Production of a monoclonal antibody specific to Escherichia coli H33 flagellin by using predetermined polypeptides

• Published in: Archives of microbiology 2002-02, Vol.177 (2), p.192-196
• Main Authors: Seah, J, Kwang, J
• Format: Article
• Language: English
• Subjects: agglutination; Amino Acid Sequence; amino acids; Antibodies, Bacterial - immunology; Antibodies, Monoclonal - immunology; Antibody Specificity; antiserum; Bacteriology; Biological and medical sciences; Cloning, Molecular; cross reaction; Epitope Mapping; epitopes; Escherichia coli; Escherichia coli Proteins - immunology; flagellin; Flagellin - immunology; Fundamental and applied biological sciences. Psychology; genes; immunoglobulin G; mice; Microbiology; Molecular Sequence Data; monoclonal antibodies; Peptide Fragments - immunology; polypeptides; Sequence Analysis, Protein; Sequence Homology, Amino Acid; serotypes; Serotyping; Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies
• ISSN: 0302-8933; 1432-072X
• DOI: 10.1007/s00203-001-0374-2

The Escherichia coli H serogroup is determined by flagellin, which has both H-type-specific and cross-reactive epitopes. The cross-reactive epitopes are responsible for the cross-reaction found in agglutination. To identify the specific epitope in H33 flagellin, the H33 flagellin gene was sequenced and the encoded central variable region (CVR) was determined. Four overlapping fragments of the CVR were prepared and their specificity was verified using different H-type antisera. Short fragments carrying potential H-type-specific determinants were selected, and monoclonal antibodies (MAbs) against these fragments were prepared. A murine MAb of subtype IgG1 showing specificity to H33 flagellin was produced. The epitope of the MAb was mapped to amino acid residues 250–260.