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用于临床样本中生物标志物检测和定量的高选择性人工凝胶抗体。I. 用于设计定量校准曲线的分光光度法。

Highly selective artificial gel antibodies for detection and quantification of biomarkers in clinical samples. I. Spectrophotometric approach to design the calibration curve for the quantification.

作者信息

Ghasemzadeh Nasim, Nyberg Fred, Hjertén Stellan

机构信息

Department of Pharmaceutical Biosciences, Uppsala University, Uppsala, Sweden.

出版信息

J Sep Sci. 2008 Dec;31(22):3945-53. doi: 10.1002/jssc.200800385.

Abstract

High selectivity of a biomarker is a basic requirement when it is used for diagnosis, prognosis and treatment of a disease. The artificial gel antibodies, which we synthesise by a molecular imprinting method, have this property not only for proteins, but also for bioparticles, such as viruses and bacteria. However, diagnosis of a disease requires not only that the biomarker can be "fished out" from a body fluid with high selectivity, but also that its concentration in the sample can rapidly be determined and preferably by a simple technique. This paper deals primarily with the development of a spectrophotometric method, which is so simple and fast that it can be used with advantage in a Doctor's Office. The development of this method was not straight-forward. However, by modifications of the performance of these measurements we can now design standard curves in the form of a straight line, when we plot the true (not the recorded "apparent" absorption) against known protein concentrations. In an additional publication (see the following paper in this issue of JSS) we show an application of such a plot: determination of the concentration of albumin in serum and cerebrospinal fluid from patients with neurological disorders to investigate whether albumin is a biomarker for these diseases.

摘要

生物标志物的高选择性是其用于疾病诊断、预后和治疗的基本要求。我们通过分子印迹法合成的人工凝胶抗体不仅对蛋白质具有这种特性,对生物颗粒(如病毒和细菌)也具有此特性。然而,疾病诊断不仅要求生物标志物能够以高选择性从体液中“钓出”,还要求能够快速测定其在样品中的浓度,并且最好采用简单的技术。本文主要探讨一种分光光度法的开发,该方法非常简单快速,可在医生办公室中很好地使用。该方法的开发并非一帆风顺。然而,通过对这些测量性能的改进,当我们将真实值(而非记录的“表观”吸光度)与已知蛋白质浓度作图时,现在我们可以设计出直线形式的标准曲线。在另一篇出版物中(见本期《日本光谱学会志》的后续文章),我们展示了这种作图方法的一个应用:测定神经疾病患者血清和脑脊液中的白蛋白浓度,以研究白蛋白是否为这些疾病的生物标志物。

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