Quantitation of Acidothermus cellulolyticus E1 Endoglucanase and Thermomonospora fusca E3 Exoglucanase Using Enzyme-Linked Immunosorbent Assay (ELISA)

Rafael A. Nieves; Yat Chen Chou; Michael E. Himmel; Steven R. Thomas

doi:10.1007/BF02933425

Quantitation of Acidothermus cellulolyticus E1 Endoglucanase and Thermomonospora fusca E3 Exoglucanase Using Enzyme-Linked Immunosorbent Assay (ELISA)

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

National Renewable Energy Laboratory

Research output: Contribution to journal › Article › peer-review

12 Scopus Citations

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.

Original language	American English
Pages (from-to)	211-223
Number of pages	13
Journal	Applied Biochemistry and Biotechnology - Part A Enzyme Engineering and Biotechnology
Volume	51-52
Issue number	1
DOIs	https://doi.org/10.1007/BF02933425 https://doi.org/10.1007/BF02933425
State	Published - 1995

NREL Publication Number

NREL/JA-422-6647

Keywords

Acidothermus cellulolyticus
antibodies
Cellulases
ELISA
Thermomonospora fusca

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Nieves, R. A., Chou, Y. C., Himmel, M. E., & Thomas, S. R. (1995). Quantitation of Acidothermus cellulolyticus E1 Endoglucanase and Thermomonospora fusca E3 Exoglucanase Using Enzyme-Linked Immunosorbent Assay (ELISA). Applied Biochemistry and Biotechnology - Part A Enzyme Engineering and Biotechnology, 51-52(1), 211-223. https://doi.org/10.1007/BF02933425, https://doi.org/10.1007/BF02933425

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title = "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.",

keywords = "Acidothermus cellulolyticus, antibodies, Cellulases, ELISA, Thermomonospora fusca",

author = "Nieves, {Rafael A.} and Chou, {Yat Chen} and Himmel, {Michael E.} and Thomas, {Steven R.}",

year = "1995",

doi = "10.1007/BF02933425",

language = "American English",

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pages = "211--223",

journal = "Applied Biochemistry and Biotechnology - Part A Enzyme Engineering and Biotechnology",

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Nieves, RA, Chou, YC , Himmel, ME & Thomas, SR 1995, 'Quantitation of Acidothermus cellulolyticus E1 Endoglucanase and Thermomonospora fusca E3 Exoglucanase Using Enzyme-Linked Immunosorbent Assay (ELISA)', Applied Biochemistry and Biotechnology - Part A Enzyme Engineering and Biotechnology, vol. 51-52, no. 1, pp. 211-223. https://doi.org/10.1007/BF02933425, https://doi.org/10.1007/BF02933425

Quantitation of Acidothermus cellulolyticus E1 Endoglucanase and Thermomonospora fusca E3 Exoglucanase Using Enzyme-Linked Immunosorbent Assay (ELISA). / Nieves, Rafael A.; Chou, Yat Chen ; Himmel, Michael E. et al.
In: Applied Biochemistry and Biotechnology - Part A Enzyme Engineering and Biotechnology, Vol. 51-52, No. 1, 1995, p. 211-223.

Research output: Contribution to journal › Article › peer-review

TY - JOUR

T1 - Quantitation of Acidothermus cellulolyticus E1 Endoglucanase and Thermomonospora fusca E3 Exoglucanase Using Enzyme-Linked Immunosorbent Assay (ELISA)

AU - Nieves, Rafael A.

AU - Chou, Yat Chen

AU - Himmel, Michael E.

AU - Thomas, Steven R.

PY - 1995

Y1 - 1995

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AB - 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.

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Quantitation of Acidothermus cellulolyticus E1 Endoglucanase and Thermomonospora fusca E3 Exoglucanase Using Enzyme-Linked Immunosorbent Assay (ELISA)

Abstract

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Keywords

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