Visualization of Trichoderma reesei cellobiohydrolase I and endoglucanase I on aspen cellulose by using monoclonal antibody-colloidal gold conjugates

Monoclonal antibodies (MAbs) specific for cellobiohydrolase I (CBH 1) and endoglucanase I (EG 1) were conjugated to 10- and 15-nm colloidal gold particles, respectively. The binding of CBH I and EG I was visualized by utilizing the MAb-colloidal gold probes. The visualization procedure involved immo...

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Veröffentlicht in:Applied and Environmental Microbiology 1991-11, Vol.57 (11), p.3163-3170
Hauptverfasser: Nieves, R.A. (Solar Energy Research Institute, Golden, CO), Ellis, R.P, Todd, R.J, Johnson, T.J.A, Grohmann, K, Himmel, M.E
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Sprache:eng
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Zusammenfassung:Monoclonal antibodies (MAbs) specific for cellobiohydrolase I (CBH 1) and endoglucanase I (EG 1) were conjugated to 10- and 15-nm colloidal gold particles, respectively. The binding of CBH I and EG I was visualized by utilizing the MAb-colloidal gold probes. The visualization procedure involved immobilization of cellulose microfibrils on copper electron microscopy grids, incubation of the cellulose-coated grids with cellulase(s), binding of MAb-colloidal gold conjugates to cellulase(s), and visualization via transmission electron microscopy. CBH I was seen bound to apparent crystalline cellulose as well as apparent amorphous cellulose. EG I was seen bound extensively to apparent amorphous cellulose with minimal binding to crystalline cellulose
ISSN:0099-2240
1098-5336
DOI:10.1128/AEM.57.11.3163-3170.1991