Increased Antibody Affinity Confers Broad In Vitro Protection against Escape Mutants of Severe Acute Respiratory Syndrome Coronavirus
Identifieur interne : 002517 ( Ncbi/Merge ); précédent : 002516; suivant : 002518Increased Antibody Affinity Confers Broad In Vitro Protection against Escape Mutants of Severe Acute Respiratory Syndrome Coronavirus
Auteurs : Mridula Rani ; Meagan Bolles [États-Unis] ; Eric F. Donaldson [États-Unis] ; Thomas Van Blarcom ; Ralph Baric [États-Unis] ; Brent Iverson ; George Georgiou [États-Unis]Source :
- Journal of Virology [ 0022-538X ] ; 2012.
Descripteurs français
- KwdFr :
- Affinité des anticorps, Alignement de séquences, Animaux, Anticorps antiviraux (), Anticorps antiviraux (génétique), Anticorps antiviraux (immunologie), Anticorps antiviraux (pharmacologie), Anticorps neutralisants (), Anticorps neutralisants (génétique), Anticorps neutralisants (immunologie), Anticorps neutralisants (pharmacologie), Anticorps à chaîne unique (génétique), Anticorps à chaîne unique (immunologie), Anticorps à chaîne unique (pharmacologie), Cinétique, Données de séquences moléculaires, Humains, Lignée cellulaire, Mutation, Syndrome respiratoire aigu sévère (), Syndrome respiratoire aigu sévère (immunologie), Syndrome respiratoire aigu sévère (virologie), Séquence d'acides aminés, Tests de neutralisation, Virus du SRAS (), Virus du SRAS (génétique), Virus du SRAS (immunologie).
- MESH :
- génétique : Anticorps antiviraux, Anticorps neutralisants, Anticorps à chaîne unique, Virus du SRAS.
- immunologie : Anticorps antiviraux, Anticorps neutralisants, Anticorps à chaîne unique, Syndrome respiratoire aigu sévère, Virus du SRAS.
- pharmacologie : Anticorps antiviraux, Anticorps neutralisants, Anticorps à chaîne unique.
- virologie : Syndrome respiratoire aigu sévère.
- Affinité des anticorps, Alignement de séquences, Animaux, Anticorps antiviraux, Anticorps neutralisants, Cinétique, Données de séquences moléculaires, Humains, Lignée cellulaire, Mutation, Syndrome respiratoire aigu sévère, Séquence d'acides aminés, Tests de neutralisation, Virus du SRAS.
English descriptors
- KwdEn :
- Amino Acid Sequence, Animals, Antibodies, Neutralizing (chemistry), Antibodies, Neutralizing (genetics), Antibodies, Neutralizing (immunology), Antibodies, Neutralizing (pharmacology), Antibodies, Viral (chemistry), Antibodies, Viral (genetics), Antibodies, Viral (immunology), Antibodies, Viral (pharmacology), Antibody Affinity, Cell Line, Humans, Kinetics, Molecular Sequence Data, Mutation, Neutralization Tests, SARS Virus (chemistry), SARS Virus (drug effects), SARS Virus (genetics), SARS Virus (immunology), Sequence Alignment, Severe Acute Respiratory Syndrome (immunology), Severe Acute Respiratory Syndrome (prevention & control), Severe Acute Respiratory Syndrome (virology), Single-Chain Antibodies (genetics), Single-Chain Antibodies (immunology), Single-Chain Antibodies (pharmacology).
- MESH :
- chemical , chemistry : Antibodies, Neutralizing, Antibodies, Viral.
- chemical , genetics : Antibodies, Neutralizing, Antibodies, Viral, Single-Chain Antibodies.
- chemical , immunology : Antibodies, Neutralizing, Antibodies, Viral, Single-Chain Antibodies.
- chemical , pharmacology : Antibodies, Neutralizing, Antibodies, Viral, Single-Chain Antibodies.
- chemistry : SARS Virus.
- drug effects : SARS Virus.
- genetics : SARS Virus.
- immunology : SARS Virus, Severe Acute Respiratory Syndrome.
- prevention & control : Severe Acute Respiratory Syndrome.
- virology : Severe Acute Respiratory Syndrome.
- Amino Acid Sequence, Animals, Antibody Affinity, Cell Line, Humans, Kinetics, Molecular Sequence Data, Mutation, Neutralization Tests, Sequence Alignment.
