Rapid discovery and optimization of therapeutic antibodies against emerging infectious diseases.
Identifieur interne : 001C60 ( Ncbi/Curation ); précédent : 001C59; suivant : 001C61Rapid discovery and optimization of therapeutic antibodies against emerging infectious diseases.
Auteurs : J. Rogers [États-Unis] ; R J Schoepp ; O. Schröder ; T L Clements ; T F Holland ; J Q Li ; J. Li ; L M Lewis ; R P Dirmeier ; G J Frey ; X. Tan ; K. Wong ; G. Woodnutt ; M. Keller ; D S Reed ; B E Kimmel ; E C TozerSource :
- Protein engineering, design & selection : PEDS [ 1741-0126 ] ; 2008.
Descripteurs français
- KwdFr :
- Animaux, Anticorps antiviraux (génétique), Anticorps antiviraux (isolement et purification), Anticorps antiviraux (usage thérapeutique), Cellules Vero, Découverte de médicament, Humains, Maladies transmissibles émergentes (), Mutation ponctuelle (immunologie), Souris, Souris de lignée BALB C, Spécificité des anticorps (immunologie), Syndrome respiratoire aigu sévère (), Tests de neutralisation, Virus du SRAS (immunologie), Évolution moléculaire dirigée ().
- MESH :
- génétique : Anticorps antiviraux.
- immunologie : Mutation ponctuelle, Spécificité des anticorps, Virus du SRAS.
- isolement et purification : Anticorps antiviraux.
- usage thérapeutique : Anticorps antiviraux.
- Animaux, Cellules Vero, Découverte de médicament, Humains, Maladies transmissibles émergentes, Souris, Souris de lignée BALB C, Syndrome respiratoire aigu sévère, Tests de neutralisation, Évolution moléculaire dirigée.
English descriptors
- KwdEn :
- Animals, Antibodies, Viral (genetics), Antibodies, Viral (isolation & purification), Antibodies, Viral (therapeutic use), Antibody Specificity (immunology), Chlorocebus aethiops, Communicable Diseases, Emerging (prevention & control), Communicable Diseases, Emerging (therapy), Directed Molecular Evolution (methods), Drug Discovery, Humans, Mice, Mice, Inbred BALB C, Neutralization Tests, Point Mutation (immunology), SARS Virus (immunology), Severe Acute Respiratory Syndrome (prevention & control), Severe Acute Respiratory Syndrome (therapy), Vero Cells.
- MESH :
- chemical , genetics : Antibodies, Viral.
- chemical , isolation & purification : Antibodies, Viral.
- chemical , therapeutic use : Antibodies, Viral.
- immunology : Antibody Specificity, Point Mutation, SARS Virus.
- methods : Directed Molecular Evolution.
- prevention & control : Communicable Diseases, Emerging, Severe Acute Respiratory Syndrome.
- therapy : Communicable Diseases, Emerging, Severe Acute Respiratory Syndrome.
- Animals, Chlorocebus aethiops, Drug Discovery, Humans, Mice, Mice, Inbred BALB C, Neutralization Tests, Vero Cells.
Abstract
Using a comprehensive set of discovery and optimization tools, antibodies were produced with the ability to neutralize SARS coronavirus (SARS-CoV) infection in Vero E6 cells and in animal models. These anti-SARS antibodies were discovered using a novel DNA display method, which can identify new antibodies within days. Once neutralizing antibodies were identified, a comprehensive and effective means of converting the mouse sequences to human frameworks was accomplished using HuFR (human framework reassembly) technology. The best variant (61G4) from this screen showed a 3.5-4-fold improvement in neutralization of SARS-CoV infection in vitro. Finally, using a complete site-saturation mutagenesis methodology focused on the CDR (complementarity determining regions), a single point mutation (51E7) was identified that improved the 80% plaque reduction neutralization of the virus by greater than 8-fold. These discovery and evolution strategies can be applied to any emerging pathogen or toxin where a causative agent is known.
DOI: 10.1093/protein/gzn027
PubMed: 18480090
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pubmed:18480090Le document en format XML
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<term>Antibodies, Viral (isolation & purification)</term>
<term>Antibodies, Viral (therapeutic use)</term>
<term>Antibody Specificity (immunology)</term>
<term>Chlorocebus aethiops</term>
<term>Communicable Diseases, Emerging (prevention & control)</term>
<term>Communicable Diseases, Emerging (therapy)</term>
<term>Directed Molecular Evolution (methods)</term>
<term>Drug Discovery</term>
<term>Humans</term>
<term>Mice</term>
<term>Mice, Inbred BALB C</term>
<term>Neutralization Tests</term>
<term>Point Mutation (immunology)</term>
<term>SARS Virus (immunology)</term>
<term>Severe Acute Respiratory Syndrome (prevention & control)</term>
<term>Severe Acute Respiratory Syndrome (therapy)</term>
<term>Vero Cells</term>
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<term>Anticorps antiviraux (génétique)</term>
<term>Anticorps antiviraux (isolement et purification)</term>
<term>Anticorps antiviraux (usage thérapeutique)</term>
<term>Cellules Vero</term>
<term>Découverte de médicament</term>
<term>Humains</term>
<term>Maladies transmissibles émergentes ()</term>
<term>Mutation ponctuelle (immunologie)</term>
<term>Souris</term>
<term>Souris de lignée BALB C</term>
<term>Spécificité des anticorps (immunologie)</term>
<term>Syndrome respiratoire aigu sévère ()</term>
<term>Tests de neutralisation</term>
<term>Virus du SRAS (immunologie)</term>
<term>Évolution moléculaire dirigée ()</term>
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<keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>Antibodies, Viral</term>
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<keywords scheme="MESH" type="chemical" qualifier="therapeutic use" xml:lang="en"><term>Antibodies, Viral</term>
</keywords>
<keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Anticorps antiviraux</term>
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<keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Mutation ponctuelle</term>
<term>Spécificité des anticorps</term>
<term>Virus du SRAS</term>
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<keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>Antibody Specificity</term>
<term>Point Mutation</term>
<term>SARS Virus</term>
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<keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Anticorps antiviraux</term>
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<term>Severe Acute Respiratory Syndrome</term>
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<keywords scheme="MESH" qualifier="therapy" xml:lang="en"><term>Communicable Diseases, Emerging</term>
<term>Severe Acute Respiratory Syndrome</term>
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</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Animals</term>
<term>Chlorocebus aethiops</term>
<term>Drug Discovery</term>
<term>Humans</term>
<term>Mice</term>
<term>Mice, Inbred BALB C</term>
<term>Neutralization Tests</term>
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<term>Maladies transmissibles émergentes</term>
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<term>Souris de lignée BALB C</term>
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<front><div type="abstract" xml:lang="en">Using a comprehensive set of discovery and optimization tools, antibodies were produced with the ability to neutralize SARS coronavirus (SARS-CoV) infection in Vero E6 cells and in animal models. These anti-SARS antibodies were discovered using a novel DNA display method, which can identify new antibodies within days. Once neutralizing antibodies were identified, a comprehensive and effective means of converting the mouse sequences to human frameworks was accomplished using HuFR (human framework reassembly) technology. The best variant (61G4) from this screen showed a 3.5-4-fold improvement in neutralization of SARS-CoV infection in vitro. Finally, using a complete site-saturation mutagenesis methodology focused on the CDR (complementarity determining regions), a single point mutation (51E7) was identified that improved the 80% plaque reduction neutralization of the virus by greater than 8-fold. These discovery and evolution strategies can be applied to any emerging pathogen or toxin where a causative agent is known.</div>
</front>
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