped lentivirus. However, all five monoclonal antibodies (mAb) did not neutralize the pseudotyped V534A mutation. Additionally, one mAb RBD-14F8 did not show neutralizing activity against pseudoviruses with amino acid substitution of L506 F or D509 G (England1 strain, EMC/2012 L506 F, and EMC/2012 D509 G), and RBD-43E4 mAb could not neutralize the pseudotyped I529 T mutation, while three other neutralizing mAbs showed broad neutralizing activity. This implies that the mutation in residue 506-509, 529, and 534 of S is critical to generate neutralization escape variants of MERS-CoV. Interestingly, all five neutralizing mAbs have binding affinity to RBD, although most mAbs generated by RBD did not have neutralizing activity. Additionally, chimeric antibodies of RBD-14F8 and RBD-43E4 with human Fc and light chain showed neutralizing effect against wild type MERS-CoV KOR/KNIH/002, similar to the original mouse mAbs. Thus, our mAbs can be utilized for the identification of specific mutations of MERS-CoV.
DOI: 10.1016/j.virusres.2020.197863
PubMed: 31945421
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Electronic address: jokim@ivi.int.</nlm:affiliation></affiliation></author><author><name sortKey="Song, Manki" sort="Song, Manki" uniqKey="Song M" first="Manki" last="Song">Manki Song</name><affiliation><nlm:affiliation>Science Unit, International Vaccine Institute, Seoul, South Korea. Electronic address: mksong@ivi.int.</nlm:affiliation></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2020">2020</date><idno type="RBID">pubmed:31945421</idno><idno type="pmid">31945421</idno><idno type="doi">10.1016/j.virusres.2020.197863</idno><idno type="wicri:Area/PubMed/Corpus">000286</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">000286</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Characterization of novel monoclonal antibodies against MERS-coronavirus spike protein.</title><author><name sortKey="Goo, Junghyun" sort="Goo, Junghyun" uniqKey="Goo J" first="Junghyun" last="Goo">Junghyun Goo</name><affiliation><nlm:affiliation>Science Unit, International Vaccine Institute, Seoul, South Korea.</nlm:affiliation></affiliation></author><author><name sortKey="Jeong, Yuji" sort="Jeong, Yuji" uniqKey="Jeong Y" first="Yuji" last="Jeong">Yuji Jeong</name><affiliation><nlm:affiliation>Science Unit, International Vaccine Institute, Seoul, South Korea.</nlm:affiliation></affiliation></author><author><name sortKey="Park, Young Shin" sort="Park, Young Shin" uniqKey="Park Y" first="Young-Shin" last="Park">Young-Shin Park</name><affiliation><nlm:affiliation>Science Unit, International Vaccine Institute, Seoul, South Korea.</nlm:affiliation></affiliation></author><author><name sortKey="Yang, Eunji" sort="Yang, Eunji" uniqKey="Yang E" first="Eunji" last="Yang">Eunji Yang</name><affiliation><nlm:affiliation>Science Unit, International Vaccine Institute, Seoul, South Korea.</nlm:affiliation></affiliation></author><author><name sortKey="Jung, Dae Im" sort="Jung, Dae Im" uniqKey="Jung D" first="Dae-Im" last="Jung">Dae-Im Jung</name><affiliation><nlm:affiliation>Science Unit, International Vaccine Institute, Seoul, South Korea.</nlm:affiliation></affiliation></author><author><name sortKey="Rho, Semi" sort="Rho, Semi" uniqKey="Rho S" first="Semi" last="Rho">Semi Rho</name><affiliation><nlm:affiliation>Science Unit, International Vaccine Institute, Seoul, South Korea.</nlm:affiliation></affiliation></author><author><name sortKey="Park, Uni" sort="Park, Uni" uniqKey="Park U" first="Uni" last="Park">Uni Park</name><affiliation><nlm:affiliation>Department of Microbiology and Immunology, South Korea; Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul, South Korea.</nlm:affiliation></affiliation></author><author><name sortKey="Sung, Hyeyeong" sort="Sung, Hyeyeong" uniqKey="Sung H" first="Hyeyeong" last="Sung">Hyeyeong Sung</name><affiliation><nlm:affiliation>NKMAX Co., Ltd., Seongnam, South Korea.