Citation: FAN Chao, SONG Zifeng, QIN Weijie, CAI Yun, QIAN Xiaohong. A novel immobilized trypsin on hydrophilic polymer modified silica beads for proteome characterization[J]. Chinese Journal of Chromatography, ;2013, 31(5): 423-428. doi: 10.3724/SP.J.1123.2013.01007 shu

A novel immobilized trypsin on hydrophilic polymer modified silica beads for proteome characterization

FAN Chao ^1,2 ,
SONG Zifeng ² ,
QIN Weijie ^2,, ,
CAI Yun ^2,, ,
QIAN Xiaohong ²

1.
1. Graduate Department of Anhui Medical University, Hefei 230032, China;

Corresponding author: QIN Weijie, CAI Yun,
Received Date: 5 January 2013
Available Online: 7 February 2013
Fund Project: 国家重大科学计划项目(2013CB911204) (2013CB911204)国家重大科学仪器设备开发专项项目(2011YQ09000504) (2011YQ09000504)国家自然科学基金项目(21275005,21235001). (21275005,21235001)

A new type of immobilized trypsin was prepared using hydrophilic polymer modified silica microparticles (HPMSM) synthesized by atom transfer radical polymerization (ATRP) as the substrate material. ATRP modification led to densely packed hydrophilic polymer chains grafted on the microparticles surface which resulted in largely increased trypsin loading amount and minimized the nonspecific adsorption of proteins and peptides. Therefore, ultra-fast and highly efficient protein digestion was achieved with minimized sample loss. For standard protein bovine serum albumin (BSA), 1 min digestion led to the identification of 93 peptides, which covered 91% amino acid sequence of the protein. This immobilized trypsin was further successfully applied to the digestion of complex protein samples from yeast and 666 proteins were identified after 3 min digestion, which exceeded the number of identified proteins after 12 h solution digestion.
- immobilized trypsin,
- proteomics,
- atom transfer radical polymerization (ATRP)
1. [1]
  [1] Elias J E, Gygi S P. Nat Methods, 2007, 4(3): 207
2. [2]
  [2] Elias J E, Haas W, Faherty B K, et al. Nat Methods, 2005, 2(9): 667
3. [3]
  [3] Ahrens C H, Brunner E, Qeli E, et al. Nat Rev Mol Cell Biol, 2010, 11(11): 789
4. [4]
  [4] Kocher T, Superti-Furga G. Nat Methods, 2007, 4(10): 807
5. [5]
  [5] Ren D, Pipes G D, Liu D J, et al. Anal Biochem, 2009, 392(1): 12
6. [6]
  [6] Lin S, Yao G P, Qi D W, et al. Anal Chem, 2008, 80(10): 3655
7. [7]
  [7] Qian K, Wan J J, Qiao L, et al. Anal Chem, 2009, 81(14): 5749
8. [8]
  [8] Li Y, Xu X Q, Deng C H, et al. J Proteome Res, 2007, 6(9): 3849
9. [9]
  [9] Liang Y, Tao D Y, Ma J F, et al. J Chromatogr A, 2011, 1218(20): 2898
10. [10]
  [10] Ma J F, Liang Z, Qiao X Q, et al. Anal Chem, 2008, 80(8): 2949
11. [11]
  [11] Ma J F, Liu J X, Sun L L. Anal Chem, 2009, 81(15): 6534
12. [12]
  [12] Calleri E, Temporini C, Gasparrini F, et al. J Chromatogr A, 2011, 1218(49): 8937
13. [13]
  [13] Jiang H, Yuan H M, Liang Y, et al. J Chromatogr A, 2012, 1246: 111
14. [14]
  [14] Lutz J F. Adv Mater, 2011, 23(19): 2237
15. [15]
  [15] Pokorski J K, Breitenkamp K, Liepold L O, et al. J Am Chem Soc, 2011, 133(24): 9242
16. [16]
  [16] Raynor J E, Petrie T A, Garcia A J, et al. Adv Mater, 2007, 19(13): 1724
17. [17]
  [17] Song Z F, Zhang Q L, Zhang Y J, et al. Chinese Journal of Chromatography (宋子凤, 张庆林, 张养军, 等. 色谱), 2012, 30(6): 549
18. [18]
  [18] Krogh T N, Berg T, Hojrup P. Anal Biochem, 1999, 274(2): 153
1. [1]
  Xinyi Hong , Tailing Xue , Zhou Xu , Enrong Xie , Mingkai Wu , Qingqing Wang , Lina Wu . Non-Site-Specific Fluorescent Labeling of Proteins as a Chemical Biology Experiment. University Chemistry, 2024, 39(4): 351-360. doi: 10.3866/PKU.DXHX202310010
2. [2]
  Xinran Zhang , Siqi Liu , Yichi Chen , Qingli Zou , Qinghong Xu , Yaqin Huang . From Protein to Energy Storage Materials: Edible Gelatin Jelly Electrolyte. University Chemistry, 2025, 40(7): 255-266. doi: 10.12461/PKU.DXHX202408104
3. [3]
  Danqing Wu , Jiajun Liu , Tianyu Li , Dazhen Xu , Zhiwei Miao . Research Progress on the Simultaneous Construction of C—O and C—X Bonds via 1,2-Difunctionalization of Olefins through Radical Pathways. University Chemistry, 2024, 39(11): 146-157. doi: 10.12461/PKU.DXHX202403087
