A single chain Fv antibody displayed on phage surface recognises conformational group-specific epitope of bluetongue virus

• Published in: Journal of virological methods Vol. 91; no. 2; pp. 203 - 207
• Main Authors: Nagesha, H.S, Wang, L.-F, Shiell, B, Beddome, G, White, J.R, Irving, R.A
• Format: Journal Article
• Language: English
• Published: London Elsevier B.V 01.02.2001; Amsterdam Elsevier; New York, NY Elsevier Science B.V
• Subjects: Amino Acid Sequence; Antibodies, Monoclonal - biosynthesis; Antibodies, Monoclonal - genetics; Antibodies, Monoclonal - immunology; Antibodies, Viral - genetics; Antibodies, Viral - immunology; Antibodies, Viral - isolation & purification; Antibody Specificity; Bacteriophages; Base Sequence; Binding Sites, Antibody; Biological and medical sciences; Bluetongue virus; Bluetongue virus - immunology; Brief Report; Cloning, Molecular; Coat protein; Codon, Terminator; DNA, Viral; Epitopes, B-Lymphocyte - immunology; Filamentous phage; Fundamental and applied biological sciences. Psychology; Immunoglobulin Fragments - genetics; Immunoglobulin Fragments - immunology; Immunoglobulin Fragments - isolation & purification; Immunoglobulin Variable Region - genetics; Immunoglobulin Variable Region - immunology; Microbiology; Molecular Sequence Data; Peptide Library; ScFv; Sequence Analysis, DNA; Viral Core Proteins - immunology; Coat protein; Filamentous phage; ScFv; Bluetongue virus; Antigenic determinant; Hybridoma; Monoclonal antibody; Reoviridae; Virus; Phage display; Cell line; Gene; Orbivirus; Single chain antibody; Phage; Conformation
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/S0166-0934(00)00266-4

A single chain fragment variable (scFv) antibody gene was isolated from hybridoma cell line secreting monoclonal antibody (MAb) 20E9 that recognises bluetongue virus (BTV) VP7. DNA fragments encoding variable regions of heavy and light chains were amplified by RT-PCR and library of scFv was constructed in phage vector. Two scFv clones that were selected showed specific reactivity with conformational epitope VP7. The N-terminal 22 amino acid residues of 20E9 light chain were identical to that deduced from scFv DNA sequence. An in-frame TAG stop codon was found in the coding sequence and its potential role in regulating the expression and stability of scFv in phage is discussed.