Partial analysis of the flagellar antigenic determinant recognized by a monoclonal antibody to Clostridium tyrobutyricum.
In order to count Clostridium tyrobutyricum spores in milk after membrane filtration, murine 21E7-B12 monoclonal antibody was produced. Elution of the monoclonal antibody from this antigen, the flagellar filament protein, by carbohydrate ligands was used to study the epitope structure. A competitive elution of an anti-dextran monoclonal antibody by carbohydrate ligands served as a control in order to validate the immunological tool applied to flagellin epitope study. The carbohydrate moiety of flagellin contained D-glucose and N-acetyl-glucosamine in a molar ration of 11:1 as determined by gas-liquid chromatography and 2 low-abundancy unidentified compounds. In ELISA, D-glucose and N-acetyl-glucosamine did not dissociate the antibody-flagellin complex contrary to maltose, maltotriose, maltotetraose and maltopentaose. The efficiency of elution increased from the dimer to the pentamer and became nil for maltohexaose and maltoheptaose. The fact that the hexamer and heptamer could not react with the 21E7-B12 monoclonal antibody could be explained by a drastic conformational change. The over-all stretched maltopentaose switch to a helical-shaped maltoheptaose which could not fit the 21E7-B12 monoclonal antibody antigen-combining site. Thus, flagellin epitope may contain alpha (1-->4) linked glucose residues plus either N-actyl-glucosamine or an unidentified compound that maintain it in an extended shape.
Bédouet
L
L
Université de Bretagne Occidentale, Institut Universitaire de Technologie de Quimper, Laboratoire Universitaire de Microbiologie Appliquée de Quimper, France.
Arnold
F
F
Robreau
G
G
Batina
P
P
Talbot
F
F
Malcoste
R
R
D000911:Antibodies, Monoclonal; D002236:Carbohydrate Conformation; D003013:Clostridium / Q000737:chemistry / Q000254:growth & development / Q000276:immunology*; D000939:Epitopes / Q000737:chemistry* / Q000276:immunology; D005408:Flagellin / Q000737:chemistry*; D009005:Monosaccharides / Q000737:chemistry*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D009005:Monosaccharides; D005408:Flagellin