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. 1990 Nov;56(11):3261–3265. doi: 10.1128/aem.56.11.3261-3265.1990

Double-Antibody Sandwich Enzyme-Linked Immunosorbent Assay for Cellobiohydrolase I

Frank J Riske 1,†, Douglas E Eveleigh 1, James D Macmillan 1,*
PMCID: PMC184939  PMID: 16348332

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 β-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 μg/ml.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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