Abstract
Even though the effect of antibody affinity on neutralization potency is well documented, surprisingly, its impact on neutralization breadth and escape has not been systematically determined. Here, random mutagenesis and DNA shuffling of the single-chain variable fragment of the neutralizing antibody 80R followed by bacterial display screening using anchored periplasmic expression (APEx) were used to generate a number of higher-affinity variants of the severe acute respiratory syndrome coronavirus (SARS-CoV)-neutralizing antibody 80R with equilibrium dissociation constants (
Url:
DOI: 10.1128/JVI.00233-12
PubMed: 22696652
PubMed Central: 3416138
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PMC:3416138Le document en format XML
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Protection against Escape Mutants of Severe Acute Respiratory Syndrome Coronavirus</title>
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<term>Animals</term>
<term>Antibodies, Neutralizing (chemistry)</term>
<term>Antibodies, Neutralizing (genetics)</term>
<term>Antibodies, Neutralizing (immunology)</term>
<term>Antibodies, Neutralizing (pharmacology)</term>
<term>Antibodies, Viral (chemistry)</term>
<term>Antibodies, Viral (genetics)</term>
<term>Antibodies, Viral (immunology)</term>
<term>Antibodies, Viral (pharmacology)</term>
<term>Antibody Affinity</term>
<term>Cell Line</term>
<term>Humans</term>
<term>Kinetics</term>
<term>Molecular Sequence Data</term>
<term>Mutation</term>
<term>Neutralization Tests</term>
<term>SARS Virus (chemistry)</term>
<term>SARS Virus (drug effects)</term>
<term>SARS Virus (genetics)</term>
<term>SARS Virus (immunology)</term>
<term>Sequence Alignment</term>
<term>Severe Acute Respiratory Syndrome (immunology)</term>
<term>Severe Acute Respiratory Syndrome (prevention & control)</term>
<term>Severe Acute Respiratory Syndrome (virology)</term>
<term>Single-Chain Antibodies (genetics)</term>
<term>Single-Chain Antibodies (immunology)</term>
<term>Single-Chain Antibodies (pharmacology)</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>Affinité des anticorps</term>
<term>Alignement de séquences</term>
<term>Animaux</term>
<term>Anticorps antiviraux ()</term>
<term>Anticorps antiviraux (génétique)</term>
<term>Anticorps antiviraux (immunologie)</term>
<term>Anticorps antiviraux (pharmacologie)</term>
<term>Anticorps neutralisants ()</term>
<term>Anticorps neutralisants (génétique)</term>
<term>Anticorps neutralisants (immunologie)</term>
<term>Anticorps neutralisants (pharmacologie)</term>
<term>Anticorps à chaîne unique (génétique)</term>
<term>Anticorps à chaîne unique (immunologie)</term>
<term>Anticorps à chaîne unique (pharmacologie)</term>
<term>Cinétique</term>
<term>Données de séquences moléculaires</term>
<term>Humains</term>
<term>Lignée cellulaire</term>
<term>Mutation</term>
<term>Syndrome respiratoire aigu sévère ()</term>
<term>Syndrome respiratoire aigu sévère (immunologie)</term>
<term>Syndrome respiratoire aigu sévère (virologie)</term>
<term>Séquence d'acides aminés</term>
<term>Tests de neutralisation</term>
<term>Virus du SRAS ()</term>
<term>Virus du SRAS (génétique)</term>
<term>Virus du SRAS (immunologie)</term>
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<keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Antibodies, Neutralizing</term>
<term>Antibodies, Viral</term>
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<term>Antibodies, Viral</term>
<term>Single-Chain Antibodies</term>
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<keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Antibodies, Neutralizing</term>
<term>Antibodies, Viral</term>
<term>Single-Chain Antibodies</term>
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<keywords scheme="MESH" type="chemical" qualifier="pharmacology" xml:lang="en"><term>Antibodies, Neutralizing</term>
<term>Antibodies, Viral</term>
<term>Single-Chain Antibodies</term>
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<keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>SARS Virus</term>
</keywords>
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</keywords>
<keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>SARS Virus</term>
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<term>Anticorps neutralisants</term>
<term>Anticorps à chaîne unique</term>
<term>Virus du SRAS</term>
</keywords>
<keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Anticorps antiviraux</term>
<term>Anticorps neutralisants</term>
<term>Anticorps à chaîne unique</term>
<term>Syndrome respiratoire aigu sévère</term>
<term>Virus du SRAS</term>
</keywords>
<keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>SARS Virus</term>
<term>Severe Acute Respiratory Syndrome</term>
</keywords>
<keywords scheme="MESH" qualifier="pharmacologie" xml:lang="fr"><term>Anticorps antiviraux</term>
<term>Anticorps neutralisants</term>
<term>Anticorps à chaîne unique</term>