</nlm:affiliation></affiliation></author><author><name sortKey="Park, Pil Gu" sort="Park, Pil Gu" uniqKey="Park P" first="Pil-Gu" last="Park">Pil-Gu Park</name><affiliation><nlm:affiliation>Department of Microbiology and Immunology, Brain Korea 21 PLUS Project for Medical Science, Institute for Immunology and Immunological Diseases, Yonsei University College of Medicine, Seoul, South Korea.</nlm:affiliation></affiliation></author><author><name sortKey="Choi, Jung Ah" sort="Choi, Jung Ah" uniqKey="Choi J" first="Jung-Ah" last="Choi">Jung-Ah Choi</name><affiliation><nlm:affiliation>Science Unit, International Vaccine Institute, Seoul, South Korea.</nlm:affiliation></affiliation></author><author><name sortKey="Seo, Sang Hwan" sort="Seo, Sang Hwan" uniqKey="Seo S" first="Sang Hwan" last="Seo">Sang Hwan Seo</name><affiliation><nlm:affiliation>Science Unit, International Vaccine Institute, Seoul, South Korea.</nlm:affiliation></affiliation></author><author><name sortKey="Cho, Nam Hyuck" sort="Cho, Nam Hyuck" uniqKey="Cho N" first="Nam Hyuck" last="Cho">Nam Hyuck Cho</name><affiliation><nlm:affiliation>Department of Microbiology and Immunology, South Korea; Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul, South Korea; Institute of Endemic Disease, Seoul National University Medical Research Center and Bundang Hospital, Seoul, South Korea.</nlm:affiliation></affiliation></author><author><name sortKey="Lee, Hyeja" sort="Lee, Hyeja" uniqKey="Lee H" first="Hyeja" last="Lee">Hyeja Lee</name><affiliation><nlm:affiliation>NKMAX Co., Ltd., Seongnam, South Korea.</nlm:affiliation></affiliation></author><author><name sortKey="Lee, Jae Myun" sort="Lee, Jae Myun" uniqKey="Lee J" first="Jae Myun" last="Lee">Jae Myun Lee</name><affiliation><nlm:affiliation>Department of Microbiology and Immunology, Brain Korea 21 PLUS Project for Medical Science, Institute for Immunology and Immunological Diseases, Yonsei University College of Medicine, Seoul, South Korea.</nlm:affiliation></affiliation></author><author><name sortKey="Kim, Jae Ouk" sort="Kim, Jae Ouk" uniqKey="Kim J" first="Jae-Ouk" last="Kim">Jae-Ouk Kim</name><affiliation><nlm:affiliation>Science Unit, International Vaccine Institute, Seoul, South Korea. Electronic address: jokim@ivi.int.</nlm:affiliation></affiliation></author><author><name sortKey="Song, Manki" sort="Song, Manki" uniqKey="Song M" first="Manki" last="Song">Manki Song</name><affiliation><nlm:affiliation>Science Unit, International Vaccine Institute, Seoul, South Korea. Electronic address: mksong@ivi.int.</nlm:affiliation></affiliation></author></analytic><series><title level="j">Virus research</title><idno type="eISSN">1872-7492</idno><imprint><date when="2020" type="published">2020</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Middle East Respiratory Syndrome coronavirus (MERS-CoV) causes severe pulmonary infection, with ∼35 % mortality. Spike glycoprotein (S) of MERS-CoV is a key target for vaccines and therapeutics because S mediates viral entry and membrane-fusion to host cells. Here, four different S subunit proteins, receptor-binding domain (RBD; 358-606 aa), S1 (1-751 aa), S2 (752-1296 aa), and SΔTM (1-1296 aa), were generated using the baculoviral system and immunized in mice to develop neutralizing antibodies. We developed 77 hybridomas and selected five neutralizing mAbs by immunization with SΔTM against MERS-CoV EMC/2012 strain S-pseudotyped lentivirus. However, all five monoclonal antibodies (mAb) did not neutralize the pseudotyped V534A mutation. Additionally, one mAb RBD-14F8 did not show neutralizing activity against pseudoviruses with amino acid substitution of L506 F or D509 G (England1 strain, EMC/2012 L506 F, and EMC/2012 D509 G), and RBD-43E4 mAb could not neutralize the pseudotyped I529 T mutation, while three other neutralizing mAbs showed broad neutralizing activity. This implies that the mutation in residue 506-509, 529, and 534 of S is critical to generate neutralization escape variants of MERS-CoV. Interestingly, all five neutralizing mAbs have binding affinity to RBD, although most mAbs generated by RBD did not have neutralizing activity. Additionally, chimeric antibodies of RBD-14F8 and RBD-43E4 with human Fc and light chain showed neutralizing effect against wild type MERS-CoV KOR/KNIH/002, similar to the original mouse mAbs. Thus, our mAbs can be utilized for the identification of specific mutations of MERS-CoV.