Immobilization of trypsin onto multifunctional meso-/macroporous core-shell microspheres: a new platform for rapid enzymatic digestion.
A simple, fast, efficient, and reusable microwave-assisted tryptic digestion system which was constructed by immobilization of trypsin onto porous core-shell Fe3O4@fTiO2 microspheres has been developed. The nanostructure with magnetic core and titania shell has multiple pore sizes (2.4 and 15.0 nm), high pore volume (0.25 cm(3) g(-1)), and large surface area (50.45 m(2) g(-1)). For the proteins, the system can realize fast and efficient microwave-assisted tryptic digestion. Various standard proteins (e.g., cytochrome c (cyt-c), myoglobin (MYO), β-lactoglobulin (β-LG), and bovine serum albumin (BSA)) used can be digested in 45 s under microwave radiation, and they can be confidently identified by mass spectrometry (MS) analysis; even the concentration of substrate is as low as 5 ng μL(-1). Furthermore, the system for the 45 s microwave-assisted tryptic digestion is still effective after the trypsin-immobilized microspheres have been reused for 5 times. Importantly, 1715 unique proteins from 10 μg mouse brain proteins can be identified with high confidence after treatment of 45 s microwave-assisted tryptic digestion.
State Key Laboratory of Rare Earth Resource Utilization, Changchun Institute of Applied Chemistry, Changchun 130022, PR China; University of Chinese Academy of Sciences, Beijing 100049, PR China.
Gong
G
Cheng
Key Laboratory of Marine Bioresources and Ecology, College of Life Science, Shenzhen University, Shenzhen 518060, PR China.
Ping
P
Chen
State Key Laboratory of Rare Earth Resource Utilization, Changchun Institute of Applied Chemistry, Changchun 130022, PR China; University of Chinese Academy of Sciences, Beijing 100049, PR China.
Zhi-Gang
ZG
Wang
State Key Laboratory of Rare Earth Resource Utilization, Changchun Institute of Applied Chemistry, Changchun 130022, PR China; University of Chinese Academy of Sciences, Beijing 100049, PR China.
Xiao-Jing
XJ
Sui
State Key Laboratory of Rare Earth Resource Utilization, Changchun Institute of Applied Chemistry, Changchun 130022, PR China. Electronic address: zjl@ciac.ac.cn.
Ji-Lin
JL
Zhang
State Key Laboratory of Rare Earth Resource Utilization, Changchun Institute of Applied Chemistry, Changchun 130022, PR China; Key Laboratory of Marine Bioresources and Ecology, College of Life Science, Shenzhen University, Shenzhen 518060, PR China.
Jia-Zuan
JZ
Ni
D005290:Ferric Compounds / Q000737:chemistry; D013058:Mass Spectrometry; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission; D008863:Microspheres; D008872:Microwaves; D011506:Proteins / Q000737:chemistry; D013052:Spectrometry, X-Ray Emission; D014025:Titanium / Q000737:chemistry; D014357:Trypsin / Q000737:chemistry*; D014961:X-Ray Diffraction
D005290:Ferric Compounds; D011506:Proteins; C009495:titanium dioxide; C000499:ferric oxide; D014025:Titanium; D014357:Trypsin
Digestion; Magnetite; Mass spectrometry; Mesoporous; Titania; Trypsin