Double-antibody sandwich enzyme-linked immunosorbent assay for cellobiohydrolase I

Journal Article · Thu Nov 01 00:00:00 EST 1990 · Applied and Environmental Microbiology; (USA)

OSTI ID:5912731

Riske, F J; Eveleigh, D E; MacMillan, J D ^[1]

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1. Rutgers Univ., New Brunswick, NJ (USA)

A double-antibody sandwich enzyme-linked immunosorbent assay was developed for quantifying cellobiohydrolase I (CBH I) in crude preparations of the cellulase complex from Trichoderma reesei. The other enzymes (endoglucanase and {beta}-glucosidase) in this complex and other ingredients in culture broth did not interfere with this assay. The antibody configuration that resulted in the highest specificity for the assay of CBH I employed a monoclonal antibody to coat wells in polystyrene plates and peroxidase-labeled polyclonal antibody to detect cellobiohydrolase bound to the immobilized monoclonal antibody. Previously, procedures have not been available for the direct assay of CBH I activity in the presence of the other enzymes in the complex, and current indirect procedures are cumbersome and inaccurate. The direct procedure described here is highly specific for CBH I and useful for quantifying this enzyme in the range of 0.1 to 0.8 {mu}g/ml.

• OSTI ID: 5912731
• Journal Information: Applied and Environmental Microbiology; (USA), Vol. 56:11; ISSN 0099-2240
• Country of Publication: United States
• Language: English

Related Subjects

09 BIOMASS FUELS
CELLULASE
ENZYME IMMUNOASSAY
BIOCONVERSION
MEASURING METHODS
MONOCLONAL ANTIBODIES
SPECIFICITY
TRICHODERMA VIRIDE
ANTIBODIES
BIOASSAY
ENZYMES
EUMYCOTA
FUNGI
GLYCOSYL HYDROLASES
HYDROLASES
IMMUNOASSAY
O-GLYCOSYL HYDROLASES
PLANTS
TRICHODERMA
090900* - Biomass Fuels- Processing- (1990-)