</div></front></TEI><pubmed><MedlineCitation Status="In-Data-Review" Owner="NLM"><PMID Version="1">31945421</PMID><DateRevised><Year>2020</Year><Month>04</Month><Day>07</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1872-7492</ISSN><JournalIssue CitedMedium="Internet"><Volume>278</Volume><PubDate><Year>2020</Year><Month>Mar</Month></PubDate></JournalIssue><Title>Virus research</Title><ISOAbbreviation>Virus Res.</ISOAbbreviation></Journal><ArticleTitle>Characterization of novel monoclonal antibodies against MERS-coronavirus spike protein.</ArticleTitle><Pagination><MedlinePgn>197863</MedlinePgn></Pagination><ELocationID EIdType="pii" ValidYN="Y">S0168-1702(19)30750-6</ELocationID><ELocationID EIdType="doi" ValidYN="Y">10.1016/j.virusres.2020.197863</ELocationID><Abstract><AbstractText>Middle East Respiratory Syndrome coronavirus (MERS-CoV) causes severe pulmonary infection, with ∼35 % mortality. Spike glycoprotein (S) of MERS-CoV is a key target for vaccines and therapeutics because S mediates viral entry and membrane-fusion to host cells. Here, four different S subunit proteins, receptor-binding domain (RBD; 358-606 aa), S1 (1-751 aa), S2 (752-1296 aa), and SΔTM (1-1296 aa), were generated using the baculoviral system and immunized in mice to develop neutralizing antibodies. We developed 77 hybridomas and selected five neutralizing mAbs by immunization with SΔTM against MERS-CoV EMC/2012 strain S-pseudotyped lentivirus. However, all five monoclonal antibodies (mAb) did not neutralize the pseudotyped V534A mutation. Additionally, one mAb RBD-14F8 did not show neutralizing activity against pseudoviruses with amino acid substitution of L506 F or D509 G (England1 strain, EMC/2012 L506 F, and EMC/2012 D509 G), and RBD-43E4 mAb could not neutralize the pseudotyped I529 T mutation, while three other neutralizing mAbs showed broad neutralizing activity. This implies that the mutation in residue 506-509, 529, and 534 of S is critical to generate neutralization escape variants of MERS-CoV. Interestingly, all five neutralizing mAbs have binding affinity to RBD, although most mAbs generated by RBD did not have neutralizing activity. Additionally, chimeric antibodies of RBD-14F8 and RBD-43E4 with human Fc and light chain showed neutralizing effect against wild type MERS-CoV KOR/KNIH/002, similar to the original mouse mAbs. Thus, our mAbs can be utilized for the identification of specific mutations of MERS-CoV.</AbstractText><CopyrightInformation>Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Goo</LastName><ForeName>Junghyun</ForeName><Initials>J</Initials><AffiliationInfo><Affiliation>Science Unit, International Vaccine Institute, Seoul, South Korea.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Jeong</LastName><ForeName>Yuji</ForeName><Initials>Y</Initials><AffiliationInfo><Affiliation>Science Unit, International Vaccine Institute, Seoul, South Korea.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Park</LastName><ForeName>Young-Shin</ForeName><Initials>YS</Initials><AffiliationInfo><Affiliation>Science Unit, International Vaccine Institute, Seoul, South Korea.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Yang</LastName><ForeName>Eunji</ForeName><Initials>E</Initials><AffiliationInfo><Affiliation>Science Unit, International Vaccine Institute, Seoul, South Korea.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Jung</LastName><ForeName>Dae-Im</ForeName><Initials>DI</Initials><AffiliationInfo><Affiliation>Science Unit, International Vaccine Institute, Seoul, South Korea.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Rho</LastName><ForeName>Semi</ForeName><Initials>S</Initials><AffiliationInfo><Affiliation>Science Unit, International Vaccine Institute, Seoul, South Korea.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Park</LastName><ForeName>Uni</ForeName><Initials>U</Initials><AffiliationInfo><Affiliation>Department of Microbiology and Immunology, South Korea; Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul, South Korea.