Quantitation of Acidothermus cellulolyticus E1 endoglucanase and Thermomonospora fusca E3 exoglucanase using enzyme-linked immunosorbent assay (ELISA)

Abstract

Two distinct quantitative indirect ELISAs were developed to determine the concentration of recombinant cellulase enzymes in culture filtrates. A monoclonal antibody (E1P7) was used as the primary antibody in developing an ELISA specific for Acidothermus cellulolyticus E1 endoglucanase. Likewise, a polyclonal rabbit serum (Ab684) was used to develop an ELISA specific for Thermomonospora fusca E3 exoglucanase. Dose-response curves indicated a dynamic range for both assays between 0.01 and 0.08 μg/mL (1-8 ng/assay) when purified enzymes were used as standards. These assays have been used to estimate concentrations of secreted recombinant E1 and/or E3 in culture supernatants of Streptomyces lividans strain TK24 in which the corresponding genes have been cloned and expressed. © 1995 Humana Press Inc.