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Development of polyclonal antibodies against Fusarium verticillioides exoantigens

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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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Keywords: ELISA; Fusarium sp; enzyme-linked immunosorbent assay; exoantigens; immunodetection

Document Type: Research Article

Affiliations: 1: Department of Biochemistry and Biotechnology, State University of Londrina, Paraná, Brazil 2: Department of Pathological Sciences, State University of Londrina, Paraná, Brazil 3: Institute of Food Technology, Campinas, São Paulo, Brazil 4: Kobe Institute of Health – Department of Food Chemistry Chuo-ku, Kobe, Japan 5: Faculty of Pharmaceutical Sciences, Science University of Tokyo, Ichigaya, Tokyo, Japan 6: Department of Food and Drug Technology, State University of Londrina, Paraná, Brazil

Publication date: 2006-03-01

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