</keywords>
<keywords scheme="MESH" qualifier="prevention & control" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term>
</keywords>
<keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Syndrome respiratoire aigu sévère</term>
</keywords>
<keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Amino Acid Sequence</term>
<term>Animals</term>
<term>Antibody Affinity</term>
<term>Cell Line</term>
<term>Humans</term>
<term>Kinetics</term>
<term>Molecular Sequence Data</term>
<term>Mutation</term>
<term>Neutralization Tests</term>
<term>Sequence Alignment</term>
</keywords>
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<term>Alignement de séquences</term>
<term>Animaux</term>
<term>Anticorps antiviraux</term>
<term>Anticorps neutralisants</term>
<term>Cinétique</term>
<term>Données de séquences moléculaires</term>
<term>Humains</term>
<term>Lignée cellulaire</term>
<term>Mutation</term>
<term>Syndrome respiratoire aigu sévère</term>
<term>Séquence d'acides aminés</term>
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<front><div type="abstract" xml:lang="en"><p>Even though the effect of antibody affinity on neutralization potency is well documented, surprisingly, its impact on neutralization breadth and escape has not been systematically determined. Here, random mutagenesis and DNA shuffling of the single-chain variable fragment of the neutralizing antibody 80R followed by bacterial display screening using anchored periplasmic expression (APEx) were used to generate a number of higher-affinity variants of the severe acute respiratory syndrome coronavirus (SARS-CoV)-neutralizing antibody 80R with equilibrium dissociation constants (<italic>K<sub>D</sub>
</italic>
) as low as 37 pM, a >270-fold improvement relative to that of the parental 80R single-chain variable fragment (scFv). As expected, antigen affinity was shown to correlate directly with neutralization potency toward the icUrbani strain of SARS-CoV. Additionally, the highest-affinity antibody fragment displayed 10-fold-increased broad neutralization <italic>in vitro</italic>
and completely protected against several SARS-CoV strains containing substitutions associated with antibody escape. Importantly, higher affinity also led to the suppression of viral escape mutants <italic>in vitro</italic>
. Escape from the highest-affinity variant required reduced selective pressure and multiple substitutions in the binding epitope. Collectively, these results support the hypothesis that engineered antibodies with picomolar dissociation constants for a neutralizing epitope can confer escape-resistant protection.</p>
</div>
</front>
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Protection against Escape Mutants of Severe Acute Respiratory Syndrome Coronavirus</title>
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<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Increased Antibody Affinity Confers Broad <italic>In Vitro</italic>
Protection against Escape Mutants of Severe Acute Respiratory Syndrome Coronavirus</title>
<author><name sortKey="Rani, Mridula" sort="Rani, Mridula" uniqKey="Rani M" first="Mridula" last="Rani">Mridula Rani</name>
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</affiliation>
</author>
<author><name sortKey="Bolles, Meagan" sort="Bolles, Meagan" uniqKey="Bolles M" first="Meagan" last="Bolles">Meagan Bolles</name>
<affiliation wicri:level="4"><nlm:aff id="aff6">Department of Microbiology and Immunology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Microbiology and Immunology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina</wicri:regionArea>
<placeName><region type="state">Caroline du Nord</region>
<settlement type="city">Chapel Hill (Caroline du Nord)</settlement>
</placeName>
<orgName type="university">Université de Caroline du Nord à Chapel Hill</orgName>
</affiliation>
</author>
<author><name sortKey="Donaldson, Eric F" sort="Donaldson, Eric F" uniqKey="Donaldson E" first="Eric F." last="Donaldson">Eric F. Donaldson</name>
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<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Epidemiology, School of Public Health, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina</wicri:regionArea>
<placeName><region type="state">Caroline du Nord</region>
<settlement type="city">Chapel Hill (Caroline du Nord)</settlement>
</placeName>
<orgName type="university">Université de Caroline du Nord à Chapel Hill</orgName>
</affiliation>
</author>
<author><name sortKey="Van Blarcom, Thomas" sort="Van Blarcom, Thomas" uniqKey="Van Blarcom T" first="Thomas" last="Van Blarcom">Thomas Van Blarcom</name>
<affiliation><nlm:aff id="aff2">Department of Chemical Engineering</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Baric, Ralph" sort="Baric, Ralph" uniqKey="Baric R" first="Ralph" last="Baric">Ralph Baric</name>
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<wicri:regionArea>Department of Epidemiology, School of Public Health, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina</wicri:regionArea>
