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Article: Improved breadth and potency of an HIV-1-neutralizing human single-chain antibody by random mutagenesis and sequential antigen panning

TitleImproved breadth and potency of an HIV-1-neutralizing human single-chain antibody by random mutagenesis and sequential antigen panning
Authors
Issue Date2004
PublisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/locate/jmb
Citation
Journal Of Molecular Biology, 2004, v. 335 n. 1, p. 209-219 How to Cite?
AbstractSeveral human monoclonal antibodies can neutralize a range of human immunodeficiency virus type 1 (HIV-1) primary isolates but their potency and related ability to suppress generation of HIV-1 escape mutants is significantly lower than the activity of antiretroviral drugs currently in clinical use. Recently, a human Fab, X5, was identified and found to neutralize primary isolates from different clades. Further improvement of the potency and breadth of HIV-1 neutralization by this antibody could be critical for its potential use in the treatment of HIV-1-infected patients. However, increasing potency of an antibody by selection from libraries may lead to a decrease in the breadth of neutralization. In an attempt to solve this problem, we subjected a random mutagenesis library of the scFv X5 to sequential rounds of selection on non-homologous HIV-1 envelope glycoproteins (Envs) dubbed sequential antigen panning (SAP). By using SAP, we identified two scFv antibodies, m6 and m9, that were tested with a panel of 33 diverse primary HIV-1 infectious isolates in an assay based on a reporter cell-line expressing high levels of CD4, CCR5 and CXCR4. The IC50 was less than 50 μg/ml for 21 (m6) and 19 (m9) out of 29 isolates from group M (subtypes A-C, F, G and CRF-01AE) and one isolate from group N; three isolates from group O were not significantly inhibited at 50 μg/ml. The average IC50 values for the two antibodies were significantly (p<0.001, n=29) lower compared to scFv X5. Their inhibitory activity does not appear to be related to the HIV-1 subtype, coreceptor usage or the disease stage. m9 inhibited infection of peripheral blood mononuclear cells by the primary isolates JRCSF, 89.6 and BR020 with IC90 of 4, 6 and 25 μg/ml, respectively; for a single-round infection by pseudovirus, the IC90 for JRSCF, 89.6, YU2 and HXBc2 was 15, 5, 15 and 5 μg/ml, respectively. In these two assays the IC 90 for m9 was, on average, two- to threefold lower than for scFv X5. These results demonstrate that both the potency and the breadth of HIV-1 neutralization of one of the few known potent broadly cross-reactive human monoclonal antibodies, scFv X5, could be improved significantly. However, only experiments in animal models and clinical trials in humans will show whether these new scFvs and the approach for their identification have potential in the development of prophylactics and therapeutics for HIV-1 infections. © 2003 Elsevier Ltd. All rights reserved.
Persistent Identifierhttp://hdl.handle.net/10722/157375
ISSN
2014 Impact Factor: 4.333
2013 SCImago Journal Rankings: 3.158
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorZhang, MYen_US
dc.contributor.authorShu, Yen_US
dc.contributor.authorRudolph, Den_US
dc.contributor.authorPrabakaran, Pen_US
dc.contributor.authorLabrijn, AFen_US
dc.contributor.authorZwick, MBen_US
dc.contributor.authorLal, RBen_US
dc.contributor.authorDimitrov, DSen_US
dc.date.accessioned2012-08-08T08:49:26Z-
dc.date.available2012-08-08T08:49:26Z-
dc.date.issued2004en_US
dc.identifier.citationJournal Of Molecular Biology, 2004, v. 335 n. 1, p. 209-219en_US
dc.identifier.issn0022-2836en_US
dc.identifier.urihttp://hdl.handle.net/10722/157375-
