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Article: Development of a safe and convenient neutralization assay for rapid screening of influenza HA-specific neutralizing monoclonal antibodies
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TitleDevelopment of a safe and convenient neutralization assay for rapid screening of influenza HA-specific neutralizing monoclonal antibodies
 
AuthorsDu, L3
Zhao, G1
Zhang, X3
Liu, Z3
Yu, H1
Zheng, BJ2
Zhou, Y1
Jiang, S3
 
KeywordsHemagglutinin
Influenza A virus
Monoclonal antibodies
Pseudovirus neutralization assay
 
Issue Date2010
 
PublisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/wps/find/journaldescription.cws_home/622790/description
 
CitationBiochemical And Biophysical Research Communications, 2010, v. 397 n. 3, p. 580-585 [How to Cite?]
DOI: http://dx.doi.org/10.1016/j.bbrc.2010.05.161
 
AbstractThe worldwide outbreak of the swine-origin 2009 H1N1 influenza A virus (IAV) and an increasing number of influenza cases caused by a highly pathogenic avian influenza (HPAI) H5N1 have accelerated the need to develop vaccines and antiviral agents against IAVs. Among various antivirals, neutralizing monoclonal antibodies (mAbs) are considered important passive therapeutics having an immediate effect against viral pathogens. Here we report a pseudovirus neutralization assay for rapid screening of neutralizing mAbs targeting hemagglutinin (HA) of H5N1 and H1N1 IAV. In this study, we generated six pseudoviruses with an HIV-1 backbone, respectively, expressing HA of four clades of H5N1 IAV and the 2009 epidemic H1N1 IAV. The resulting pseudoviruses were able to infect a variety of human and non-human cells, with 293T cells from human kidney as the most susceptible target cells. Using the established pseudovirus neutralization assay, we showed that three of ten selected mAbs specific to HA could potently neutralize infection of a pseudovirus bearing HA from the homologous IAV A/VietNam/1194/2004(H5N1) strain. This was highly consistent with the result of a microneutralization assay testing the same strain of a live IAV. Since the pseudovirus neutralization assay does not involve an infectious virus and can be performed without the requirement of a biosafety-3 laboratory, it may be applied for safe and rapid screening of neutralizing mAbs and antiviral agents targeting HA of IAVs. © 2010 Elsevier Inc.
 
ISSN0006-291X
2012 Impact Factor: 2.406
2012 SCImago Journal Rankings: 1.049
 
DOIhttp://dx.doi.org/10.1016/j.bbrc.2010.05.161
 
ISI Accession Number IDWOS:000279974600038
Funding AgencyGrant Number
National High Technology R&D Program of China2006AA02Z406
National Basic Research Program of China2005CB523001
National Natural Science Foundation of China30901371
Science Research2009ZX10004-4001
Funding Information:

This study was supported by the National High Technology R&D Program of China (863 Program, No. 2006AA02Z406), National Basic Research Program of China (973 Program, No. 2005CB523001), National Natural Science Foundation of China (30901371), and Mega-projects of Science Research for the 11th Five-Year Plan (2009ZX10004-4001).

