College of Chemistry, Beijing Normal University, Beijing, P R China.
Proteomics. 2011 Sep;11(17):3510-7. doi: 10.1002/pmic.201000742. Epub 2011 Aug 4.
Various methods have been applied for serum ferritin detection, however, these methods still have some limitations. Over the last few years, quantum dots (QDs) have become very attractive for immunoassays because of their enormous potentials in ultrasensitive analysis. In this study, a Western blotting method combined with QDs-labeled avidin-biotin system for detecting human serum ferritin was described. Meanwhile, the traditional diaminobenzidine (DAB)-horseradish peroxidase (HRP) method had been compared with the method. The linearity of this QDs-based Western blotting method was from 0.27 to 1.1 ng, and the quantification limit was 0.27 ng, the sensitivity was up to pictogram values. Real serum samples such as hepatoma, thalassemia patient and normal individual sera were analyzed, the analysis results demonstrated that there was significant difference in the concentrations of ferritin between patients and normal individual serum. Furthermore, the recovery of ferritin from the serum samples of patients ranged from 98.15 to 119.67%, and the RSD (relative standard deviation) ranged from 8.73 to 11.61%, the repeatabilities were well within the acceptable range, which revealed that this method is a stable and reproducible method for detecting serum ferritin and have potential application prospect in clinical laboratory of serum ferritin detection.
已经应用了各种方法来检测血清铁蛋白，然而，这些方法仍然存在一些局限性。近年来，由于量子点（QDs）在超灵敏分析中具有巨大的潜力，因此它们在免疫分析中变得非常有吸引力。在这项研究中，描述了一种结合量子点标记的亲和素-生物素系统的 Western 印迹法来检测人血清铁蛋白。同时，将该方法与传统的二氨基联苯胺（DAB）-辣根过氧化物酶（HRP）方法进行了比较。该基于 QDs 的 Western 印迹法的线性范围为 0.27 至 1.1ng，定量限为 0.27ng，灵敏度高达皮克值。分析了肝癌、地中海贫血患者和正常个体血清等真实血清样本，分析结果表明，患者和正常个体血清中铁蛋白的浓度存在显著差异。此外，从患者血清样本中回收的铁蛋白回收率范围为 98.15%至 119.67%，相对标准偏差（RSD）范围为 8.73%至 11.61%，重复性均在可接受范围内，这表明该方法是一种稳定且可重复的检测血清铁蛋白的方法，在临床实验室检测血清铁蛋白方面具有潜在的应用前景。
