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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pages = "1074--83",

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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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