Engineering a stable CHO cell line for the expression of a MERS-coronavirus vaccine antigen.
Identifieur interne : 000935 ( PubMed/Checkpoint ); précédent : 000934; suivant : 000936Engineering a stable CHO cell line for the expression of a MERS-coronavirus vaccine antigen.
Auteurs : Mun Peak Nyon [États-Unis] ; Lanying Du [États-Unis] ; Chien-Te Kent Tseng [États-Unis] ; Christopher A. Seid [États-Unis] ; Jeroen Pollet [États-Unis] ; Kevin S. Naceanceno [États-Unis] ; Anurodh Agrawal [États-Unis] ; Abdullah Algaissi [États-Unis] ; Bi-Hung Peng [États-Unis] ; Wanbo Tai [États-Unis] ; Shibo Jiang [République populaire de Chine] ; Maria Elena Bottazzi [États-Unis] ; Ulrich Strych [États-Unis] ; Peter J. Hotez [États-Unis]Source :
- Vaccine [ 1873-2518 ] ; 2018.
Descripteurs français
- KwdFr :
- Animaux, Antigènes viraux (génétique), Antigènes viraux (immunologie), Cellules CHO, Coronavirus du syndrome respiratoire du Moyen-Orient (immunologie), Cricetulus, Expression des gènes, Fragments Fc des immunoglobulines (génétique), Fragments Fc des immunoglobulines (immunologie), Génie génétique, Immunogénicité des vaccins, Infections à coronavirus (), Infections à coronavirus (immunologie), Maturation post-traductionnelle des protéines, Protéines recombinantes (génétique), Protéines recombinantes (immunologie), Souris, Vaccins antiviraux (immunologie), Vecteurs génétiques (génétique), Épitopes (), Épitopes (immunologie).
- MESH :
- génétique : Antigènes viraux, Fragments Fc des immunoglobulines, Protéines recombinantes, Vecteurs génétiques.
- immunologie : Antigènes viraux, Coronavirus du syndrome respiratoire du Moyen-Orient, Fragments Fc des immunoglobulines, Infections à coronavirus, Protéines recombinantes, Vaccins antiviraux, Épitopes.
- Animaux, Cellules CHO, Cricetulus, Expression des gènes, Génie génétique, Immunogénicité des vaccins, Infections à coronavirus, Maturation post-traductionnelle des protéines, Souris, Épitopes.
English descriptors
- KwdEn :
- Animals, Antigens, Viral (genetics), Antigens, Viral (immunology), CHO Cells, Coronavirus Infections (immunology), Coronavirus Infections (prevention & control), Cricetulus, Epitopes (chemistry), Epitopes (immunology), Gene Expression, Genetic Engineering, Genetic Vectors (genetics), Immunogenicity, Vaccine, Immunoglobulin Fc Fragments (genetics), Immunoglobulin Fc Fragments (immunology), Mice, Middle East Respiratory Syndrome Coronavirus (immunology), Protein Processing, Post-Translational, Recombinant Proteins (genetics), Recombinant Proteins (immunology), Viral Vaccines (immunology).
- MESH :
- chemical , chemistry : Epitopes.
- chemical , genetics : Antigens, Viral, Immunoglobulin Fc Fragments, Recombinant Proteins.
- chemical , immunology : Antigens, Viral, Epitopes, Immunoglobulin Fc Fragments, Recombinant Proteins, Viral Vaccines.
- genetics : Genetic Vectors.
- immunology : Coronavirus Infections, Middle East Respiratory Syndrome Coronavirus.
- prevention & control : Coronavirus Infections.
- Animals, CHO Cells, Cricetulus, Gene Expression, Genetic Engineering, Immunogenicity, Vaccine, Mice, Protein Processing, Post-Translational.
Abstract
Middle East respiratory syndrome coronavirus (MERS-CoV) has infected at least 2040 patients and caused 712 deaths since its first appearance in 2012, yet neither pathogen-specific therapeutics nor approved vaccines are available. To address this need, we are developing a subunit recombinant protein vaccine comprising residues 377-588 of the MERS-CoV spike protein receptor-binding domain (RBD), which, when formulated with the AddaVax adjuvant, it induces a significant neutralizing antibody response and protection against MERS-CoV challenge in vaccinated animals. To prepare for the manufacture and first-in-human testing of the vaccine, we have developed a process to stably produce the recombinant MERS S377-588 protein in Chinese hamster ovary (CHO) cells. To accomplish this, we transfected an adherent dihydrofolate reductase-deficient CHO cell line (adCHO) with a plasmid encoding S377-588 fused with the human IgG Fc fragment (S377-588-Fc). We then demonstrated the interleukin-2 signal peptide-directed secretion of the recombinant protein into extracellular milieu. Using a gradually increasing methotrexate (MTX) concentration to 5 μM, we increased protein yield by a factor of 40. The adCHO-expressed S377-588-Fc recombinant protein demonstrated functionality and binding specificity identical to those of the protein from transiently transfected HEK293T cells. In addition, hCD26/dipeptidyl peptidase-4 (DPP4) transgenic mice vaccinated with AddaVax-adjuvanted S377-588-Fc could produce neutralizing antibodies against MERS-CoV and survived for at least 21 days after challenge with live MERS-CoV with no evidence of immunological toxicity or eosinophilic immune enhancement. To prepare for large scale-manufacture of the vaccine antigen, we have further developed a high-yield monoclonal suspension CHO cell line.