<placeName><region type="state">Caroline du Nord</region>
<settlement type="city">Chapel Hill (Caroline du Nord)</settlement>
</placeName>
<orgName type="university">Université de Caroline du Nord à Chapel Hill</orgName>
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<author><name sortKey="Iverson, Brent" sort="Iverson, Brent" uniqKey="Iverson B" first="Brent" last="Iverson">Brent Iverson</name>
<affiliation><nlm:aff id="aff1">Institute of Cellular and Molecular Biology</nlm:aff>
</affiliation>
<affiliation><nlm:aff id="aff3">Department of Chemistry and Biochemistry</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Georgiou, George" sort="Georgiou, George" uniqKey="Georgiou G" first="George" last="Georgiou">George Georgiou</name>
<affiliation><nlm:aff id="aff1">Institute of Cellular and Molecular Biology</nlm:aff>
</affiliation>
<affiliation><nlm:aff id="aff2">Department of Chemical Engineering</nlm:aff>
</affiliation>
<affiliation wicri:level="4"><nlm:aff id="aff4">Department of Molecular Genetics and Microbiology, The University of Texas at Austin, Austin, Texas, USA</nlm:aff>
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<wicri:regionArea>Department of Molecular Genetics and Microbiology, The University of Texas at Austin, Austin, Texas</wicri:regionArea>
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<orgName type="university">Université du Texas à Austin</orgName>
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<series><title level="j">Journal of Virology</title>
<idno type="ISSN">0022-538X</idno>
<idno type="eISSN">1098-5514</idno>
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<front><div type="abstract" xml:lang="en"><p>Even though the effect of antibody affinity on neutralization potency is well documented, surprisingly, its impact on neutralization breadth and escape has not been systematically determined. Here, random mutagenesis and DNA shuffling of the single-chain variable fragment of the neutralizing antibody 80R followed by bacterial display screening using anchored periplasmic expression (APEx) were used to generate a number of higher-affinity variants of the severe acute respiratory syndrome coronavirus (SARS-CoV)-neutralizing antibody 80R with equilibrium dissociation constants (<italic>K<sub>D</sub>
</italic>
) as low as 37 pM, a >270-fold improvement relative to that of the parental 80R single-chain variable fragment (scFv). As expected, antigen affinity was shown to correlate directly with neutralization potency toward the icUrbani strain of SARS-CoV. Additionally, the highest-affinity antibody fragment displayed 10-fold-increased broad neutralization <italic>in vitro</italic>
and completely protected against several SARS-CoV strains containing substitutions associated with antibody escape. Importantly, higher affinity also led to the suppression of viral escape mutants <italic>in vitro</italic>
. Escape from the highest-affinity variant required reduced selective pressure and multiple substitutions in the binding epitope. Collectively, these results support the hypothesis that engineered antibodies with picomolar dissociation constants for a neutralizing epitope can confer escape-resistant protection.</p>
</div>
</front>
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<pubmed><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Increased antibody affinity confers broad in vitro protection against escape mutants of severe acute respiratory syndrome coronavirus.</title>
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<author><name sortKey="Bolles, Meagan" sort="Bolles, Meagan" uniqKey="Bolles M" first="Meagan" last="Bolles">Meagan Bolles</name>
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<term>Antibodies, Neutralizing (chemistry)</term>
<term>Antibodies, Neutralizing (genetics)</term>
<term>Antibodies, Neutralizing (immunology)</term>
<term>Antibodies, Neutralizing (pharmacology)</term>
<term>Antibodies, Viral (chemistry)</term>
<term>Antibodies, Viral (genetics)</term>
<term>Antibodies, Viral (immunology)</term>
<term>Antibodies, Viral (pharmacology)</term>
<term>Antibody Affinity</term>
<term>Cell Line</term>
<term>Humans</term>
<term>Kinetics</term>
<term>Molecular Sequence Data</term>
<term>Mutation</term>
<term>Neutralization Tests</term>
<term>SARS Virus (chemistry)</term>
<term>SARS Virus (drug effects)</term>
<term>SARS Virus (genetics)</term>
<term>SARS Virus (immunology)</term>
<term>Sequence Alignment</term>
<term>Severe Acute Respiratory Syndrome (immunology)</term>
<term>Severe Acute Respiratory Syndrome (prevention & control)</term>
<term>Severe Acute Respiratory Syndrome (virology)</term>
<term>Single-Chain Antibodies (genetics)</term>
<term>Single-Chain Antibodies (immunology)</term>
<term>Single-Chain Antibodies (pharmacology)</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>Affinité des anticorps</term>
