Quantitation of Acidothermus cellulolyticus E1 endoglucanase and Thermomonospora fusca E{sub 3} exoglucanase using enzyme-linked immunosorbent assay (ELISA)
- National Renewable Energy Lab., Golden, CO (United States); and others
Two distinct quantitative indirect ELISAs were developed to determine the concentration of recombinant cellulose 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 E{sub 3} exoglucanase. Dose-response curves indicated a dynamic range for both assays between 0.01 and 0.08 {mu}g/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 E{sub 3} in culture supernatants of Streptomyces lividans strain TK24 in which the corresponding genes have been cloned and expressed.
- Research Organization:
- Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
- OSTI ID:
- 150409
- Report Number(s):
- CONF-940526-; TRN: 95:007973-0021
- Resource Relation:
- Conference: 16. symposium on biotechnology for fuels and chemicals, Gatlinburg, TN (United States), 9-13 May 1994; Other Information: PBD: 1995; Related Information: Is Part Of Sixteenth symposium on biotechnology for fuels and chemicals; Davison, B.H. [ed.] [Oak Ridge National Lab., TN (United States)]; Wyman, C.E. [ed.] [National Renewable Energy Lab., Golden, CO (United States)]; PB: 823 p.
- Country of Publication:
- United States
- Language:
- English
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