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Article: Development of a safe and convenient neutralization assay for rapid screening of influenza HA-specific neutralizing monoclonal antibodies

TitleDevelopment of a safe and convenient neutralization assay for rapid screening of influenza HA-specific neutralizing monoclonal antibodies
Authors
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
Citation
Biochemical And Biophysical Research Communications, 2010, v. 397 n. 3, p. 580-585 How to Cite?
Abstract
The 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.
Persistent Identifierhttp://hdl.handle.net/10722/135276
ISSN
2013 Impact Factor: 2.281
ISI Accession Number ID
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).

References

 

DC FieldValueLanguage
dc.contributor.authorDu, Len_HK
dc.contributor.authorZhao, Gen_HK
dc.contributor.authorZhang, Xen_HK
dc.contributor.authorLiu, Zen_HK
dc.contributor.authorYu, Hen_HK
dc.contributor.authorZheng, BJen_HK
dc.contributor.authorZhou, Yen_HK
dc.contributor.authorJiang, Sen_HK
dc.date.accessioned2011-07-27T01:31:04Z-
dc.date.available2011-07-27T01:31:04Z-
dc.date.issued2010en_HK
dc.identifier.citationBiochemical And Biophysical Research Communications, 2010, v. 397 n. 3, p. 580-585en_HK
dc.identifier.issn0006-291Xen_HK
dc.identifier.urihttp://hdl.handle.net/10722/135276-
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.en_HK
dc.languageengen_US
dc.publisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/wps/find/journaldescription.cws_home/622790/descriptionen_HK
dc.relation.ispartofBiochemical and Biophysical Research Communicationsen_HK
dc.subjectHemagglutininen_HK
dc.subjectInfluenza A virusen_HK
dc.subjectMonoclonal antibodiesen_HK
dc.subjectPseudovirus neutralization assayen_HK
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.titleDevelopment of a safe and convenient neutralization assay for rapid screening of influenza HA-specific neutralizing monoclonal antibodiesen_HK
dc.typeArticleen_HK
dc.identifier.emailZheng, BJ:bzheng@hkucc.hku.hken_HK
dc.identifier.authorityZheng, BJ=rp00353en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1016/j.bbrc.2010.05.161en_HK
dc.identifier.pmid20617558en_HK
dc.identifier.scopuseid_2-s2.0-77954213672en_HK
dc.identifier.hkuros188634en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-77954213672&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume397en_HK
dc.identifier.issue3en_HK
dc.identifier.spage580en_HK
dc.identifier.epage585en_HK
dc.identifier.isiWOS:000279974600038-
dc.publisher.placeUnited Statesen_HK
dc.identifier.scopusauthoridDu, L=8686996200en_HK
dc.identifier.scopusauthoridZhao, G=8684553000en_HK
dc.identifier.scopusauthoridZhang, X=23112057700en_HK
dc.identifier.scopusauthoridLiu, Z=16319107300en_HK
dc.identifier.scopusauthoridYu, H=36349449900en_HK
dc.identifier.scopusauthoridZheng, BJ=7201780588en_HK
dc.identifier.scopusauthoridZhou, Y=8791655300en_HK
dc.identifier.scopusauthoridJiang, S=7404453146en_HK
dc.identifier.citeulike7272693-

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