Epitope-selected monospecific antibodies to recombinant antigens from Toxoplasma gondii reacted with dense granules of tachyzoites.
Epitope-selected monospecific antibodies were applied to investigate the localization of antigenic molecules in Toxoplasma gondii by immunoelectron microscopy. Eighty cDNA clones encoding antigenic polypeptides were immunoscreened from lambda gt11 expression library with T. gondii infected mouse sera. Twenty different clones with no crossreactivity were selected from eighty clones. Monospecific antibodies to antigens derived from respective cDNA clones extracted from infected mouse sera by the epitope selection method were used in Western blot analysis and immunoelectron microscopy. Eleven antigens were detected with epitope-selected antibodies in lysates of T. gondii tachyzoites. Five of the antigens with molecular weights of 60, 40, 35, 28, and 27 KD were localized in the dense granules. Monospecific antibodies purified by the epitope selection method were useful for investigating the localization of antigens without preparation of a monoclonal antibody from a hybridoma.
Matsuura
T
T
Department of Parasitology, Shinshu University School of Medicine, Nagano, Japan.
Tegoshi
T
T
Furuta-Matsuura
M
M
Sugane
K
K
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000953:Antigens, Protozoan / Q000276:immunology*; D015153:Blotting, Western; D003429:Cross Reactions; D004247:DNA / Q000276:immunology; D000939:Epitopes / Q000276:immunology*; D016253:Microscopy, Immunoelectron; D014122:Toxoplasma / Q000276:immunology* / Q000648:ultrastructure
D000911:Antibodies, Monoclonal; D000953:Antigens, Protozoan; D000939:Epitopes; D004247:DNA