High-throughput proteomics.

  title={High-throughput proteomics.},
  author={Zhaorui Zhang and Si Wu and David L Stenoien and Ljiljana Pa{\vs}a-Toli{\'c}},
  journal={Annual review of analytical chemistry},
Mass spectrometry (MS)-based high-throughput proteomics is the core technique for large-scale protein characterization. Due to the extreme complexity of proteomes, sophisticated separation techniques and advanced MS instrumentation have been developed to extend coverage and enhance dynamic range and sensitivity. In this review, we discuss the separation and prefractionation techniques applied for large-scale analysis in both bottom-up (i.e., peptide-level) and top-down (i.e., protein-level… 

High-throughput Proteomics Enabled by a Photocleavable Surfactant.

A high-throughput bottom-up proteomics method enabled by a newly developed MS-compatible photocleavable surfactant, 4-hexylphenylazosulfonate (Azo) that facilitates robust protein extraction, rapid enzymatic digestion, and subsequent MS-analysis following UV degradation is reported for the first time.

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

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 is developed.

Enhancing Sensitivity of Microflow-Based Bottom-Up Proteomics through Postcolumn Solvent Addition.

The presented microflow-LC-MS workflow, including postcolumn solvent addition, can be easily adopted on any LC-MS/MS platform and resulted in an overall 28-36% increase in identifications at both the protein and peptide levels.

MASH Suite Pro: A Comprehensive Software Tool for Top-Down Proteomics*

MASH Suite Pro significantly simplifies and speeds up the interpretation of high-resolution top-down proteomics data by integrating tools for protein identification, quantitation, characterization, and visual validation into a customizable and user-friendly interface.

Chromatographic separation strategies for precision mass spectrometry to study protein-protein interactions and protein phosphorylation.

  • Z. MinićT. DahmsM. Babu
  • Biology, Chemistry
    Journal of chromatography. B, Analytical technologies in the biomedical and life sciences
  • 2018

Two-Dimensional Separation Using High-pH and Low-pH Reversed Phase Liquid Chromatography for Top-down Proteomics.

Use of monolithic supports for high‐throughput protein and peptide separation in proteomics

The chromatographic conditions and the mass spectrometric data acquisition conditions are discussed and an attempt is made to present currently demonstrated capacities of monolithic capillary columns for HTP identification and quantification of proteins and peptides from complex biological samples by MS‐based proteomics.

Identification and Quantification of Proteoforms by Mass Spectrometry

Recent technological developments have enabled comprehensive proteoform analyses in complex samples, and an increasing number of laboratories are adopting top‐down proteomic workflows.

Identification of differentially expressed peptides in high‐throughput proteomics data

It is found that three or four replicates are the minimum requirement for high‐throughput data analysis and confident assignment of significant changes and empirical Bayes method (limma) achieves the highest sensitivity, and is recommended for performing differential expression analysis at the peptide level.