DOI: 10.1016/j.vaccine.2018.02.065
PubMed: 29496347
Affiliations:
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Kimball Research Institute, New York Blood Center, New York, NY, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Lindsley F. Kimball Research Institute, New York Blood Center, New York, NY</wicri:regionArea><placeName><region type="state">État de New York</region></placeName></affiliation></author><author><name sortKey="Jiang, Shibo" sort="Jiang, Shibo" uniqKey="Jiang S" first="Shibo" last="Jiang">Shibo Jiang</name><affiliation wicri:level="1"><nlm:affiliation>Lindsley F. Kimball Research Institute, New York Blood Center, New York, NY, USA; Key Laboratory of Medical Molecular Virology of MOE/MOH, School of Basic Medical Sciences, Fudan University, Shanghai, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Lindsley F. Kimball Research Institute, New York Blood Center, New York, NY, USA; Key Laboratory of Medical Molecular Virology of MOE/MOH, School of Basic Medical Sciences, Fudan University, Shanghai</wicri:regionArea><wicri:noRegion>Shanghai</wicri:noRegion></affiliation></author><author><name sortKey="Bottazzi, Maria Elena" sort="Bottazzi, Maria Elena" uniqKey="Bottazzi M" first="Maria Elena" last="Bottazzi">Maria Elena Bottazzi</name><affiliation wicri:level="2"><nlm:affiliation>Texas Children's Hospital Center for Vaccine Development, USA; Department of Pediatrics, National School of Tropical Medicine, Baylor College of Medicine, Houston, TX, USA. 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Kimball Research Institute, New York Blood Center, New York, NY, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Lindsley F. Kimball Research Institute, New York Blood Center, New York, NY</wicri:regionArea><placeName><region type="state">État de New York</region></placeName></affiliation></author><author><name sortKey="Tseng, Chien Te Kent" sort="Tseng, Chien Te Kent" uniqKey="Tseng C" first="Chien-Te Kent" last="Tseng">Chien-Te Kent Tseng</name><affiliation wicri:level="2"><nlm:affiliation>Department of Microbiology and Immunology & Center of Biodefense and Emerging Diseases, University of Texas Medical Branch, Galveston, TX, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Microbiology and Immunology & Center of Biodefense and Emerging Diseases, University of Texas Medical Branch, Galveston, TX</wicri:regionArea><placeName><region type="state">Texas</region></placeName></affiliation></author><author><name sortKey="Seid, Christopher A" sort="Seid, Christopher A" uniqKey="Seid C" first="Christopher A" last="Seid">Christopher A. Seid</name><affiliation wicri:level="2"><nlm:affiliation>Texas Children's Hospital Center for Vaccine Development, USA; Department of Pediatrics, National School of Tropical Medicine, Baylor College of Medicine, Houston, TX, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Texas Children's Hospital Center for Vaccine Development, USA; Department of Pediatrics, National School of Tropical Medicine, Baylor College of Medicine, Houston, TX</wicri:regionArea><placeName><region type="state">Texas</region></placeName></affiliation></author><author><name sortKey="Pollet, Jeroen" sort="Pollet, Jeroen" uniqKey="Pollet J" first="Jeroen" last="Pollet">Jeroen Pollet</name><affiliation wicri:level="2"><nlm:affiliation>Texas Children's Hospital Center for Vaccine Development, USA; Department of Pediatrics, National School of Tropical Medicine, Baylor College of Medicine, Houston, TX, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Texas Children's Hospital Center for Vaccine Development, USA; Department of Pediatrics, National School of Tropical Medicine, Baylor College of Medicine, Houston, TX</wicri:regionArea><placeName><region type="state">Texas</region></placeName></affiliation></author><author><name sortKey="Naceanceno, Kevin S" sort="Naceanceno, Kevin S" uniqKey="Naceanceno K" first="Kevin S" last="Naceanceno">Kevin S. Naceanceno</name><affiliation wicri:level="2"><nlm:affiliation>Texas Children's Hospital Center for Vaccine Development, USA; Department of Pediatrics, National School of Tropical Medicine, Baylor College of Medicine, Houston, TX, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Texas Children's Hospital Center for Vaccine Development, USA; Department of Pediatrics, National School of Tropical Medicine, Baylor College of Medicine, Houston, TX</wicri:regionArea><placeName><region type="state">Texas</region></placeName></affiliation></author><author><name sortKey="Agrawal, Anurodh" sort="Agrawal, Anurodh" uniqKey="Agrawal A" first="Anurodh" last="Agrawal">Anurodh Agrawal</name><affiliation wicri:level="2"><nlm:affiliation>Department of Microbiology and Immunology & Center of Biodefense and Emerging Diseases, University of Texas Medical Branch, Galveston, TX, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Microbiology