Liu Bing, Tang Hanyu, Liu Qian, Wang Wenwen, Li Haitao, Zheng Shiya, Sun Fei, Zhao Xiangwei
Medical School, Institute of Reproductive Medicine, Nantong University, Nantong, 226001, China.
Medical School, Institute of Reproductive Medicine, Nantong University, Nantong, 226001, China.
Talanta. 2023 Feb 1;253:123888. doi: 10.1016/j.talanta.2022.123888. Epub 2022 Sep 3.
Western blot (WB) is the most commonly used scheme for protein identification in life science, but it still faces great challenges in the accurate quantitative detection of low-abundance proteins. Here, we proposed a novel surface-enhanced Raman scattering-based Western blot (SERS-WB) to solve this challenge. SERS nanotags were used as quantitative labels of proteins, which were composed of gold-silver core-shell nanoparticles, and Nile blue A (NBA) molecules were anchored on the interface of the core and shell. The results show that the SERS-WB possessed excellent sensitivity with detection limit of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein of 0.15 pg, as well as wide linear dynamic range (LDR) of 382 fg to 382 ng. In addition, the target protein on nitrocellulose (NC) membrane could be directly identified by colorimetric signal due to the aggregation effect of nanoparticles, which greatly simplifies the procedure. This as-proposed strategy will bring new thoughts to technological innovation of WB.
蛋白质免疫印迹法(WB)是生命科学中最常用的蛋白质鉴定方法,但在低丰度蛋白质的准确定量检测方面仍面临巨大挑战。在此,我们提出了一种基于表面增强拉曼散射的新型蛋白质免疫印迹法(SERS-WB)来解决这一挑战。SERS纳米标签用作蛋白质的定量标记,由金银核壳纳米颗粒组成,尼罗蓝A(NBA)分子锚定在核壳界面上。结果表明,SERS-WB具有出色的灵敏度,甘油醛-3-磷酸脱氢酶(GAPDH)蛋白的检测限为0.15 pg,线性动态范围(LDR)宽,为382 fg至382 ng。此外,由于纳米颗粒的聚集效应,硝酸纤维素(NC)膜上的目标蛋白可以通过比色信号直接鉴定,这大大简化了操作过程。该策略将为WB的技术创新带来新的思路。