Departamento de Química, Facultad de Ciencias Exactas, Físico-Químicas y Naturales, Universidad Nacional de Río Cuarto, Agencia Postal N degrees 3, Río Cuarto, Argentina.
Talanta. 2010 Mar 15;80(5):1986-92. doi: 10.1016/j.talanta.2009.10.059. Epub 2009 Nov 1.
Progesterone (P4) is a steroidal hormone with a vital role in the maintenance of human and animal health. This paper describes the development of an immunosensor coupled to glassy carbon (GC) electrode and integrated to a microfluidic system to quantify P4 from bovine serum samples in a fast and sensitive way. The serum samples spiked with a given P4 concentration and a given P4 concentration bound to horseradish peroxide (HPR) were simultaneously added and, therefore, they competed immunologically with sheep monoclonal anti-P4 antibodies that were immobilized at a rotating disk. HRP in the presence of hydrogen peroxide (H(2)O(2)) catalyzes the chatecol (H(2)Q) oxidation to benzoquinone (Q). Its reverse electrochemical reduction to H(2)Q can be detected at a GC electrode surface at -0.15 V by chronoamperometric measurements. These current responses are proportional to the enzyme activity and inversely proportional to the P4 amount present in bovine serum samples. This P4 immunosensor showed a linear working range from 0.5 to 12.5 ng mL(-1). The detection (DL) and quantification (QL) limits were 0.2 and 0.5 ng mL(-1), respectively. The electrochemical immunosensor had a higher sensitivity than the ELISA method using conventional spectrophotometric detections. However, both methods allowed us to obtain similar detection limits. The immunosensor allowed us to make up to 100 determinations on different samples without any previous pre-treatment. This behavior proved to be suitable to detect P4 in routine veterinary, clinical, biological, physiological, and analytical assays.
孕酮（P4）是一种甾体激素，在维护人类和动物健康方面起着至关重要的作用。本文描述了一种免疫传感器的开发，该传感器与玻碳（GC）电极耦合，并集成到微流控系统中，以快速灵敏地定量牛血清样品中的 P4。用给定浓度的 P4 和与辣根过氧化物酶（HPR）结合的给定浓度的 P4 对血清样品进行了加标，然后同时加入，因此它们与固定在旋转盘上的绵羊单克隆抗 P4 抗体进行免疫竞争。在存在过氧化氢（H2O2）的情况下，辣根过氧化物酶（HRP）催化儿茶酚（H2Q）氧化为苯醌（Q）。其在 GC 电极表面的反向电化学还原可以通过计时安培测量法检测到 -0.15 V。这些电流响应与酶活性成正比，与牛血清样品中存在的 P4 量成反比。这种 P4 免疫传感器的工作线性范围从 0.5 到 12.5ng mL(-1)。检测（DL）和定量（QL）限分别为 0.2 和 0.5ng mL(-1)。电化学免疫传感器比使用传统分光光度检测的 ELISA 方法具有更高的灵敏度。然而，这两种方法都允许我们获得相似的检测限。该免疫传感器允许我们在无需任何预处理的情况下对不同的样品进行多达 100 次测定。这种行为被证明适用于常规兽医、临床、生物、生理和分析测定中 P4 的检测。