 
ReferencesReferences in Scopus
 
DC FieldValue
dc.contributor.authorDu, L
 
dc.contributor.authorZhao, G
 
dc.contributor.authorZhang, X
 
dc.contributor.authorLiu, Z
 
dc.contributor.authorYu, H
 
dc.contributor.authorZheng, BJ
 
dc.contributor.authorZhou, Y
 
dc.contributor.authorJiang, S
 
dc.date.accessioned2011-07-27T01:31:04Z
 
dc.date.available2011-07-27T01:31:04Z
 
dc.date.issued2010
 
dc.description.abstractThe worldwide outbreak of the swine-origin 2009 H1N1 influenza A virus (IAV) and an increasing number of influenza cases caused by a highly pathogenic avian influenza (HPAI) H5N1 have accelerated the need to develop vaccines and antiviral agents against IAVs. Among various antivirals, neutralizing monoclonal antibodies (mAbs) are considered important passive therapeutics having an immediate effect against viral pathogens. Here we report a pseudovirus neutralization assay for rapid screening of neutralizing mAbs targeting hemagglutinin (HA) of H5N1 and H1N1 IAV. In this study, we generated six pseudoviruses with an HIV-1 backbone, respectively, expressing HA of four clades of H5N1 IAV and the 2009 epidemic H1N1 IAV. The resulting pseudoviruses were able to infect a variety of human and non-human cells, with 293T cells from human kidney as the most susceptible target cells. Using the established pseudovirus neutralization assay, we showed that three of ten selected mAbs specific to HA could potently neutralize infection of a pseudovirus bearing HA from the homologous IAV A/VietNam/1194/2004(H5N1) strain. This was highly consistent with the result of a microneutralization assay testing the same strain of a live IAV. Since the pseudovirus neutralization assay does not involve an infectious virus and can be performed without the requirement of a biosafety-3 laboratory, it may be applied for safe and rapid screening of neutralizing mAbs and antiviral agents targeting HA of IAVs. © 2010 Elsevier Inc.
 
dc.description.natureLink_to_subscribed_fulltext
 
dc.identifier.citationBiochemical And Biophysical Research Communications, 2010, v. 397 n. 3, p. 580-585 [How to Cite?]
DOI: http://dx.doi.org/10.1016/j.bbrc.2010.05.161
 
dc.identifier.citeulike7272693
 
dc.identifier.doihttp://dx.doi.org/10.1016/j.bbrc.2010.05.161
 
dc.identifier.epage585
 
dc.identifier.hkuros188634
 
dc.identifier.isiWOS:000279974600038
Funding AgencyGrant Number
National High Technology R&D Program of China2006AA02Z406
National Basic Research Program of China2005CB523001
National Natural Science Foundation of China30901371
Science Research2009ZX10004-4001
Funding Information:

This study was supported by the National High Technology R&D Program of China (863 Program, No. 2006AA02Z406), National Basic Research Program of China (973 Program, No. 2005CB523001), National Natural Science Foundation of China (30901371), and Mega-projects of Science Research for the 11th Five-Year Plan (2009ZX10004-4001).

 
dc.identifier.issn0006-291X
2012 Impact Factor: 2.406
2012 SCImago Journal Rankings: 1.049
 
dc.identifier.issue3
 
dc.identifier.pmid20617558
 
dc.identifier.scopuseid_2-s2.0-77954213672
 
dc.identifier.spage580
 
dc.identifier.urihttp://hdl.handle.net/10722/135276
 
dc.identifier.volume397
 
dc.languageeng
 
dc.publisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/wps/find/journaldescription.cws_home/622790/description
 
dc.publisher.placeUnited States
 
dc.relation.ispartofBiochemical and Biophysical Research Communications
 
dc.relation.referencesReferences in Scopus
 
dc.subject.meshAntibodies, Monoclonal - isolation and purification - therapeutic use
 
dc.subject.meshAntibodies, Neutralizing - isolation and purification - therapeutic use
 
dc.subject.meshHemagglutinin Glycoproteins, Influenza Virus - immunology
 
dc.subject.meshInfluenza, Human - immunology - therapy
 
dc.subject.meshNeutralization Tests
 
dc.subjectHemagglutinin
 
dc.subjectInfluenza A virus
 
dc.subjectMonoclonal antibodies
 
dc.subjectPseudovirus neutralization assay
 
dc.titleDevelopment of a safe and convenient neutralization assay for rapid screening of influenza HA-specific neutralizing monoclonal antibodies
 
dc.typeArticle
 
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Author Affiliations
  1. Institute of Microbiology Chinese Academy of Sciences
  2. The University of Hong Kong
  3. New York Blood Center