Department of Pharmaceutical Biosciences, Uppsala University, Uppsala, Sweden.
Electrophoresis. 2010 Aug;31(16):2722-9. doi: 10.1002/elps.201000243.
Although protein biomarkers have a great potential as biomarkers for diagnosis of diseases, they are seldom used in hospitals. There are many reasons for this, for instance, the difficulties to (i) find a biomarker for which the concentration in body fluids clearly differs between patients and healthy subjects, (ii) attain purification of the biomarker close to 100%, which is required for production of conventional protein antibodies as well as artificial gel antibodies for selective capture of a biomarker, (iii) design a standard curve for rapid and accurate determination of the concentration of the biomarker in the body fluid because of adsorption of the biomarker onto vials, pipettes, etc., (iv) determine accurately the sample volume delivered by a pipette, (v) avoid polymerization of the biomarker upon storage and to decide whether it is in the form not only of monomers, but also of dimers, trimers, etc., in the native state, (vi) determine the degree of possible glycosylation and amidation of the biomarker and (vii) decide whether glycosylation and amidation positively or negatively affects the possibility to use the protein as a biomarker. In this article, we discuss in quantitative terms the difficulties (iii-vii) and how to overcome them, which also may help to overcome the difficulty (ii), which in turn minimizes difficulty (i).
尽管蛋白质生物标志物在疾病诊断方面具有很大的潜力，但它们很少在医院中使用。造成这种情况的原因有很多，例如：（i）难以找到一种生物标志物，其在患者和健康受试者的体液中的浓度有明显差异；（ii）需要接近 100%的生物标志物纯度，这对于生产常规蛋白质抗体以及用于选择性捕获生物标志物的人工凝胶抗体都是必需的；（iii）设计用于快速准确地确定生物标志物在体液中浓度的标准曲线，因为生物标志物会吸附到小瓶、移液管等上；（iv）准确确定移液管所输送的样品体积；（v）避免生物标志物在储存过程中的聚合，并确定其在天然状态下不仅以单体形式，而且以二聚体、三聚体等形式存在；（vi）确定生物标志物可能的糖基化和酰胺化程度；以及（vii）确定糖基化和酰胺化是否会积极或消极地影响将蛋白质用作生物标志物的可能性。在本文中，我们将定量讨论（iii-vii）的困难以及如何克服这些困难，这也可能有助于克服困难（ii），从而最大限度地减少困难（i）。
