title: Double-antibody sandwich enzyme-linked immunosorbent assay for cellobiohydrolase I.
abstract: 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 mug/ml.
authors.affiliation: Department of Biochemistry and Microbiology, Cook College, Rutgers University, P. O. Box 231, New Brunswick, New Jersey 08903.
authors.forename: F J
authors.initials: FJ
authors.lastname: Riske
authors.forename: D E
authors.initials: DE
authors.lastname: Eveleigh
authors.forename: J D
authors.initials: JD
authors.lastname: Macmillan