期刊
ANALYTICAL METHODS
卷 6, 期 8, 页码 2518-2525出版社
ROYAL SOC CHEMISTRY
DOI: 10.1039/c3ay41829d
关键词
-
资金
- National Key Program for Basic Research of China [2013CB911200, 2011CB910603]
- National Key Scientific Instrument Development Program of China [2011YQ09000504]
- National High-Tech Research and Development Program [2012AA020202]
- International Scientific Cooperation Project of China [2011DFB30370]
- National Natural Science Foundation of China [21275005, 21235001]
Protein N-glycosylation plays an important role in a variety of biological processes, and unusual changes in glycan structures are often correlated with various pathological states. Therefore, N-glycan profiling not only plays a vital role in the mechanism study of biological processes but also contributes to the discovery of biomarkers for early disease diagnosis. However, the conventional in-solution release of N-glycans by free endoglycosidases, such as peptide N-glycosidase F (PNGase F), suffers from several drawbacks, including prolonged incubation time, possible incomplete digestion and high cost due to the non-reusability of the enzyme. Herein, we report a novel graphene oxide (GO)-based immobilized PNGase F reagent for fast and highly efficient N-glycan release and glycoform profiling of glycoproteins. To create this reagent, PNGase F was attached to the GO surface via carbodiimide-activated amidation. The ultra-high surface area of GO allows for high loading with PNGase F and significantly facilitates the deglycosylation process. Highly efficient N-glycan release of ribonuclease B and asialofetuin was achieved with this reagent within 2 min. Furthermore, the successful application of our GO-based immobilized PNGase F in the N-glycan release of the human plasma glycoproteome was demonstrated by reproducible N-glycan identification using MALDI-TOF MS.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据