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Open Access Research

In vitro detection of adrenocorticotropic hormone levels by fluorescence correlation spectroscopy immunoassay for mathematical modeling of glucocorticoid-mediated feedback mechanisms

Martin Gerald Puchinger1*, Clemens Alexander Zarzer2, Philipp Kügler2, Erwin Gaubitzer1 and Gottfried Köhler1

Author Affiliations

1 Department of Structural and Computational Biology, Max F. Perutz Laboratories (MFPL), University of Vienna, Campus-Vienna-Biocenter 5, Vienna, 1030, Austria

2 Johann Radon Institute for Computational and Applied Mathematics (RICAM), Austrian Academy of Sciences, Altenbergerstr. 69, Linz, 4040, Austria

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EURASIP Journal on Bioinformatics and Systems Biology 2012, 2012:17  doi:10.1186/1687-4153-2012-17

Published: 26 October 2012

Abstract

Performing quantitative, highly sensitive measurements at a single molecule level is often necessary to address specific issues related to complex molecular and biochemical systems. For that purpose, we present a technique exploiting both the flexibility of immunoassays as well as the low operating costs and high throughput rates of the fluorescence correlation spectroscopy (FCS) method. That way we have established a quantitative measurement technique providing accurate and flexibly time resolved data of single molecules. Nanomolar changes in adrenocorticotropic hormone (ACTH) levels have been detected in a short time-frame that are caused by fast feedback actions in AtT-20 anterior pituitary glands in vitro. Especially with respect to clinical diagnostic or mathematical modeling this improved FCS setup may be of high relevance in order to accurately quantify the amounts of peptide hormones—such as ACTH—as well as signaling molecules, transcription factors, etc., being involved in intra- and extracellular reaction networks.

Keywords:
ACTH; FCS; AtT-20; Cortisol; CRH; Glucocorticoid membrane receptor; ODE model; Parameter identification