and Immunology & Center of Biodefense and Emerging Diseases, University of Texas Medical Branch, Galveston, TX</wicri:regionArea><placeName><region type="state">Texas</region></placeName></affiliation></author><author><name sortKey="Algaissi, Abdullah" sort="Algaissi, Abdullah" uniqKey="Algaissi A" first="Abdullah" last="Algaissi">Abdullah Algaissi</name><affiliation wicri:level="2"><nlm:affiliation>Department of Microbiology and Immunology & Center of Biodefense and Emerging Diseases, University of Texas Medical Branch, Galveston, TX, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Microbiology and Immunology & Center of Biodefense and Emerging Diseases, University of Texas Medical Branch, Galveston, TX</wicri:regionArea><placeName><region type="state">Texas</region></placeName></affiliation></author><author><name sortKey="Peng, Bi Hung" sort="Peng, Bi Hung" uniqKey="Peng B" first="Bi-Hung" last="Peng">Bi-Hung Peng</name><affiliation wicri:level="2"><nlm:affiliation>Department of Microbiology and Immunology & Center of Biodefense and Emerging Diseases, University of Texas Medical Branch, Galveston, TX, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Microbiology and Immunology & Center of Biodefense and Emerging Diseases, University of Texas Medical Branch, Galveston, TX</wicri:regionArea><placeName><region type="state">Texas</region></placeName></affiliation></author><author><name sortKey="Tai, Wanbo" sort="Tai, Wanbo" uniqKey="Tai W" first="Wanbo" last="Tai">Wanbo Tai</name><affiliation wicri:level="2"><nlm:affiliation>Lindsley F. Kimball Research Institute, New York Blood Center, New York, NY, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Lindsley F. Kimball Research Institute, New York Blood Center, New York, NY</wicri:regionArea><placeName><region type="state">État de New York</region></placeName></affiliation></author><author><name sortKey="Jiang, Shibo" sort="Jiang, Shibo" uniqKey="Jiang S" first="Shibo" last="Jiang">Shibo Jiang</name><affiliation wicri:level="1"><nlm:affiliation>Lindsley F. Kimball Research Institute, New York Blood Center, New York, NY, USA; Key Laboratory of Medical Molecular Virology of MOE/MOH, School of Basic Medical Sciences, Fudan University, Shanghai, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Lindsley F. Kimball Research Institute, New York Blood Center, New York, NY, USA; Key Laboratory of Medical Molecular Virology of MOE/MOH, School of Basic Medical Sciences, Fudan University, Shanghai</wicri:regionArea><wicri:noRegion>Shanghai</wicri:noRegion></affiliation></author><author><name sortKey="Bottazzi, Maria Elena" sort="Bottazzi, Maria Elena" uniqKey="Bottazzi M" first="Maria Elena" last="Bottazzi">Maria Elena Bottazzi</name><affiliation wicri:level="2"><nlm:affiliation>Texas Children's Hospital Center for Vaccine Development, USA; Department of Pediatrics, National School of Tropical Medicine, Baylor College of Medicine, Houston, TX, USA. Electronic address: bottazzi@bcm.edu.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Texas Children's Hospital Center for Vaccine Development, USA; Department of Pediatrics, National School of Tropical Medicine, Baylor College of Medicine, Houston, TX</wicri:regionArea><placeName><region type="state">Texas</region></placeName></affiliation></author><author><name sortKey="Strych, Ulrich" sort="Strych, Ulrich" uniqKey="Strych U" first="Ulrich" last="Strych">Ulrich Strych</name><affiliation wicri:level="2"><nlm:affiliation>Texas Children's Hospital Center for Vaccine Development, USA; Department of Pediatrics, National School of Tropical Medicine, Baylor College of Medicine, Houston, TX, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Texas Children's Hospital Center for Vaccine Development, USA; Department of Pediatrics, National School of Tropical Medicine, Baylor College of Medicine, Houston, TX</wicri:regionArea><placeName><region type="state">Texas</region></placeName></affiliation></author><author><name sortKey="Hotez, Peter J" sort="Hotez, Peter J" uniqKey="Hotez P" first="Peter J" last="Hotez">Peter J. Hotez</name><affiliation wicri:level="2"><nlm:affiliation>Texas Children's Hospital Center for Vaccine Development, USA; Department of Pediatrics, National School of Tropical Medicine, Baylor College of Medicine, Houston, TX, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Texas Children's Hospital Center for Vaccine Development, USA; Department of Pediatrics, National School of Tropical Medicine, Baylor College of Medicine, Houston, TX</wicri:regionArea><placeName><region type="state">Texas</region></placeName></affiliation></author></analytic><series><title level="j">Vaccine</title><idno type="eISSN">1873-2518</idno><imprint><date when="2018" type="published">2018</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Antigens, Viral (genetics)</term><term>Antigens, Viral (immunology)</term><term>CHO Cells</term><term>Coronavirus Infections (immunology)</term><term>Coronavirus Infections (prevention & control)</term><term>Cricetulus</term><term>Epitopes (chemistry)</term><term>Epitopes (immunology)</term><term>Gene Expression</term><term>Genetic Engineering</term><term>Genetic Vectors (genetics)</term><term>Immunogenicity, Vaccine</term><term>Immunoglobulin Fc Fragments (genetics)</term><term>Immunoglobulin Fc Fragments (immunology)</term><term>Mice</term><term>Middle East Respiratory Syndrome Coronavirus (immunology)</term><term>Protein Processing, Post-Translational</term><term>Recombinant Proteins (genetics)</term><term>Recombinant Proteins (immunology)</term><term>Viral Vaccines (immunology)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Animaux</term><term>Antigènes viraux (génétique)</term><term>Antigènes viraux (immunologie)</term><term>Cellules CHO</term><term>Coronavirus du syndrome respiratoire du Moyen-Orient (immunologie)</term><term>Cricetulus</term><term>Expression des gènes</term><term>Fragments Fc des immunoglobulines (génétique)</term><term>Fragments Fc des immunoglobulines (immunologie)</term><term>Génie génétique</term><term>Immunogénicité des vaccins</term><term>Infections à coronavirus ()</term><term>Infections à coronavirus (immunologie)</term><term>Maturation post-traductionnelle des protéines</term><term>Protéines recombinantes (génétique)</term><term>Protéines recombinantes (immunologie)</term><term>Souris</term><term>Vaccins antiviraux (immunologie)</term><term>Vecteurs génétiques (génétique)</term><term>Épitopes ()</term><term>Épitopes (immunologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Epitopes</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Antigens, Viral</term><term>Immunoglobulin Fc Fragments</term><term>Recombinant Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Antigens, Viral</term><term>Epitopes</term><term>Immunoglobulin Fc Fragments</term><term>Recombinant Proteins</term><term>Viral Vaccines</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Genetic Vectors</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Antigènes viraux</term><term>Fragments Fc des immunoglobulines</term><term>Protéines recombinantes</term><term>Vecteurs génétiques</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Antigènes viraux</term><term>Coronavirus du syndrome respiratoire du Moyen-Orient</term><term>Fragments Fc des immunoglobulines</term><term>Infections à coronavirus</term><term>Protéines recombinantes</term><term>Vaccins antiviraux</term><term>Épitopes</term></keywords><keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>Coronavirus Infections</term><term>Middle East Respiratory Syndrome Coronavirus</term></keywords><keywords scheme="MESH" qualifier="prevention & control" xml:lang="en"><term>Coronavirus Infections</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>CHO Cells</term><term>Cricetulus</term><term>Gene Expression</term><term>Genetic Engineering</term><term>Immunogenicity, Vaccine</term><term>Mice</term><term>Protein Processing, Post-Translational</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Animaux</term><term>Cellules CHO</term><term>Cricetulus</term><term>Expression des gènes</term><term>Génie génétique</term><term>Immunogénicité des vaccins</term><term>Infections à coronavirus</term><term>Maturation post-traductionnelle des protéines</term><term>Souris</term><term>Épitopes</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Middle East respiratory syndrome coronavirus (MERS-CoV) has infected at least 2040 patients and caused 712 deaths since its first appearance in 2012, yet neither pathogen-specific therapeutics nor approved vaccines are available. To address this need, we are developing a subunit recombinant protein vaccine comprising residues 377-588 of the MERS-CoV spike protein receptor-binding domain (RBD), which, when formulated with the AddaVax adjuvant, it induces a significant neutralizing antibody response and protection against MERS-CoV challenge in vaccinated animals. To prepare for the manufacture and first-in-human testing of the vaccine, we have developed a process to stably produce the recombinant MERS S377-588 protein in Chinese hamster ovary (CHO) cells. To accomplish this, we transfected an adherent dihydrofolate reductase-deficient CHO cell line (adCHO) with a plasmid encoding S377-588 fused with the human IgG Fc fragment (S377-588-Fc). We then demonstrated the interleukin-2 signal peptide-directed secretion of the recombinant protein into extracellular milieu. Using a gradually increasing methotrexate (MTX) concentration to 5 μM, we increased protein yield by a factor of 40. The adCHO-expressed S377-588-Fc recombinant protein demonstrated functionality and binding specificity identical to those of the protein from transiently transfected HEK293T cells. In addition, hCD26/dipeptidyl peptidase-4 (DPP4) transgenic mice vaccinated with AddaVax-adjuvanted S377-588-Fc could produce neutralizing antibodies against MERS-CoV and survived for at least 21 days after challenge with live MERS-CoV with no evidence of immunological toxicity or eosinophilic immune enhancement. To prepare for large scale-manufacture of the vaccine antigen, we have further developed a high-yield monoclonal suspension CHO cell line.