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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
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TitleImproved breadth and potency of an HIV-1-neutralizing human single-chain antibody by random mutagenesis and sequential antigen panning
 
AuthorsZhang, MY1 3
Shu, Y1
Rudolph, D4
Prabakaran, P1
Labrijn, AF2
Zwick, MB2
Lal, RB4
Dimitrov, DS1
 
Issue Date2004
 
PublisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/locate/jmb
 
CitationJournal Of Molecular Biology, 2004, v. 335 n. 1, p. 209-219 [How to Cite?]
DOI: http://dx.doi.org/10.1016/j.jmb.2003.09.055
 
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.
 
ISSN0022-2836
2012 Impact Factor: 3.905
2012 SCImago Journal Rankings: 2.509
 
DOIhttp://dx.doi.org/10.1016/j.jmb.2003.09.055
 
ISI Accession Number IDWOS:000187879600016
 
ReferencesReferences in Scopus
 
DC FieldValue
dc.contributor.authorZhang, MY
 
dc.contributor.authorShu, Y
 
dc.contributor.authorRudolph, D
 
dc.contributor.authorPrabakaran, P
 
dc.contributor.authorLabrijn, AF
 
dc.contributor.authorZwick, MB
 
dc.contributor.authorLal, RB
 
dc.contributor.authorDimitrov, DS
 
dc.date.accessioned2012-08-08T08:49:26Z
 
dc.date.available2012-08-08T08:49:26Z
 
dc.date.issued2004
 
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.
 
dc.description.natureLink_to_subscribed_fulltext
 
dc.identifier.citationJournal Of Molecular Biology, 2004, v. 335 n. 1, p. 209-219 [How to Cite?]
DOI: http://dx.doi.org/10.1016/j.jmb.2003.09.055
 
dc.identifier.doihttp://dx.doi.org/10.1016/j.jmb.2003.09.055
 
dc.identifier.epage219
 
dc.identifier.isiWOS:000187879600016
 
dc.identifier.issn0022-2836
2012 Impact Factor: 3.905
2012 SCImago Journal Rankings: 2.509
 
dc.identifier.issue1
 
dc.identifier.pmid14659751
 
dc.identifier.scopuseid_2-s2.0-0344255655
 
dc.identifier.spage209
 
dc.identifier.urihttp://hdl.handle.net/10722/157375
 
dc.identifier.volume335
 
dc.languageeng
 
dc.publisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/locate/jmb
 
dc.publisher.placeUnited Kingdom
 
dc.relation.ispartofJournal of Molecular Biology
 
dc.relation.referencesReferences in Scopus
 
dc.subject.meshAntibody Affinity
 
dc.subject.meshAntibody Specificity
 
dc.subject.meshAntigens, Cd4 - Immunology
 
dc.subject.meshDrug Evaluation, Preclinical - Methods
 
dc.subject.meshHiv Antibodies - Genetics - Immunology
 
dc.subject.meshHiv Antigens - Immunology
 
dc.subject.meshHiv Envelope Protein Gp120 - Immunology
 
dc.subject.meshHiv-1 - Immunology
 
dc.subject.meshHumans
 
dc.subject.meshImmunoglobulin Fragments - Genetics - Immunology
 
dc.subject.meshInhibitory Concentration 50
 
dc.subject.meshMutagenesis
 
dc.subject.meshMutation
 
dc.subject.meshPeptide Library
 
dc.titleImproved breadth and potency of an HIV-1-neutralizing human single-chain antibody by random mutagenesis and sequential antigen panning
 
dc.typeArticle
 
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Author Affiliations
  1. National Cancer Institute at Frederick
  2. Scripps Research Institute
  3. SAIC-Frederick
  4. Coordinating Center for Infectious Diseases