Development of polyclonal antibodies against Fusarium verticillioides exoantigens
Exoantigens from Fusarium verticillioides strains were analysed by polyacrylamide gel electrophoresis (SDS-PAGE), enzyme-linked immunosorbent assay (ELISA) and Western blot and were used for production of polyclonal antibodies. Rabbits were immunized with exoantigens from strains 97K, 162A and 113F. Polyclonal antibodies against F. verticillioides 97K exoantigen showed the highest titre in ELISA (1:12.800) and were used in further studies. The cross-reactivity of anti-97K antibodies with 14 species from 7 other fungal genera was low, ranging from 4.5–14.7%; the highest reactivities (51.2–76.0%) were observed in Fusarium genera. The detection limit of exoantigen by indirect competitive ELISA was 0.27 µg/ml. Western blot analysis showed two immunodominant antigens (67 kDa and 113 kDa) common to all the F. verticillioides strains analysed. The results suggest that polyclonal antibodies raised against exoantigens from F. verticillioides 97K are promising for Fusarium sp. immunodetection.
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enzyme-linked immunosorbent assay;
Document Type: Research Article
Department of Biochemistry and Biotechnology, State University of Londrina, Paraná, Brazil
Department of Pathological Sciences, State University of Londrina, Paraná, Brazil
Institute of Food Technology, Campinas, São Paulo, Brazil
Kobe Institute of Health – Department of Food Chemistry Chuo-ku, Kobe, Japan
Faculty of Pharmaceutical Sciences, Science University of Tokyo, Ichigaya, Tokyo, Japan
Department of Food and Drug Technology, State University of Londrina, Paraná, Brazil
March 1, 2006
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