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Sung</LastName><ForeName>Hyeyeong</ForeName><Initials>H</Initials><AffiliationInfo><Affiliation>NKMAX Co., Ltd., Seongnam, South Korea.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Park</LastName><ForeName>Pil-Gu</ForeName><Initials>PG</Initials><AffiliationInfo><Affiliation>Department of Microbiology and Immunology, Brain Korea 21 PLUS Project for Medical Science, Institute for Immunology and Immunological Diseases, Yonsei University College of Medicine, Seoul, South Korea.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Choi</LastName><ForeName>Jung-Ah</ForeName><Initials>JA</Initials><AffiliationInfo><Affiliation>Science Unit, International Vaccine Institute, Seoul, South Korea.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Seo</LastName><ForeName>Sang Hwan</ForeName><Initials>SH</Initials><AffiliationInfo><Affiliation>Science Unit, International Vaccine Institute, Seoul, South Korea.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Cho</LastName><ForeName>Nam Hyuck</ForeName><Initials>NH</Initials><AffiliationInfo><Affiliation>Department of Microbiology and Immunology, South Korea; Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul, South Korea; Institute of Endemic Disease, Seoul National University Medical Research Center and Bundang Hospital, Seoul, South Korea.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Lee</LastName><ForeName>Hyeja</ForeName><Initials>H</Initials><AffiliationInfo><Affiliation>NKMAX Co., Ltd., Seongnam, South Korea.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Lee</LastName><ForeName>Jae Myun</ForeName><Initials>JM</Initials><AffiliationInfo><Affiliation>Department of Microbiology and Immunology, Brain Korea 21 PLUS Project for Medical Science, Institute for Immunology and Immunological Diseases, Yonsei University College of Medicine, Seoul, South Korea.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Kim</LastName><ForeName>Jae-Ouk</ForeName><Initials>JO</Initials><AffiliationInfo><Affiliation>Science Unit, International Vaccine Institute, Seoul, South Korea. Electronic address: jokim@ivi.int.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Song</LastName><ForeName>Manki</ForeName><Initials>M</Initials><AffiliationInfo><Affiliation>Science Unit, International Vaccine Institute, Seoul, South Korea. Electronic address: mksong@ivi.int.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2020</Year><Month>01</Month><Day>13</Day></ArticleDate></Article><MedlineJournalInfo><Country>Netherlands</Country><MedlineTA>Virus Res</MedlineTA><NlmUniqueID>8410979</NlmUniqueID><ISSNLinking>0168-1702</ISSNLinking></MedlineJournalInfo><CitationSubset>IM</CitationSubset><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="N">Epitope</Keyword><Keyword MajorTopicYN="N">MERS-CoV</Keyword><Keyword MajorTopicYN="N">Monoclonal antibody</Keyword><Keyword MajorTopicYN="N">Neutralization</Keyword><Keyword MajorTopicYN="N">Neutralizing antibody</Keyword><Keyword MajorTopicYN="N">Pseudovirus</Keyword></KeywordList><CoiStatement>Declaration of Competing Interest The authors declare they have no potential conflicts of interest to disclose.</CoiStatement></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2019</Year><Month>10</Month><Day>18</Day></PubMedPubDate><PubMedPubDate PubStatus="revised"><Year>2020</Year><Month>01</Month><Day>02</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2020</Year><Month>01</Month><Day>10</Day></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2020</Year><Month>1</Month><Day>17</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2020</Year><Month>1</Month><Day>17</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2020</Year><Month>1</Month><Day>17</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">31945421</ArticleId><ArticleId IdType="pii">S0168-1702(19)30750-6</ArticleId><ArticleId IdType="doi">10.1016/j.virusres.2020.197863</ArticleId><ArticleId 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