A flow-through fluorescence polarization detection system for measuring GPCR-mediated modulation of cAMP production

J. Kool; A. van Marle; ST Hulscher; M. Selman; D.J. van Iperen; K. van Altena; M.R. Gillard; R.A. Bakker; H. Irth; R. Leurs; N.P.E. Vermeulen

doi:10.1177/1087057107308881

A flow-through fluorescence polarization detection system for measuring GPCR-mediated modulation of cAMP production

J. Kool, A. van Marle, ST Hulscher, M. Selman, D.J. van Iperen, K. van Altena, M.R. Gillard, R.A. Bakker, H. Irth, R. Leurs, N.P.E. Vermeulen

Research output: Contribution to Journal › Article › Academic › peer-review

Abstract

A flow-through fluorescence polarization (FP) detection system that makes use of a novel high-performance liquid chromatography (HPLC) fluorescence detector modified with polarization filters was developed. This flow-through FP detection system was evaluated by using a novel and very cost-effective bioassay for cyclic adenosine monophosphate (cAMP). The bioassay was first evaluated and optimized in an FP plate reader format and subsequently in a flow-through bioassay setup. The principle of the bioassay is based on the competition of cAMP and a fluorescent cAMP derivative for the cAMP binding domain of protein kinase A. cAMP could accurately be determined over a range of 0.8 to 30 pmol/well in the plate reader FP assay and over a range of 0.3 to 50 pmol/well in the flow-through FP assay setup. High Z′ factors (i.e., 0.89 for the plate reader and 0.93 for the flow-through FP cAMP assay, respectively) indicated robust assays. Finally, functional cAMP signaling of the human histamine H

Original language	English
Pages (from-to)	1074-83
Journal	Journal of Biomolecular Screening
Volume	12
Issue number	8
DOIs	https://doi.org/10.1177/1087057107308881
Publication status	Published - 2007

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Kool, J., van Marle, A., Hulscher, ST., Selman, M., van Iperen, D. J., van Altena, K., Gillard, M. R., Bakker, R. A., Irth, H., Leurs, R., & Vermeulen, N. P. E. (2007). A flow-through fluorescence polarization detection system for measuring GPCR-mediated modulation of cAMP production. Journal of Biomolecular Screening, 12(8), 1074-83. https://doi.org/10.1177/1087057107308881

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title = "A flow-through fluorescence polarization detection system for measuring GPCR-mediated modulation of cAMP production",

abstract = "A flow-through fluorescence polarization (FP) detection system that makes use of a novel high-performance liquid chromatography (HPLC) fluorescence detector modified with polarization filters was developed. This flow-through FP detection system was evaluated by using a novel and very cost-effective bioassay for cyclic adenosine monophosphate (cAMP). The bioassay was first evaluated and optimized in an FP plate reader format and subsequently in a flow-through bioassay setup. The principle of the bioassay is based on the competition of cAMP and a fluorescent cAMP derivative for the cAMP binding domain of protein kinase A. cAMP could accurately be determined over a range of 0.8 to 30 pmol/well in the plate reader FP assay and over a range of 0.3 to 50 pmol/well in the flow-through FP assay setup. High Z′ factors (i.e., 0.89 for the plate reader and 0.93 for the flow-through FP cAMP assay, respectively) indicated robust assays. Finally, functional cAMP signaling of the human histamine H",

author = "J. Kool and {van Marle}, A. and ST Hulscher and M. Selman and {van Iperen}, D.J. and {van Altena}, K. and M.R. Gillard and R.A. Bakker and H. Irth and R. Leurs and N.P.E. Vermeulen",

year = "2007",

doi = "10.1177/1087057107308881",

language = "English",

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T1 - A flow-through fluorescence polarization detection system for measuring GPCR-mediated modulation of cAMP production

AU - Kool, J.

AU - van Marle, A.

AU - Hulscher, ST

AU - Selman, M.

AU - van Iperen, D.J.

AU - van Altena, K.

AU - Gillard, M.R.

AU - Bakker, R.A.

AU - Irth, H.

AU - Leurs, R.

AU - Vermeulen, N.P.E.

PY - 2007

Y1 - 2007

N2 - A flow-through fluorescence polarization (FP) detection system that makes use of a novel high-performance liquid chromatography (HPLC) fluorescence detector modified with polarization filters was developed. This flow-through FP detection system was evaluated by using a novel and very cost-effective bioassay for cyclic adenosine monophosphate (cAMP). The bioassay was first evaluated and optimized in an FP plate reader format and subsequently in a flow-through bioassay setup. The principle of the bioassay is based on the competition of cAMP and a fluorescent cAMP derivative for the cAMP binding domain of protein kinase A. cAMP could accurately be determined over a range of 0.8 to 30 pmol/well in the plate reader FP assay and over a range of 0.3 to 50 pmol/well in the flow-through FP assay setup. High Z′ factors (i.e., 0.89 for the plate reader and 0.93 for the flow-through FP cAMP assay, respectively) indicated robust assays. Finally, functional cAMP signaling of the human histamine H

AB - A flow-through fluorescence polarization (FP) detection system that makes use of a novel high-performance liquid chromatography (HPLC) fluorescence detector modified with polarization filters was developed. This flow-through FP detection system was evaluated by using a novel and very cost-effective bioassay for cyclic adenosine monophosphate (cAMP). The bioassay was first evaluated and optimized in an FP plate reader format and subsequently in a flow-through bioassay setup. The principle of the bioassay is based on the competition of cAMP and a fluorescent cAMP derivative for the cAMP binding domain of protein kinase A. cAMP could accurately be determined over a range of 0.8 to 30 pmol/well in the plate reader FP assay and over a range of 0.3 to 50 pmol/well in the flow-through FP assay setup. High Z′ factors (i.e., 0.89 for the plate reader and 0.93 for the flow-through FP cAMP assay, respectively) indicated robust assays. Finally, functional cAMP signaling of the human histamine H

U2 - 10.1177/1087057107308881

DO - 10.1177/1087057107308881

M3 - Article

SN - 1087-0571

VL - 12

SP - 1074

EP - 1083

JO - Journal of Biomolecular Screening

JF - Journal of Biomolecular Screening

IS - 8

ER -

A flow-through fluorescence polarization detection system for measuring GPCR-mediated modulation of cAMP production

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