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">29496347</PMID><DateCompleted><Year>2018</Year><Month>09</Month><Day>11</Day></DateCompleted><DateRevised><Year>2020</Year><Month>04</Month><Day>07</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1873-2518</ISSN><JournalIssue CitedMedium="Internet"><Volume>36</Volume><Issue>14</Issue><PubDate><Year>2018</Year><Month>03</Month><Day>27</Day></PubDate></JournalIssue><Title>Vaccine</Title><ISOAbbreviation>Vaccine</ISOAbbreviation></Journal><ArticleTitle>Engineering a stable CHO cell line for the expression of a MERS-coronavirus vaccine antigen.</ArticleTitle><Pagination><MedlinePgn>1853-1862</MedlinePgn></Pagination><ELocationID EIdType="pii" ValidYN="Y">S0264-410X(18)30252-4</ELocationID><ELocationID EIdType="doi" ValidYN="Y">10.1016/j.vaccine.2018.02.065</ELocationID><Abstract><AbstractText>Middle East respiratory syndrome coronavirus (MERS-CoV) has infected at least 2040 patients and caused 712 deaths since its first appearance in 2012, yet neither pathogen-specific therapeutics nor approved vaccines are available. To address this need, we are developing a subunit recombinant protein vaccine comprising residues 377-588 of the MERS-CoV spike protein receptor-binding domain (RBD), which, when formulated with the AddaVax adjuvant, it induces a significant neutralizing antibody response and protection against MERS-CoV challenge in vaccinated animals. To prepare for the manufacture and first-in-human testing of the vaccine, we have developed a process to stably produce the recombinant MERS S377-588 protein in Chinese hamster ovary (CHO) cells. To accomplish this, we transfected an adherent dihydrofolate reductase-deficient CHO cell line (adCHO) with a plasmid encoding S377-588 fused with the human IgG Fc fragment (S377-588-Fc). We then demonstrated the interleukin-2 signal peptide-directed secretion of the recombinant protein into extracellular milieu. Using a gradually increasing methotrexate (MTX) concentration to 5 μM, we increased protein yield by a factor of 40. The adCHO-expressed S377-588-Fc recombinant protein demonstrated functionality and binding specificity identical to those of the protein from transiently transfected HEK293T cells. In addition, hCD26/dipeptidyl peptidase-4 (DPP4) transgenic mice vaccinated with AddaVax-adjuvanted S377-588-Fc could produce neutralizing antibodies against MERS-CoV and survived for at least 21 days after challenge with live MERS-CoV with no evidence of immunological toxicity or eosinophilic immune enhancement. To prepare for large scale-manufacture of the vaccine antigen, we have further developed a high-yield monoclonal suspension CHO cell line.</AbstractText><CopyrightInformation>Copyright © 2018 Elsevier Ltd. All rights reserved.</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Nyon</LastName><ForeName>Mun Peak</ForeName><Initials>MP</Initials><AffiliationInfo><Affiliation>Tropical Infectious Diseases Research and Education Centre (TIDREC), University of Malaya, Kuala Lumpur, Malaysia; Texas Children's Hospital Center for Vaccine Development, USA; Department of Pediatrics, National School of Tropical Medicine, Baylor College of Medicine, Houston, TX, USA.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Du</LastName><ForeName>Lanying</ForeName><Initials>L</Initials><AffiliationInfo><Affiliation>Lindsley F. Kimball Research Institute, New York Blood Center, New York, NY, USA.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Tseng</LastName><ForeName>Chien-Te Kent</ForeName><Initials>CK</Initials><AffiliationInfo><Affiliation>Department of Microbiology and Immunology & Center of Biodefense and Emerging Diseases, University of Texas Medical Branch, Galveston, TX, USA.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Seid</LastName><ForeName>Christopher A</ForeName><Initials>CA</Initials><AffiliationInfo><Affiliation>Texas Children's Hospital Center for Vaccine Development, USA; Department of Pediatrics, National School of Tropical Medicine, Baylor College of Medicine, Houston, TX, USA.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Pollet</LastName><ForeName>Jeroen</ForeName><Initials>J</Initials><AffiliationInfo><Affiliation>Texas Children's Hospital Center for Vaccine Development, USA; Department of Pediatrics, National School of Tropical Medicine, Baylor College of Medicine, Houston, TX, USA.