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

Martin Gerald Puchinger, Clemens Alexander Zarzer, Philipp Kügler, Erwin Gaubitzer, Gottfried Köhler
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引用次数: 5

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.

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用荧光相关光谱免疫分析法体外检测促肾上腺皮质激素水平,建立糖皮质激素介导反馈机制的数学模型。
在单分子水平上进行定量,高灵敏度的测量通常是解决与复杂分子和生化系统相关的特定问题所必需的。为此,我们提出了一种利用免疫测定的灵活性以及荧光相关光谱(FCS)方法的低操作成本和高通量率的技术。通过这种方式,我们建立了一种定量测量技术,提供了准确灵活的单分子时间分辨数据。促肾上腺皮质激素(ACTH)水平的纳摩尔变化已在短时间内被检测到,这是由体外at -20垂体前腺的快速反馈作用引起的。特别是在临床诊断或数学建模方面,这种改进的FCS设置可能与准确量化参与细胞内和细胞外反应网络的肽激素(如acth)以及信号分子、转录因子等的数量高度相关。
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