<term>Alignement de séquences</term>
<term>Animaux</term>
<term>Anticorps antiviraux ()</term>
<term>Anticorps antiviraux (génétique)</term>
<term>Anticorps antiviraux (immunologie)</term>
<term>Anticorps antiviraux (pharmacologie)</term>
<term>Anticorps neutralisants ()</term>
<term>Anticorps neutralisants (génétique)</term>
<term>Anticorps neutralisants (immunologie)</term>
<term>Anticorps neutralisants (pharmacologie)</term>
<term>Anticorps à chaîne unique (génétique)</term>
<term>Anticorps à chaîne unique (immunologie)</term>
<term>Anticorps à chaîne unique (pharmacologie)</term>
<term>Cinétique</term>
<term>Données de séquences moléculaires</term>
<term>Humains</term>
<term>Lignée cellulaire</term>
<term>Mutation</term>
<term>Syndrome respiratoire aigu sévère ()</term>
<term>Syndrome respiratoire aigu sévère (immunologie)</term>
<term>Syndrome respiratoire aigu sévère (virologie)</term>
<term>Séquence d'acides aminés</term>
<term>Tests de neutralisation</term>
<term>Virus du SRAS ()</term>
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<term>Virus du SRAS (immunologie)</term>
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<term>Antibodies, Viral</term>
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<term>Antibodies, Viral</term>
<term>Single-Chain Antibodies</term>
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<term>Antibodies, Viral</term>
<term>Single-Chain Antibodies</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="pharmacology" xml:lang="en"><term>Antibodies, Neutralizing</term>
<term>Antibodies, Viral</term>
<term>Single-Chain Antibodies</term>
</keywords>
<keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>SARS Virus</term>
</keywords>
<keywords scheme="MESH" qualifier="drug effects" xml:lang="en"><term>SARS Virus</term>
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<term>Anticorps neutralisants</term>
<term>Anticorps à chaîne unique</term>
<term>Virus du SRAS</term>
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<term>Anticorps neutralisants</term>
<term>Anticorps à chaîne unique</term>
<term>Syndrome respiratoire aigu sévère</term>
<term>Virus du SRAS</term>
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<term>Severe Acute Respiratory Syndrome</term>
</keywords>
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<term>Anticorps neutralisants</term>
<term>Anticorps à chaîne unique</term>
</keywords>
<keywords scheme="MESH" qualifier="prevention & control" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term>
</keywords>
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</keywords>
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<term>Animals</term>
<term>Antibody Affinity</term>
<term>Cell Line</term>
<term>Humans</term>
<term>Kinetics</term>
<term>Molecular Sequence Data</term>
<term>Mutation</term>
<term>Neutralization Tests</term>
<term>Sequence Alignment</term>
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<term>Alignement de séquences</term>
<term>Animaux</term>
<term>Anticorps antiviraux</term>
<term>Anticorps neutralisants</term>
<term>Cinétique</term>
<term>Données de séquences moléculaires</term>
<term>Humains</term>
<term>Lignée cellulaire</term>
<term>Mutation</term>
<term>Syndrome respiratoire aigu sévère</term>
<term>Séquence d'acides aminés</term>
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<front><div type="abstract" xml:lang="en">Even though the effect of antibody affinity on neutralization potency is well documented, surprisingly, its impact on neutralization breadth and escape has not been systematically determined. Here, random mutagenesis and DNA shuffling of the single-chain variable fragment of the neutralizing antibody 80R followed by bacterial display screening using anchored periplasmic expression (APEx) were used to generate a number of higher-affinity variants of the severe acute respiratory syndrome coronavirus (SARS-CoV)-neutralizing antibody 80R with equilibrium dissociation constants (K(D)) as low as 37 pM, a >270-fold improvement relative to that of the parental 80R single-chain variable fragment (scFv). As expected, antigen affinity was shown to correlate directly with neutralization potency toward the icUrbani strain of SARS-CoV. Additionally, the highest-affinity antibody fragment displayed 10-fold-increased broad neutralization in vitro and completely protected against several SARS-CoV strains containing substitutions associated with antibody escape. Importantly, higher affinity also led to the suppression of viral escape mutants in vitro. Escape from the highest-affinity variant required reduced selective pressure and multiple substitutions in the binding epitope. Collectively, these results support the hypothesis that engineered antibodies with picomolar dissociation constants for a neutralizing epitope can confer escape-resistant protection.</div>
</front>
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