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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
- Source :
- Clinical and Experimental Immunology. 125:94-101
- Publication Year :
- 2001
- Publisher :
- Oxford University Press (OUP), 2001.
-
Abstract
- SummaryWe 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.
- Subjects :
- Phage display
Recombinant Fusion Proteins
Molecular Sequence Data
Immunology
Immunoglobulin Variable Region
Dot blot
chemical and pharmacologic phenomena
medicine.disease_cause
Viral Envelope Proteins
Antibody Specificity
Neutralization Tests
Peptide Library
Chlorocebus aethiops
medicine
Animals
Humans
Immunology and Allergy
Single-chain variable fragment
Amino Acid Sequence
Mononegavirales
Antigens, Viral
Immunoglobulin Fragments
Vero Cells
Lyssavirus
Glycoproteins
biology
Rabies virus
Immunity to Infection
respiratory system
Rhabdoviridae
biology.organism_classification
Fragment crystallizable region
Virology
Molecular biology
Solubility
Immunoglobulin G
Subjects
Details
- ISSN :
- 13652249 and 00099104
- Volume :
- 125
- Database :
- OpenAIRE
- Journal :
- Clinical and Experimental Immunology
- Accession number :
- edsair.doi.dedup.....8ce4abfeb62f82f34a25e8776bcf76ec
- Full Text :
- https://doi.org/10.1046/j.1365-2249.2001.01515.x