Sequence analysis, expression, and paratope characterization of a single-chain Fv fragment for the eukaryote ribosomal P proteins

• Published in: Biochemical and biophysical research communications 2003-02, Vol.301 (4), p.819-824
• Main Authors: López Bergami, Pablo, Mateos, Pablo, Hoebeke, Johan, Levin, Mariano Jorge, Baldi, Alberto
• Format: Article
• Language: English
• Subjects: Amino Acid Motifs; Amino Acid Sequence; Animals; Antibodies, Monoclonal - chemistry; Antibodies, Monoclonal - genetics; Autoantibody; Binding Sites, Antibody - genetics; DNA - genetics; Escherichia coli; Genes, Immunoglobulin; Immunoglobulin Variable Region - chemistry; Immunoglobulin Variable Region - genetics; Mice; Models, Molecular; Molecular Sequence Data; Paratope–epitope modelling; Protozoan Proteins - immunology; Ribosomal P protein; Ribosomal Proteins - immunology; Sequence Homology, Amino Acid; Single-chain Fv fragment; Trypanosoma cruzi; Trypanosoma cruzi - immunology
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/S0006-291X(02)03074-7

The variable genes of monoclonal antibody (mAb) B10, specific for the C-terminal region of the eukaryotic ribosomal P protein, have been cloned as a single-chain Fv fragment (scFv) and expressed in Escherichia coli. The primary sequence of the variable regions of the B10 antibody, together with a detailed characterization of the reactive residues of the antigen, allowed the construction of a model of the paratope–epitope interaction, giving a first insight into the binding mechanisms of anti-P autoantibodies to their target peptides. The mAb and scFv could be useful for extensive P protein detection since both recognize the highly conserved motif DDxGF.