title: Covalent organic framework-coated magnetic graphene as a novel support for trypsin immobilization.
abstract: Deep and efficient proteolysis is the critical premise in mass spectrometry-based bottom-up proteomics. It is difficult for traditional in-solution digestion to meet the requirement unless prolonged digestion time and enhanced enzyme dosage are employed, which makes the whole workflow time-consuming and costly. The abovementioned problems could be effectively ameliorated by anchoring many proteases on solid supports. In this work, covalent organic framework-coated magnetic graphene (MG@TpPa-1) was designed and prepared as a novel enzyme carrier for the covalent immobilization of trypsin with a high degree of loading (up to 268 μg mg-1). Profiting from the advantages of magnetic graphene and covalent organic frameworks, the novel trypsin bioreactor was successfully applied for the enzymatic digestion of a model protein with dramatically improved digestion efficiency, stability, and reusability. Complete digestion could be achieved in a time period as short as 2 min. For the digestion of proteins extracted from Amygdalus pedunculata, a total of 2833 protein groups were identified, which was slightly more than those obtained by 12 h of in-solution digestion (2739 protein groups). All of the results demonstrate that MG@TpPa-1-trypsin is an excellent candidate for sample preparation in a high-throughput proteomics analysis. Graphical abstract Covalent organic frameworks-coated magnetic graphene was prepared as novel carrier for highly efficient tryptic immobilization.
authors.affiliation: Key Laboratory of Synthetic and Natural Function Molecule Chemistry of Ministry of Education, College of Chemistry and Materials Science, Northwest University, Xi'an, Shaanxi, 710069, China.|State Key Laboratory of Proteomics, National Center for Protein Science Beijing, Beijing Institute of Radiation Medicine, Beijing, 102206, China.
authors.forename: Heping
authors.initials: H
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Proteomics, National Center for Protein Science Beijing, Beijing Institute of Radiation Medicine, Beijing, 102206, China.
authors.forename: Fenglong
authors.initials: F
authors.lastname: Jiao
authors.affiliation: State Key Laboratory of Proteomics, National Center for Protein Science Beijing, Beijing Institute of Radiation Medicine, Beijing, 102206, China.
authors.forename: Fangyuan
authors.initials: F
authors.lastname: Gao
authors.affiliation: State Key Laboratory of Proteomics, National Center for Protein Science Beijing, Beijing Institute of Radiation Medicine, Beijing, 102206, China.
authors.forename: Xinyuan
authors.initials: X
authors.lastname: Zhao
authors.affiliation: State Key Laboratory of Proteomics, National Center for Protein Science Beijing, Beijing Institute of Radiation Medicine, Beijing, 102206, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Key Laboratory of Synthetic and Natural Function Molecule Chemistry of Ministry of Education, College of Chemistry and Materials Science, Northwest University, Xi'an, Shaanxi, 710069, China. yhshen@nwu.edu.cn.
authors.forename: Yehua
authors.initials: Y
authors.lastname: Shen
authors.affiliation: State Key Laboratory of Proteomics, National Center for Protein Science Beijing, Beijing Institute of Radiation Medicine, Beijing, 102206, China. 13683167093@163.com.
authors.forename: Yangjun
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Proteomics, National Center for Protein Science Beijing, Beijing Institute of Radiation Medicine, Beijing, 102206, China. 13911734119@163.com.
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Qian
mesh_terms: D004800:Enzymes, Immobilized / Q000737:chemistry*; D006108:Graphite / Q000737:chemistry*; D008280:Magnetics; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission; D009930:Organic Chemicals / Q000737:chemistry*; D014357:Trypsin / Q000737:chemistry*
chemical_list: D004800:Enzymes, Immobilized; D009930:Organic Chemicals; D006108:Graphite; D014357:Trypsin
keywords: Covalent organic framework; Immobilized trypsin; Magnetic graphene