Immobilization of trypsin on graphene oxide for microwave-assisted on-plate proteolysis combined with MALDI-MS analysis

The Analyst
Guobin XuLiming Wei

Abstract

With an ultra-high surface area and abundant functional groups, graphene oxide (GO) provides an ideal substrate for the immobilization of trypsin. We demonstrated that trypsin could be immobilized on GO sheets assisted by polymers as molecular spacers to maintain the activity of the enzyme. And with the trypsin-linked GO as the enzyme immobilization probe, a novel microwave-assisted on-plate digestion method has been developed with subsequent analysis by MALDI-MS. The feasibility and performance of the digestion approach were demonstrated by the proteolysis of standard proteins. The results show that this novel approach substantially accelerated proteolysis and reduced the time required for traditional procedures involving on-plate enzymatic digestion and sample preparation prior to MALDI-MS analysis. The novel digestion approach is simple and efficient, offering great promise for high throughput protein identification.

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Citations

Sep 2, 2014·Materials Science & Engineering. C, Materials for Biological Applications·Dolly GogoiJoyanti Chutia
Dec 27, 2015·International Journal of Biological Macromolecules·Michaela PatilaHaralambos Stamatis
Jan 13, 2016·Analytical and Bioanalytical Chemistry·Jing WangGuibin Jiang
Sep 12, 2015·Bioanalysis·Suresh Kumar KailasaHui-Fen Wu
Feb 18, 2014·Biochemical and Biophysical Research Communications·Zhengyi ChenGuihua Ruan
Jun 20, 2013·Journal of Pharmaceutical and Biomedical Analysis·Youji ShimazakiMasaki Saito
Dec 24, 2013·Journal of Chromatography. a·Muhammad SafdarJanne Jänis
May 6, 2014·Trends in Biotechnology·Ioannis V PavlidisHaralambos Stamatis
Jun 14, 2017·The Analyst·Jinlan DongMerlin L Bruening
Jun 4, 2019·Biosensors·Aditya BalajiJin Zhang
Dec 8, 2019·Mikrochimica Acta·Shubhi JoshiAvneet Saini
Aug 5, 2020·Biosensors & Bioelectronics·Eleni VermisoglouMichal Otyepka
Jan 27, 2018·International Journal of Biological Macromolecules·Faranak JafarianAhmadreza Khosropour

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