Selection of single chain variable fragments (scFv) against the glycoprotein antigen of the rabies virus from a human synthetic scFv phage display library and their fusion with the Fc region of human IgG1.

2.50
Hdl Handle:
http://hdl.handle.net/10541/85755
Title:
Selection of single chain variable fragments (scFv) against the glycoprotein antigen of the rabies virus from a human synthetic scFv phage display library and their fusion with the Fc region of human IgG1.
Authors:
Ray, K; Embleton, Jim; Jailkhani, B L; Bhan, M K; Kumar, R
Abstract:
We have prepared human recombinant antibody molecules against the glycoprotein antigen of the rabies virus (GPRV) based on the single chain variable fragment (scFv) format. Anti-GPRV scFvs were selected from a human synthetic scFv phage display library with a repertoire of approximately 109 specificities. After three rounds of selection against the PV11 strain of the virus, 40% of the clones tested recognized the rabies antigen. Of the 20 positive clones that were sequenced, five distinct sequences were identified. These distinct scFvs were cloned into a mammalian expression vector carrying the human IgG1 Fc region. The specificity of the resulting scFv-Fc molecules for GPRV was established by ELISA, dot blot and western blot analyses and membrane immunofluorescence. Two of the scFv-Fc fusion proteins neutralized the PV11 strain in a standard in vivo neutralization assay where the virus was incubated with the scFv-Fc molecules before intracranial inoculation in mice. These anti-GPRV scFv-Fc molecules have the potential to be used as an alternative to the presently available HRIG, for use in post-exposure preventive treatment.
Affiliation:
Department of Paediatrics, All India Institute of Medical Sciences, Ansari Nagar, New Delhi, India.
Citation:
Selection of single chain variable fragments (scFv) against the glycoprotein antigen of the rabies virus from a human synthetic scFv phage display library and their fusion with the Fc region of human IgG1. 2001, 125 (1):94-101 Clin. Exp. Immunol.
Journal:
Clinical and Experimental Immunology
Issue Date:
Jul-2001
URI:
http://hdl.handle.net/10541/85755
PubMed ID:
11472431
Type:
Article
Language:
en
ISSN:
0009-9104
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorRay, Ken
dc.contributor.authorEmbleton, Jimen
dc.contributor.authorJailkhani, B Len
dc.contributor.authorBhan, M Ken
dc.contributor.authorKumar, Ren
dc.date.accessioned2009-11-10T10:18:21Z-
dc.date.available2009-11-10T10:18:21Z-
dc.date.issued2001-07-
dc.identifier.citationSelection of single chain variable fragments (scFv) against the glycoprotein antigen of the rabies virus from a human synthetic scFv phage display library and their fusion with the Fc region of human IgG1. 2001, 125 (1):94-101 Clin. Exp. Immunol.en
dc.identifier.issn0009-9104-
dc.identifier.pmid11472431-
dc.identifier.urihttp://hdl.handle.net/10541/85755-
dc.description.abstractWe have prepared human recombinant antibody molecules against the glycoprotein antigen of the rabies virus (GPRV) based on the single chain variable fragment (scFv) format. Anti-GPRV scFvs were selected from a human synthetic scFv phage display library with a repertoire of approximately 109 specificities. After three rounds of selection against the PV11 strain of the virus, 40% of the clones tested recognized the rabies antigen. Of the 20 positive clones that were sequenced, five distinct sequences were identified. These distinct scFvs were cloned into a mammalian expression vector carrying the human IgG1 Fc region. The specificity of the resulting scFv-Fc molecules for GPRV was established by ELISA, dot blot and western blot analyses and membrane immunofluorescence. Two of the scFv-Fc fusion proteins neutralized the PV11 strain in a standard in vivo neutralization assay where the virus was incubated with the scFv-Fc molecules before intracranial inoculation in mice. These anti-GPRV scFv-Fc molecules have the potential to be used as an alternative to the presently available HRIG, for use in post-exposure preventive treatment.en
dc.language.isoenen
dc.subject.meshAmino Acid Sequence-
dc.subject.meshAnimals-
dc.subject.meshAntibody Specificity-
dc.subject.meshAntigens, Viral-
dc.subject.meshCercopithecus aethiops-
dc.subject.meshGlycoproteins-
dc.subject.meshHumans-
dc.subject.meshImmunoglobulin Fragments-
dc.subject.meshImmunoglobulin G-
dc.subject.meshImmunoglobulin Variable Region-
dc.subject.meshMolecular Sequence Data-
dc.subject.meshNeutralization Tests-
dc.subject.meshPeptide Library-
dc.subject.meshRabies virus-
dc.subject.meshRecombinant Fusion Proteins-
dc.subject.meshSolubility-
dc.subject.meshVero Cells-
dc.subject.meshViral Envelope Proteins-
dc.titleSelection of single chain variable fragments (scFv) against the glycoprotein antigen of the rabies virus from a human synthetic scFv phage display library and their fusion with the Fc region of human IgG1.en
dc.typeArticleen
dc.contributor.departmentDepartment of Paediatrics, All India Institute of Medical Sciences, Ansari Nagar, New Delhi, India.en
dc.identifier.journalClinical and Experimental Immunologyen

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