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

• Published in: Clinical and experimental immunology 2001-07, Vol.125 (1), p.94-101
• Main Authors: Ray, K., Embleton, M. J., Jailkhani, B. L., Bhan, M. K., Kumar, R.
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Antibody Specificity - immunology; Antigens, Viral - immunology; Biological and medical sciences; Chlorocebus aethiops; Fundamental and applied biological sciences. Psychology; glycoprotein; Glycoproteins - immunology; human scFv; Human viral diseases; Humans; Immunity to Infection; Immunoglobulin Fragments - biosynthesis; Immunoglobulin Fragments - genetics; Immunoglobulin Fragments - immunology; Immunoglobulin G - genetics; Immunoglobulin G - immunology; immunoglobulin G1; Immunoglobulin Variable Region - biosynthesis; Immunoglobulin Variable Region - genetics; Immunoglobulin Variable Region - immunology; Infectious diseases; Medical sciences; Microbiology; Molecular Sequence Data; Neutralization Tests; neutralizing; Peptide Library; phage display; rabies; Rabies virus; Rabies virus - immunology; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - immunology; scFv‐Fc; Solubility; Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies; Vero Cells; Viral diseases; Viral diseases of the nervous system; Viral Envelope Proteins - immunology; Virology
• ISSN: 0009-9104; 1365-2249
• DOI: 10.1046/j.1365-2249.2001.01515.x

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.