Monoclonal antibodies (IF3(1), 4G3(14), 4G2(4), and 2B6(24)) produced against cell wall protein .... Ophiostoma piceae and G. ~.oserrnl cell walls were isolated and cell .... Ophiostoma piceaeN. 3871. 100.0. 100.0. 100.0. 100.0. Alternaria alternata. 2G .... BSA, 0.1% gelatin, 1% normal goat serum, and 2 mM NaN3 in PBS,.
Production and characterization of monoclonal antibodies to the sapstaining fungus Ophiostoma piceae SRABANI BANERJEE AND JUDYLITTLE Department of Biology, Utlive~sityof Ottawcr, Ottawa, ON KIN 6N5, Canada
MARIACHAN For.ir~tekCatladc~Corp., 800 Montreal Road, Ottawa, ON K l G 3G5, Canada
BRIANT. LUCK Department of Biology, Unive~sityof Otfcrwa, Ottawa, ON KIN 6N5, Canada
Can. J. Microbiol. Downloaded from www.nrcresearchpress.com by Renmin University of China on 06/06/13 For personal use only.
COLETTEBREUIL Forest Products Biotechnolog~~, Faculty o f Forestry, Utziver.sity of British Columbia, Vanco~lver, BC V6T 124, Canada AND
DAVIDL. BROWN' Departrner7t of Biology, University of Ottawa, Ottawa, ON KIN 6N5, Canada Received July 2 1, 1993 Revision received October 26, 1993 Accepted November 1, 1993 BANERJEE, S., LITTLE,J., CHAN,M., LUCK, B.T., BREUIL,C., and BROWN,D.L. 1994. Production and characterization of monoclonal antibodies to the sapstaining fungus Ophiosroma piceae. Can. J. Microbiol. 40: 35-44. A sensitive immunological tool has been developed to detect the sapstaining fungus Ophiosroma piceae 3871, which plagues the wood industry. Monoclonal antibodies (IF3(1), 4G3(14), 4G2(4), and 2B6(24)) produced against cell wall protein extracts of this fungus were specific. Specificity was estimated by enzyme linked immunosorbent assay, westem blotting, and light and electron microscopy using the immunogold technique. Electron microscopy revealed gold particles localized on the outer surface of the cell wall. When screened against 24 biological control fungi the antibodies showed pratically no cross-reactivity (hora botulispora, and the antibodies showed less than 5% reactivity with the other fungi. Chemical and enzymatic modification of the antigen revealed that the epitopes recognized by the monoclonal antibodies were glycospecific. Although the antibodies were produced against the cell wall protein extracts of the fungus grown in liquid culture, they also recognized the fungus growing in wood and therefore can be employed to investigate wood colonization by this fungus. piceae, monoclonal antibodies, glycoprotein Key words: O~~lziosronza BANERJEE, S., LITTLE,J., CHAN,M., LUCK,B.T., BREUIL,C., et BROWN, D.L. 1994. Production and characterization of monoclonal antibodies to the sapstaining fungus Ophiostomapiceae. Can. J. Microbiol. 40 : 35-44. Un outil immunologique sensible a CtC dCveloppC pour la dCtection du champignon Ophiosromapiceae 3871, qui colore la s h e et constitue un fleau pour I'industrie du bois. Les anticorps monoclonaux (1F3(1), 4G3(14), 4G2(4) et 2B6(24)), produits contre un extrait de protCines de la paroi cellulaire de ce champignon, ont CtC sptcifiques. La spCcificitC a CtC CvaluCe par Cpreuve ELISA, immunoempreinte, microscopie optique et microscopie Clectronique B I'aide de complexes immunoglobulines-or. La microscopie Clectronique a rCvClC des particules d'or localisees sur la surface externe de la paroi cellulaire. Les anticorps n'ont presque pas montrC de rCactions croisCes (
