Production of a monoclonal antibody specific to Escherichia coli H33 flagellin by using predetermined polypeptides.
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.
Seah
J N
JN
Animal Health Biotechnology, Institute of Molecular Agrobiology, 1 Research Link, The National University of Singapore, Singapore, 117604.
Kwang
J
J
D000595:Amino Acid Sequence; D000907:Antibodies, Bacterial / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D003001:Cloning, Molecular; D018604:Epitope Mapping; D029968:Escherichia coli Proteins / Q000276:immunology*; D005408:Flagellin / Q000276:immunology*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000276:immunology; D020539:Sequence Analysis, Protein; D017386:Sequence Homology, Amino Acid; D012703:Serotyping
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D029968:Escherichia coli Proteins; C450848:H33 flagellin, E coli; D010446:Peptide Fragments; D005408:Flagellin