An enzyme-linked immunosorbent assay for the detection of diacetyl (2,3-butanedione)

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

An enzyme-linked immunosorbent assay for the detection of diacetyl (2,3-butanedione). / Marri, Lucia; Jansson, Anita M.; Christensen, Caspar Elo; Hindsgaul, Ole.

In: Analytical Biochemistry, Vol. 535, 2017, p. 12-18.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Marri, L, Jansson, AM, Christensen, CE & Hindsgaul, O 2017, 'An enzyme-linked immunosorbent assay for the detection of diacetyl (2,3-butanedione)', Analytical Biochemistry, vol. 535, pp. 12-18. https://doi.org/10.1016/j.ab.2017.07.021

APA

Marri, L., Jansson, A. M., Christensen, C. E., & Hindsgaul, O. (2017). An enzyme-linked immunosorbent assay for the detection of diacetyl (2,3-butanedione). Analytical Biochemistry, 535, 12-18. https://doi.org/10.1016/j.ab.2017.07.021

Vancouver

Marri L, Jansson AM, Christensen CE, Hindsgaul O. An enzyme-linked immunosorbent assay for the detection of diacetyl (2,3-butanedione). Analytical Biochemistry. 2017;535:12-18. https://doi.org/10.1016/j.ab.2017.07.021

Author

Marri, Lucia ; Jansson, Anita M. ; Christensen, Caspar Elo ; Hindsgaul, Ole. / An enzyme-linked immunosorbent assay for the detection of diacetyl (2,3-butanedione). In: Analytical Biochemistry. 2017 ; Vol. 535. pp. 12-18.

Bibtex

@article{e91b35c9244c46e28028d251a0f45a22,

title = "An enzyme-linked immunosorbent assay for the detection of diacetyl (2,3-butanedione)",

abstract = "Diacetyl (2,3-butanedione) is an important metabolic marker of several cancers, as well as an important off-flavour component produced during fermentation. As a small molecule in a complex mixture with many other analytes, existing methods for identification and quantitation of diacetyl invariably involves a chromatographic separation step followed by signal integration with an appropriate stoichiometric detector. Here we demonstrate that the chemical reaction of diacetyl with a 1,2-phenylenediamine derivative yields a chemical adduct, 1,4-quinoxaline which can be conjugated on BSA. The BSA-diacetyl adduct can be used to select an adduct-specific monoclonal antibody in a Fab-format from a 45-billion member phage-display library. The availability of this antibody allowed the development of an enzyme-linked immunosorbent assay for diacetyl, based on the 1,4-quinoxaline competition for the antibodies with the diacetyl adduct immobilized on the plate. The described ELISA assay can detect the captured diacetyl in micromolar concentrations, both in water samples and in cell culture medium.",

keywords = "Animals, Cattle, Diacetyl/analysis, Enzyme-Linked Immunosorbent Assay, Molecular Structure, Serum Albumin, Bovine/chemistry",

author = "Lucia Marri and Jansson, {Anita M.} and Christensen, {Caspar Elo} and Ole Hindsgaul",

year = "2017",

doi = "10.1016/j.ab.2017.07.021",

language = "English",

volume = "535",

pages = "12--18",

journal = "Analytical Biochemistry",

issn = "0003-2697",

publisher = "Elsevier",

}

RIS

TY - JOUR

T1 - An enzyme-linked immunosorbent assay for the detection of diacetyl (2,3-butanedione)

AU - Marri, Lucia

AU - Jansson, Anita M.

AU - Christensen, Caspar Elo

AU - Hindsgaul, Ole

PY - 2017

Y1 - 2017

N2 - Diacetyl (2,3-butanedione) is an important metabolic marker of several cancers, as well as an important off-flavour component produced during fermentation. As a small molecule in a complex mixture with many other analytes, existing methods for identification and quantitation of diacetyl invariably involves a chromatographic separation step followed by signal integration with an appropriate stoichiometric detector. Here we demonstrate that the chemical reaction of diacetyl with a 1,2-phenylenediamine derivative yields a chemical adduct, 1,4-quinoxaline which can be conjugated on BSA. The BSA-diacetyl adduct can be used to select an adduct-specific monoclonal antibody in a Fab-format from a 45-billion member phage-display library. The availability of this antibody allowed the development of an enzyme-linked immunosorbent assay for diacetyl, based on the 1,4-quinoxaline competition for the antibodies with the diacetyl adduct immobilized on the plate. The described ELISA assay can detect the captured diacetyl in micromolar concentrations, both in water samples and in cell culture medium.

AB - Diacetyl (2,3-butanedione) is an important metabolic marker of several cancers, as well as an important off-flavour component produced during fermentation. As a small molecule in a complex mixture with many other analytes, existing methods for identification and quantitation of diacetyl invariably involves a chromatographic separation step followed by signal integration with an appropriate stoichiometric detector. Here we demonstrate that the chemical reaction of diacetyl with a 1,2-phenylenediamine derivative yields a chemical adduct, 1,4-quinoxaline which can be conjugated on BSA. The BSA-diacetyl adduct can be used to select an adduct-specific monoclonal antibody in a Fab-format from a 45-billion member phage-display library. The availability of this antibody allowed the development of an enzyme-linked immunosorbent assay for diacetyl, based on the 1,4-quinoxaline competition for the antibodies with the diacetyl adduct immobilized on the plate. The described ELISA assay can detect the captured diacetyl in micromolar concentrations, both in water samples and in cell culture medium.

KW - Animals

KW - Cattle

KW - Diacetyl/analysis

KW - Enzyme-Linked Immunosorbent Assay

KW - Molecular Structure

KW - Serum Albumin, Bovine/chemistry

U2 - 10.1016/j.ab.2017.07.021

DO - 10.1016/j.ab.2017.07.021

M3 - Journal article

C2 - 28739133

VL - 535

SP - 12

EP - 18

JO - Analytical Biochemistry

JF - Analytical Biochemistry

SN - 0003-2697

ER -

ID: 200827140

Department of Biology