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Quantitation ofAcidothermus cellulolyticus E1 endoglucanase andThermomonospora fusca E3 exoglucanase using enzyme-linked immunosorbent assay (ELISA)

  • Session 2 Past, Present, and Emerging Concepts in Applied Biological Research
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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 forAcidothermus cellulolyticus E1 endoglucanase. Likewise, a polyclonal rabbit serum (Ab684) was used to develop an ELISA specific forThermomonospora 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 ofStreptomyces lividans strain TK24 in which the corresponding genes have been cloned and expressed.

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Authors and Affiliations

  1. Alternative Fuels Division, National Renewable Energy Laboratory, Applied Biological Sciences Branch, 80401, Golden, CO

    Rafael A. Nieves, Yat-Chen Chou, Michael E. Himmel & Steven R. Thomas

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  1. Rafael A. Nieves
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  2. Yat-Chen Chou
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  3. Michael E. Himmel
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  4. Steven R. Thomas
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Nieves, R.A., Chou, YC., Himmel, M.E. et al. Quantitation ofAcidothermus cellulolyticus E1 endoglucanase andThermomonospora fusca E3 exoglucanase using enzyme-linked immunosorbent assay (ELISA). Appl Biochem Biotechnol 51, 211–223 (1995). https://doi.org/10.1007/BF02933425

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