Hollow silica bubble based immobilized trypsin for highly efficient proteome digestion and buoyant separation

RSC Advances ◽  
2016 ◽  
Vol 6 (87) ◽  
pp. 84113-84118 ◽  
Author(s):  
Fenglong Jiao ◽  
Rui Zhai ◽  
Junjie Huang ◽  
Yukui Zhang ◽  
Yangjun Zhang ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Tryptic Digestion ◽  
Hollow Silica ◽  
Highly Efficient ◽  
Immobilized Trypsin ◽  
Identification And Quantification

Tryptic digestion before identification and quantification by mass spectrometry is an indispensable process for most proteomics studies.

scholarly journals Ultrafast and Reproducible Proteomics from Small Amounts of Heart Tissue Enabled by Azo and timsTOF Pro

2021 ◽  
Author(s):  
Timothy J Aballo ◽  
David S Roberts ◽  
Jake A Melby ◽  
Kevin M Buck ◽  
Kyle A Brown ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Large Scale ◽  
Protein Identification ◽  
Heart Tissue ◽  
Tryptic Digestion ◽  
Bottom Up ◽  
Preparation Methods ◽  
Liquid Chromatography Tandem Mass ◽  
And Function ◽  
Identification And Quantification

Global bottom-up mass spectrometry (MS)-based proteomics is widely used for protein identification and quantification to achieve a comprehensive understanding of the composition, structure, and function of the proteome. However, traditional sample preparation methods are time-consuming, typically including overnight tryptic digestion, extensive sample clean-up to remove MS-incompatible surfactants, and offline sample fractionation to reduce proteome complexity prior to online liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Thus, there is a need for a fast, robust, and reproducible method for protein identification and quantification from complex proteomes. Herein, we developed an ultrafast bottom-up proteomics method enabled by Azo, a photocleavable, MS-compatible surfactant that effectively solubilizes proteins and promotes rapid tryptic digestion, combined with the Bruker timsTOF Pro, which enables deeper proteome coverage through trapped ion mobility spectrometry (TIMS) and parallel accumulation-serial fragmentation (PASEF) of peptides. We applied this method to analyze the complex human cardiac proteome and identified nearly 4,000 protein groups from as little as 1 mg of human heart tissue in a single one-dimensional LC-TIMS-MS/MS run with high reproducibility. Overall, we anticipate this ultrafast, robust, and reproducible bottom-up method empowered by both Azo and the timsTOF Pro will be generally applicable and greatly accelerate the throughput of large-scale quantitative proteomic studies. Raw data are available via the MassIVE repository with identifier MSV000087476.

scholarly journals Multiple-Approaches to the Identification and Quantification of Cytochromes P450 in Human Liver Tissue by Mass Spectrometry

2009 ◽  
Vol 8 (4) ◽  
pp. 1672-1681 ◽  
Author(s):  
Cathrin Seibert ◽  
Brian R. Davidson ◽  
Barry J. Fuller ◽  
Laurence H. Patterson ◽  
William J. Griffiths ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Liver Tissue ◽  
Human Liver ◽  
Cytochromes P450 ◽  
Human Liver Tissue ◽  
Identification And Quantification
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