Niu Ji-Cheng, Zhou Ting, Niu Li-Li, Xie Zhen-Sheng, Fang Fang, Yang Fu-Quan, Wu Zhi-Yong
Research Center for Analytical Sciences, Chemistry Department College of Sciences, Northeastern University, Shenyang, Liaoning, 110819, China.
The Key Laboratory of Protein and Peptide Pharmaceuticals & Laboratory of Proteomics, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China.
Anal Bioanal Chem. 2018 Feb;410(6):1689-1695. doi: 10.1007/s00216-017-0809-5. Epub 2018 Jan 11.
In this work, fast isoelectric focusing (IEF) was successfully implemented on an open paper fluidic channel for simultaneous concentration and separation of proteins from complex matrix. With this simple device, IEF can be finished in 10 min with a resolution of 0.03 pH units and concentration factor of 10, as estimated by color model proteins by smartphone-based colorimetric detection. Fast detection of albumin from human serum and glycated hemoglobin (HBA1c) from blood cell was demonstrated. In addition, off-line identification of the model proteins from the IEF fractions with matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was also shown. This PAD IEF is potentially useful either for point of care test (POCT) or biomarker analysis as a cost-effective sample pretreatment method.
在本研究中,快速等电聚焦(IEF)在开放纸质流体通道上成功实现,用于从复杂基质中同时浓缩和分离蛋白质。利用这种简单的装置,通过基于智能手机的比色检测对颜色模型蛋白进行估算,IEF可在10分钟内完成,分辨率为0.03个pH单位,浓缩系数为10。已证明可快速检测人血清中的白蛋白和血细胞中的糖化血红蛋白(HBA1c)。此外,还展示了使用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)对IEF组分中的模型蛋白进行离线鉴定。这种纸质等电聚焦作为一种经济高效的样品预处理方法,在即时检验(POCT)或生物标志物分析中具有潜在用途。