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Article: GST fusion proteins cause false positives during selection of viral movement protein specific single chain antibodies

TitleGST fusion proteins cause false positives during selection of viral movement protein specific single chain antibodies
Authors
Issue Date2001
PublisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/jviromet
Citation
Journal Of Virological Methods, 2001, v. 91 n. 2, p. 139-147 How to Cite?
AbstractGlutathione S-transferase (GST) fusion proteins are used frequently for investigating protein-protein and protein-DNA interactions. The present study demonstrates that the use of GST fusion proteins caused false positives during selection of phage-displayed single-chain antibody fragments (scFvs) specific for three domains of the movement protein (NSM) of tomato spotted wilt virus (TSWV). To identify and exclude the false positives when using GST as a fusion partner linked to the antigen of interest, indirect phage enzyme-linked immunosorbent assay (ELISA) was compared with capture phage ELISA. Of 210 enriched phage clones, indirect phage ELISA identified 106 clones specific for binding to GST-domain fusions but not to GST. In contrast, using capture phage ELISA, all 106 selected clones were identified as false positives, reacting with the GST fusion proteins and GST. This was confirmed by characterization of soluble scFv antibodies. The data indicate that GST fusion proteins seem unsuitable for screening of phage-displayed antibody fragments and it is essential to use capture phage ELISA, instead of the indirect phage ELISA used commonly to exclude false positives in characterization of selected clones with GST fusion proteins. © 2001 Elsevier Science B.V.
Persistent Identifierhttp://hdl.handle.net/10722/157331
ISSN
2015 Impact Factor: 1.508
2015 SCImago Journal Rankings: 0.866
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorZhang, MYen_US
dc.contributor.authorSchillberg, Sen_US
dc.contributor.authorZimmermann, Sen_US
dc.contributor.authorLiao, YCen_US
dc.contributor.authorBreuer, Gen_US
dc.contributor.authorFischer, Ren_US
dc.date.accessioned2012-08-08T08:48:58Z-
dc.date.available2012-08-08T08:48:58Z-
dc.date.issued2001en_US
dc.identifier.citationJournal Of Virological Methods, 2001, v. 91 n. 2, p. 139-147en_US
dc.identifier.issn0166-0934en_US
dc.identifier.urihttp://hdl.handle.net/10722/157331-
dc.description.abstractGlutathione S-transferase (GST) fusion proteins are used frequently for investigating protein-protein and protein-DNA interactions. The present study demonstrates that the use of GST fusion proteins caused false positives during selection of phage-displayed single-chain antibody fragments (scFvs) specific for three domains of the movement protein (NSM) of tomato spotted wilt virus (TSWV). To identify and exclude the false positives when using GST as a fusion partner linked to the antigen of interest, indirect phage enzyme-linked immunosorbent assay (ELISA) was compared with capture phage ELISA. Of 210 enriched phage clones, indirect phage ELISA identified 106 clones specific for binding to GST-domain fusions but not to GST. In contrast, using capture phage ELISA, all 106 selected clones were identified as false positives, reacting with the GST fusion proteins and GST. This was confirmed by characterization of soluble scFv antibodies. The data indicate that GST fusion proteins seem unsuitable for screening of phage-displayed antibody fragments and it is essential to use capture phage ELISA, instead of the indirect phage ELISA used commonly to exclude false positives in characterization of selected clones with GST fusion proteins. © 2001 Elsevier Science B.V.en_US
dc.languageengen_US
dc.publisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/jvirometen_US
dc.relation.ispartofJournal of Virological Methodsen_US
dc.subject.meshAnimalsen_US
dc.subject.meshAntibodies, Viral - Analysis - Immunologyen_US
dc.subject.meshBacteriophagesen_US
dc.subject.meshBinding Sitesen_US
dc.subject.meshChickensen_US
dc.subject.meshCloning, Molecular - Methodsen_US
dc.subject.meshEnzyme-Linked Immunosorbent Assay - Methodsen_US
dc.subject.meshFalse Positive Reactionsen_US
dc.subject.meshGlutathione Transferase - Genetics - Immunologyen_US
dc.subject.meshImmunoglobulin Fragments - Analysis - Immunologyen_US
dc.subject.meshPeptide Libraryen_US
dc.subject.meshPlant Viral Movement Proteinsen_US
dc.subject.meshProtein Bindingen_US
dc.subject.meshRecombinant Fusion Proteins - Genetics - Immunology - Metabolismen_US
dc.subject.meshSolubilityen_US
dc.subject.meshTospovirus - Immunologyen_US
dc.subject.meshViral Proteins - Genetics - Immunologyen_US
dc.titleGST fusion proteins cause false positives during selection of viral movement protein specific single chain antibodiesen_US
dc.typeArticleen_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1016/S0166-0934(00)00262-7en_US
dc.identifier.pmid11164495-
dc.identifier.scopuseid_2-s2.0-0035146679en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0035146679&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume91en_US
dc.identifier.issue2en_US
dc.identifier.spage139en_US
dc.identifier.epage147en_US
dc.identifier.isiWOS:000166649900005-
dc.publisher.placeNetherlandsen_US

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