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Naceanceno</LastName><ForeName>Kevin S</ForeName><Initials>KS</Initials><AffiliationInfo><Affiliation>Texas Children's Hospital Center for Vaccine Development, USA; Department of Pediatrics, National School of Tropical Medicine, Baylor College of Medicine, Houston, TX, USA.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Agrawal</LastName><ForeName>Anurodh</ForeName><Initials>A</Initials><AffiliationInfo><Affiliation>Department of Microbiology and Immunology & Center of Biodefense and Emerging Diseases, University of Texas Medical Branch, Galveston, TX, USA.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Algaissi</LastName><ForeName>Abdullah</ForeName><Initials>A</Initials><AffiliationInfo><Affiliation>Department of Microbiology and Immunology & Center of Biodefense and Emerging Diseases, University of Texas Medical Branch, Galveston, TX, USA.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Peng</LastName><ForeName>Bi-Hung</ForeName><Initials>BH</Initials><AffiliationInfo><Affiliation>Department of Microbiology and Immunology & Center of Biodefense and Emerging Diseases, University of Texas Medical Branch, Galveston, TX, USA.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Tai</LastName><ForeName>Wanbo</ForeName><Initials>W</Initials><AffiliationInfo><Affiliation>Lindsley F. Kimball Research Institute, New York Blood Center, New York, NY, USA.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Jiang</LastName><ForeName>Shibo</ForeName><Initials>S</Initials><AffiliationInfo><Affiliation>Lindsley F. Kimball Research Institute, New York Blood Center, New York, NY, USA; Key Laboratory of Medical Molecular Virology of MOE/MOH, School of Basic Medical Sciences, Fudan University, Shanghai, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Bottazzi</LastName><ForeName>Maria Elena</ForeName><Initials>ME</Initials><AffiliationInfo><Affiliation>Texas Children's Hospital Center for Vaccine Development, USA; Department of Pediatrics, National School of Tropical Medicine, Baylor College of Medicine, Houston, TX, USA. Electronic address: bottazzi@bcm.edu.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Strych</LastName><ForeName>Ulrich</ForeName><Initials>U</Initials><AffiliationInfo><Affiliation>Texas Children's Hospital Center for Vaccine Development, USA; Department of Pediatrics, National School of Tropical Medicine, Baylor College of Medicine, Houston, TX, USA.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Hotez</LastName><ForeName>Peter J</ForeName><Initials>PJ</Initials><AffiliationInfo><Affiliation>Texas Children's Hospital Center for Vaccine Development, USA; Department of Pediatrics, National School of Tropical Medicine, Baylor College of Medicine, Houston, TX, USA.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><GrantList CompleteYN="Y"><Grant><GrantID>R01 AI098775</GrantID><Acronym>AI</Acronym><Agency>NIAID NIH HHS</Agency><Country>United States</Country></Grant><Grant><GrantID>R21 AI128311</GrantID><Acronym>AI</Acronym><Agency>NIAID NIH HHS</Agency><Country>United States</Country></Grant></GrantList><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D052061">Research Support, N.I.H., Extramural</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2018</Year><Month>02</Month><Day>26</Day></ArticleDate></Article><MedlineJournalInfo><Country>Netherlands</Country><MedlineTA>Vaccine</MedlineTA><NlmUniqueID>8406899</NlmUniqueID><ISSNLinking>0264-410X</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D000956">Antigens, Viral</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D000939">Epitopes</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D007141">Immunoglobulin Fc Fragments</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D011994">Recombinant Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D014765">Viral Vaccines</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D000818" MajorTopicYN="N">Animals</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D000956" MajorTopicYN="N">Antigens, Viral</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName><QualifierName UI="Q000276" MajorTopicYN="Y">immunology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D016466" MajorTopicYN="N">CHO Cells</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D018352" MajorTopicYN="N">Coronavirus Infections</DescriptorName><QualifierName UI="Q000276" MajorTopicYN="Y">immunology</QualifierName><QualifierName UI="Q000517" 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Fragments</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000276" MajorTopicYN="N">immunology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D051379" MajorTopicYN="N">Mice</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D065207" MajorTopicYN="N">Middle East Respiratory Syndrome