Production of monoclonal antibodies specific for the i and 1,2 flagellar antigens of Salmonella typhimurium and characterization of their respective epitopes.
Salmonella typhimurium expresses two antigenically distinct flagellins, each containing a different H antigen (i and 1,2), the combination of which is highly specific for this serotype. In this study, overlapping recombinant flagellin fragments were constructed from the fliC (H:i) and fljB (H:1,2) flagellin genes, and the expression products were tested for binding to H antigen-specific monoclonal and polyclonal antibodies. A minimal area, 86 amino acids for H:i and 102 amino acids for H:1,2, located in the central variable domain of each flagellin was required for the binding of serotype-specific antibodies, providing further evidence for the presence of a discontinuous H epitope. Two peptides comprising these areas were shown to be highly suitable for application as antigens in an enzyme-linked immunosorbent assay detecting S. typhimurium-specific antibody.
de Vries
N
N
Department of the Science of Food of Animal Origin, The Netherlands.
Zwaagstra
K A
KA
Huis in't Veld
J H
JH
van Knapen
F
F
van Zijderveld
F G
FG
Kusters
J G
JG
D000906:Antibodies; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology; D001426:Bacterial Proteins; D003001:Cloning, Molecular; D017931:DNA Primers; D000939:Epitopes / Q000032:analysis; D005408:Flagellin / Q000235:genetics / Q000276:immunology*; D005798:Genes, Bacterial; D006801:Humans; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000276:immunology; D015203:Reproducibility of Results; D015183:Restriction Mapping; D012475:Salmonella / Q000276:immunology; D012486:Salmonella typhimurium / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification
D000906:Antibodies; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D017931:DNA Primers; D000939:Epitopes; C094427:FljB protein, Salmonella; D011994:Recombinant Proteins; D005408:Flagellin