4. [4]
  Jiajia Li , Xiangyu Zhang , Zhihan Yuan , Zhengyang Qian , Jian Zhu . 3D Printing Based on Photo-Induced Reversible Addition-Fragmentation Chain Transfer Polymerization. University Chemistry, 2024, 39(5): 11-19. doi: 10.3866/PKU.DXHX202309073
5. [5]
  Tongyan Yu , Pan Xu . Visible-Light Photocatalyzed Radical Rearrangement Reaction. University Chemistry, 2025, 40(7): 169-176. doi: 10.12461/PKU.DXHX202409070
6. [6]
  Zijian Zhao , Yanxin Shi , Shicheng Li , Wenhong Ruan , Fang Zhu , Jijun Jiang . A New Exploration of the Preparation of Polyacrylic Acid by Free Radical Polymerization Based on the Concept of Green Chemistry. University Chemistry, 2024, 39(5): 315-324. doi: 10.3866/PKU.DXHX202311094
7. [7]
  Zhongyan Cao , Shengnan Jin , Yuxia Wang , Yiyi Chen , Xianqiang Kong , Yuanqing Xu . Advances in Highly Selective Reactions Involving Phenol Derivatives as Aryl Radical Precursors. University Chemistry, 2025, 40(4): 245-252. doi: 10.12461/PKU.DXHX202405186
8. [8]
  Lei Shi . Nucleophilicity and Electrophilicity of Radicals. University Chemistry, 2024, 39(11): 131-135. doi: 10.3866/PKU.DXHX202402018
9. [9]
  Min LIU , Huapeng RUAN , Zhongtao FENG , Xue DONG , Haiyan CUI , Xinping WANG . Neutral boron-containing radical dimers. Chinese Journal of Inorganic Chemistry, 2025, 41(1): 123-130. doi: 10.11862/CJIC.20240362
10. [10]
  Baitong Wei , Jinxin Guo , Xigong Liu , Rongxiu Zhu , Lei Liu . Theoretical Study on the Structure, Stability of Hydrocarbon Free Radicals and Selectivity of Alkane Chlorination Reaction. University Chemistry, 2025, 40(3): 402-407. doi: 10.12461/PKU.DXHX202406003
11. [11]
  Dan Liu . 可见光-有机小分子协同催化的不对称自由基反应研究进展. University Chemistry, 2025, 40(6): 118-128. doi: 10.12461/PKU.DXHX202408101
12. [12]
  Xinxin Wu . 基础有机化学教学中自由基重排反应的课程设计及其课程思政元素的融入. University Chemistry, 2025, 40(6): 316-325. doi: 10.12461/PKU.DXHX202408055
13. [13]
  Heng Zhang . Determination of All Rate Constants in the Enzyme Catalyzed Reactions Based on Michaelis-Menten Mechanism. University Chemistry, 2024, 39(4): 395-400. doi: 10.3866/PKU.DXHX202310047
14. [14]
  Jinyao Du , Xingchao Zang , Ningning Xu , Yongjun Liu , Weisi Guo . Electrochemical Thiocyanation of 4-Bromoethylbenzene. University Chemistry, 2024, 39(6): 312-317. doi: 10.3866/PKU.DXHX202310039
15. [15]
  Chunmei GUO , Weihan YIN , Jingyi SHI , Jianhang ZHAO , Ying CHEN , Quli FAN . Facile construction and peroxidase-like activity of single-atom platinum nanozyme. Chinese Journal of Inorganic Chemistry, 2024, 40(9): 1633-1639. doi: 10.11862/CJIC.20240162
16. [16]
  .
  CCS Chemistry | 超分子活化底物为自由基促进高效选择性光催化氧化
  . CCS Chemistry, 2025, 7(10.31635/ccschem.025.202405229): -.
17. [17]
  Yinjie Xu , Suiqin Li , Lihao Liu , Jiahui He , Kai Li , Mengxin Wang , Shuying Zhao , Chun Li , Zhengbin Zhang , Xing Zhong , Jianguo Wang . Enhanced Electrocatalytic Oxidation of Sterols using the Synergistic Effect of NiFe-MOF and Aminoxyl Radicals. Acta Physico-Chimica Sinica, 2024, 40(3): 2305012-0. doi: 10.3866/PKU.WHXB202305012
18. [18]
  Yu Dai , Xueting Sun , Haoyu Wu , Naizhu Li , Guoe Cheng , Xiaojin Zhang , Fan Xia . Determination of the Michaelis Constant for Gold Nanozyme-Catalyzed Decomposition of Hydrogen Peroxide. University Chemistry, 2025, 40(5): 351-356. doi: 10.12461/PKU.DXHX202407052
19. [19]
  Wei HE , Jing XI , Tianpei HE , Na CHEN , Quan YUAN . Application of solar-driven inorganic semiconductor-microbe hybrids in carbon dioxide fixation and biomanufacturing. Chinese Journal of Inorganic Chemistry, 2025, 41(1): 35-44. doi: 10.11862/CJIC.20240364
20. [20]
  Yuan GAO , Yiming LIU , Chunhui WANG , Zhe HAN , Chaoyue FAN , Jie QIU . A hexanuclear cerium oxo cluster stabilized by furoate: Synthesis, structure, and remarkable ability to scavenge hydroxyl radicals. Chinese Journal of Inorganic Chemistry, 2025, 41(3): 491-498. doi: 10.11862/CJIC.20240271

Get Citation

PDF

XML

Metrics

PDF Downloads(0)
Abstract views(495)
HTML views(45)

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142