Coronavirus</DescriptorName><QualifierName UI="Q000276" MajorTopicYN="Y">immunology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D011499" MajorTopicYN="N">Protein Processing, Post-Translational</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D011994" MajorTopicYN="N">Recombinant Proteins</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000276" MajorTopicYN="N">immunology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D014765" MajorTopicYN="N">Viral Vaccines</DescriptorName><QualifierName UI="Q000276" MajorTopicYN="Y">immunology</QualifierName></MeshHeading></MeshHeadingList><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="Y">Chinese hamster ovary cells</Keyword><Keyword MajorTopicYN="Y">Middle East respiratory syndrome coronavirus</Keyword><Keyword MajorTopicYN="Y">Receptor binding domain</Keyword></KeywordList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2017</Year><Month>10</Month><Day>12</Day></PubMedPubDate><PubMedPubDate PubStatus="revised"><Year>2018</Year><Month>02</Month><Day>12</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2018</Year><Month>02</Month><Day>16</Day></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2018</Year><Month>3</Month><Day>3</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2018</Year><Month>9</Month><Day>12</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2018</Year><Month>3</Month><Day>3</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">29496347</ArticleId><ArticleId IdType="pii">S0264-410X(18)30252-4</ArticleId><ArticleId IdType="doi">10.1016/j.vaccine.2018.02.065</ArticleId><ArticleId IdType="pmc">PMC5860679</ArticleId><ArticleId IdType="mid">NIHMS947559</ArticleId></ArticleIdList><ReferenceList><Reference><Citation>Adv Pharm Bull. 2013;3(2):257-63</Citation><ArticleIdList><ArticleId IdType="pubmed">24312845</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>EMBO Mol Med. 2012 Oct;4(10):1015-28</Citation><ArticleIdList><ArticleId IdType="pubmed">22837174</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>J Virol. 2016 Dec 16;91(1):</Citation><ArticleIdList><ArticleId IdType="pubmed">27795425</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>J Virol. 2015 Feb;89(4):1954-64</Citation><ArticleIdList><ArticleId IdType="pubmed">25428871</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Cell Mol Immunol. 2016 Mar;13(2):180-90</Citation><ArticleIdList><ArticleId IdType="pubmed">25640653</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Expert Rev Vaccines. 2016 Sep;15(9):1123-34</Citation><ArticleIdList><ArticleId IdType="pubmed">26985862</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>J Virol. 2014 Jun;88(12):7045-53</Citation><ArticleIdList><ArticleId IdType="pubmed">24719424</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Sleep Med. 2014 Jun;15(6):654-60</Citation><ArticleIdList><ArticleId IdType="pubmed">24831248</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Expert Opin Ther Targets. 2017 Feb;21(2):131-143</Citation><ArticleIdList><ArticleId IdType="pubmed">27936982</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Virus Res. 2015 Apr 16;202:151-9</Citation><ArticleIdList><ArticleId IdType="pubmed">25445336</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Vaccine. 2014 Oct 21;32(46):6170-6176</Citation><ArticleIdList><ArticleId IdType="pubmed">25240756</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>J Biotechnol. 2012 Dec 31;162(2-3):171-82</Citation><ArticleIdList><ArticleId IdType="pubmed">22981627</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Appl Microbiol Biotechnol. 2015 Dec;99(24):10467-80</Citation><ArticleIdList><ArticleId IdType="pubmed">26373723</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>PLoS One. 2012;7(1):e30083</Citation><ArticleIdList><ArticleId IdType="pubmed">22272277</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Adv Exp Med Biol. 2017;972:49-60</Citation><ArticleIdList><ArticleId IdType="pubmed">27966107</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Cytotechnology. 2000 Jul;33(1-3):37-46</Citation><ArticleIdList><ArticleId IdType="pubmed">19002809</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Vaccine. 2011 Apr 5;29(16):2968-77</Citation><ArticleIdList><ArticleId IdType="pubmed">21329775</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>PLoS One. 2013 Dec 04;8(12):e81587</Citation><ArticleIdList><ArticleId IdType="pubmed">24324708</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Nat Commun. 2015 Sep 15;6:8223</Citation><ArticleIdList><ArticleId IdType="pubmed">26370782</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Expert Opin Biol Ther. 2015;15(11):1647-51</Citation><ArticleIdList><ArticleId IdType="pubmed">26414077</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>J Virol. 2015 Apr;89(7):3659-70</Citation><ArticleIdList><ArticleId IdType="pubmed">25589660</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Biotechnol Bioeng. 