dc.description.abstractSeveral human monoclonal antibodies can neutralize a range of human immunodeficiency virus type 1 (HIV-1) primary isolates but their potency and related ability to suppress generation of HIV-1 escape mutants is significantly lower than the activity of antiretroviral drugs currently in clinical use. Recently, a human Fab, X5, was identified and found to neutralize primary isolates from different clades. Further improvement of the potency and breadth of HIV-1 neutralization by this antibody could be critical for its potential use in the treatment of HIV-1-infected patients. However, increasing potency of an antibody by selection from libraries may lead to a decrease in the breadth of neutralization. In an attempt to solve this problem, we subjected a random mutagenesis library of the scFv X5 to sequential rounds of selection on non-homologous HIV-1 envelope glycoproteins (Envs) dubbed sequential antigen panning (SAP). By using SAP, we identified two scFv antibodies, m6 and m9, that were tested with a panel of 33 diverse primary HIV-1 infectious isolates in an assay based on a reporter cell-line expressing high levels of CD4, CCR5 and CXCR4. The IC50 was less than 50 μg/ml for 21 (m6) and 19 (m9) out of 29 isolates from group M (subtypes A-C, F, G and CRF-01AE) and one isolate from group N; three isolates from group O were not significantly inhibited at 50 μg/ml. The average IC50 values for the two antibodies were significantly (p<0.001, n=29) lower compared to scFv X5. Their inhibitory activity does not appear to be related to the HIV-1 subtype, coreceptor usage or the disease stage. m9 inhibited infection of peripheral blood mononuclear cells by the primary isolates JRCSF, 89.6 and BR020 with IC90 of 4, 6 and 25 μg/ml, respectively; for a single-round infection by pseudovirus, the IC90 for JRSCF, 89.6, YU2 and HXBc2 was 15, 5, 15 and 5 μg/ml, respectively. In these two assays the IC 90 for m9 was, on average, two- to threefold lower than for scFv X5. These results demonstrate that both the potency and the breadth of HIV-1 neutralization of one of the few known potent broadly cross-reactive human monoclonal antibodies, scFv X5, could be improved significantly. However, only experiments in animal models and clinical trials in humans will show whether these new scFvs and the approach for their identification have potential in the development of prophylactics and therapeutics for HIV-1 infections. © 2003 Elsevier Ltd. All rights reserved.en_US
dc.languageengen_US
dc.publisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/locate/jmben_US
dc.relation.ispartofJournal of Molecular Biologyen_US
dc.subject.meshAntibody Affinityen_US
dc.subject.meshAntibody Specificityen_US
dc.subject.meshAntigens, Cd4 - Immunologyen_US
dc.subject.meshDrug Evaluation, Preclinical - Methodsen_US
dc.subject.meshHiv Antibodies - Genetics - Immunologyen_US
dc.subject.meshHiv Antigens - Immunologyen_US
dc.subject.meshHiv Envelope Protein Gp120 - Immunologyen_US
dc.subject.meshHiv-1 - Immunologyen_US
dc.subject.meshHumansen_US
dc.subject.meshImmunoglobulin Fragments - Genetics - Immunologyen_US
dc.subject.meshInhibitory Concentration 50en_US
dc.subject.meshMutagenesisen_US
dc.subject.meshMutationen_US
dc.subject.meshPeptide Libraryen_US
dc.titleImproved breadth and potency of an HIV-1-neutralizing human single-chain antibody by random mutagenesis and sequential antigen panningen_US
dc.typeArticleen_US
dc.identifier.emailZhang, MY:zhangmy@hku.hken_US
dc.identifier.authorityZhang, MY=rp01409en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1016/j.jmb.2003.09.055en_US
dc.identifier.pmid14659751en_US
dc.identifier.scopuseid_2-s2.0-0344255655en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0344255655&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume335en_US
dc.identifier.issue1en_US
dc.identifier.spage209en_US
dc.identifier.epage219en_US
dc.identifier.isiWOS:000187879600016-
dc.publisher.placeUnited Kingdomen_US
dc.identifier.scopusauthoridZhang, MY=35316639300en_US
dc.identifier.scopusauthoridShu, Y=7103239429en_US
dc.identifier.scopusauthoridRudolph, D=7101736051en_US
dc.identifier.scopusauthoridPrabakaran, P=6603634046en_US
dc.identifier.scopusauthoridLabrijn, AF=6507797385en_US
dc.identifier.scopusauthoridZwick, MB=8105899100en_US
dc.identifier.scopusauthoridLal, RB=35495076000en_US
dc.identifier.scopusauthoridDimitrov, DS=7202564539en_US

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