Production of monoclonal antibodies specific for the i and 1,2 flagellar antigens of Salmonella typhimurium and characterization of their respective epitopes

• Published in: Applied and Environmental Microbiology 1998-12, Vol.64 (12), p.5033-5038
• Main Authors: Vries, N. de (Utrecht University, Utrecht.), Zwaagstra, K.A, Huis in't Veld, J.H.J, Knapen, F. van, Zijderveld, F.G. van, Kusters, J.G
• Format: Article
• Language: English
• Subjects: Antibodies; Antibodies, Monoclonal; Antibody Specificity; ANTIGENE RECOMBINANT; ANTIGENOS RECOMBINANTES; Antigens, Bacterial - genetics; Antigens, Bacterial - immunology; Bacterial Proteins; Bacteriological methods and techniques used in bacteriology; Bacteriology; Biological and medical sciences; CHICKENS; Cloning, Molecular; DNA Primers; ELISA; Epitopes - analysis; EXPERIMENTACION IN VIVO; EXPERIMENTAL INFECTION; EXPERIMENTATION IN VIVO; Flagellin - genetics; Flagellin - immunology; Food contamination & poisoning; Food Microbiology; Fundamental and applied biological sciences. Psychology; GENBANK/AF045151; Genes; Genes, Bacterial; Humans; Immunology; IN VIVO EXPERIMENTATION; INFECCION EXPERIMENTAL; INFECTION EXPERIMENTALE; Microbiology; MOLECULAR SEQUENCE DATA; NUCLEOTIDE SEQUENCE; POLLO; Polymerase Chain Reaction; POULET; RECOMBINANT ANTIGENS; Recombinant Proteins - immunology; Reproducibility of Results; Restriction Mapping; Salmonella; Salmonella - immunology; SALMONELLA TYPHIMURIUM; Salmonella typhimurium - genetics; Salmonella typhimurium - immunology; Salmonella typhimurium - isolation & purification; SECUENCIA NUCLEOTIDICA; SEQUENCE NUCLEOTIDIQUE; TEST ELISA
• ISSN: 0099-2240; 1098-5336
• DOI: 10.1128/aem.64.12.5033-5038.1998

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