2013 Apr;110(4):1164-73</Citation><ArticleIdList><ArticleId IdType="pubmed">23124363</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>J Virol. 2011 Oct;85(20):10542-53</Citation><ArticleIdList><ArticleId IdType="pubmed">21849464</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>J Immunol Methods. 1992 Jul 31;152(1):89-104</Citation><ArticleIdList><ArticleId IdType="pubmed">1640112</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>J Virol. 2014 Jul;88(14):7796-805</Citation><ArticleIdList><ArticleId IdType="pubmed">24789777</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>J Virol. 2015 Oct 07;90(1):57-67</Citation><ArticleIdList><ArticleId IdType="pubmed">26446606</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Vaccines (Basel). 2013 Jul 18;1(3):278-92</Citation><ArticleIdList><ArticleId IdType="pubmed">26344113</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>J Virol. 2013 Sep;87(17):9939-42</Citation><ArticleIdList><ArticleId IdType="pubmed">23824801</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Pharmaceuticals (Basel). 2013 Apr 26;6(5):579-603</Citation><ArticleIdList><ArticleId IdType="pubmed">24276168</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>J Virol. 2011 Mar;85(6):3010-4</Citation><ArticleIdList><ArticleId IdType="pubmed">21191017</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Hum Vaccin Immunother. 2017 Jul 3;13(7):1615-1624</Citation><ArticleIdList><ArticleId IdType="pubmed">28277821</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Hum Vaccin Immunother. 2014;10(3):648-58</Citation><ArticleIdList><ArticleId IdType="pubmed">24355931</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Expert Rev Vaccines. 2012 Dec;11(12):1405-13</Citation><ArticleIdList><ArticleId IdType="pubmed">23252385</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Nature. 1989 Feb 9;337(6207):525-31</Citation><ArticleIdList><ArticleId IdType="pubmed">2536900</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>J Infect Public Health. 2017 Mar - Apr;10(2):191-194</Citation><ArticleIdList><ArticleId IdType="pubmed">27140697</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Cytotechnology. 2013 Oct;65(5):791-3</Citation><ArticleIdList><ArticleId IdType="pubmed">23975256</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Virology. 2016 Dec;499:375-382</Citation><ArticleIdList><ArticleId IdType="pubmed">27750111</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Hum Vaccin Immunother. 2016 Sep;12(9):2351-6</Citation><ArticleIdList><ArticleId IdType="pubmed">27269431</ArticleId></ArticleIdList></Reference></ReferenceList></PubmedData></pubmed><affiliations><list><country><li>République populaire de Chine</li><li>États-Unis</li></country><region><li>Texas</li><li>État de New York</li></region></list><tree><country name="États-Unis"><region name="Texas"><name sortKey="Nyon, Mun Peak" sort="Nyon, Mun Peak" uniqKey="Nyon M" first="Mun Peak" last="Nyon">Mun Peak Nyon</name></region><name sortKey="Agrawal, Anurodh" sort="Agrawal, Anurodh" uniqKey="Agrawal A" first="Anurodh" last="Agrawal">Anurodh Agrawal</name><name sortKey="Algaissi, Abdullah" sort="Algaissi, Abdullah" uniqKey="Algaissi A" first="Abdullah" last="Algaissi">Abdullah Algaissi</name><name sortKey="Bottazzi, Maria Elena" sort="Bottazzi, Maria Elena" uniqKey="Bottazzi M" first="Maria Elena" last="Bottazzi">Maria Elena Bottazzi</name><name sortKey="Du, Lanying" sort="Du, Lanying" uniqKey="Du L" first="Lanying" last="Du">Lanying Du</name><name sortKey="Hotez, Peter J" sort="Hotez, Peter J" uniqKey="Hotez P" first="Peter J" last="Hotez">Peter J. Hotez</name><name sortKey="Naceanceno, Kevin S" sort="Naceanceno, Kevin S" uniqKey="Naceanceno K" first="Kevin S" last="Naceanceno">Kevin S. Naceanceno</name><name sortKey="Peng, Bi Hung" sort="Peng, Bi Hung" uniqKey="Peng B" first="Bi-Hung" last="Peng">Bi-Hung Peng</name><name sortKey="Pollet, Jeroen" sort="Pollet, Jeroen" uniqKey="Pollet J" first="Jeroen" last="Pollet">Jeroen Pollet</name><name sortKey="Seid, Christopher A" sort="Seid, Christopher A" uniqKey="Seid C" first="Christopher A" last="Seid">Christopher A. Seid</name><name sortKey="Strych, Ulrich" sort="Strych, Ulrich" uniqKey="Strych U" first="Ulrich" last="Strych">Ulrich Strych</name><name sortKey="Tai, Wanbo" sort="Tai, Wanbo" uniqKey="Tai W" first="Wanbo" last="Tai">Wanbo Tai</name><name sortKey="Tseng, Chien Te Kent" sort="Tseng, Chien Te Kent" uniqKey="Tseng C" first="Chien-Te Kent" last="Tseng">Chien-Te Kent Tseng</name></country><country name="République populaire de Chine"><noRegion><name sortKey="Jiang, Shibo" sort="Jiang, Shibo" uniqKey="Jiang S" first="Shibo" last="Jiang">Shibo Jiang</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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