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Label-free DIA Quantitation

Service Description

BGI participates in quantitative performance assessment as well as standardization and harmonization of Multi-National DIA proteomics analysis supporting precision medicine studies. We have developed innovative solutions for optimizing the process of accurately quantifying proteins in complex biological matrices and other complex mixtures.

 

Label-Free DIA Quantitative Proteomics

Data-Independent Acquisition (DIA) is a label-free quantitative technique which provides highly consistent quantitation and broad proteome coverage¹. We create a customized spectral library with your specific sample and then analyze individual samples using nano-flow LC-MS/MS with DIA scanning and quantitation². Label-Free DIA Quantitative Proteomics service is ideal for long-term projects or projects with large sample sets which require accurate and reproducible quantitation.

 

Project Workflow

  • Sample preparation
  • HPLC-UV Fractionation
  • LC-MS/MS DDA Analysis
  • DDA library generation and updates
  • LC-MS/MS DIA analysis
  • Quantitation and bioinformatics

 

Mass Spectrometry Service Specification

Label-Free DIA Quantitative Proteomics services are performed using nano-flow liquid chromatography and cutting-edge mass spectrometer (Q Exactive HF/HF-X and Orbitrap Fusion Lumos).

 

Sample Preparation and Services

  • The QC analysis of enzymatic hydrolysis is strictly controlled by LC-MS
  • DDA-Spectral Library generated using HPLC technology and sample fractionation
  • 2h/fraction for non blood; and 2h/fraction for non-human blood; 1h/fraction for human blood

 

Quality Standard

  • Summary includes all methods and data analysis
  • Reports provided in Excel or PDF format, RAW files available upon request

 

Turn Around Time
Typical 4-5 weeks from sample QC acceptance to data report delivery

 

Sample Requirements

We accept protein samples in a variety of formats. For attaining maximum proteome coverage we recommend utilizing our sample fractionation services, performed using our off-line UHPLC-UV platform.

Sample type

Amount and Concentration

Minimum sample volume

Protein sample in liquid solution

Recommended 400 µg with fractionation: 2 µg/µL

200 µL

Protein sample in liquid solution

Minimum Required 100 µg with fractionation: 1 µg/µL

100 µL

 

Data Analysis

ADVANCED BIOINFORMATICS

  • Data analysis and validation performed with Mascot
  • Protein identification and quantification list
  • Differential proteins data statistics and volcano plot
  • Principal component analysis (PCA) and cluster analysis
  • Protein GO/COG/KOG/Pathway annotation
  • GO/COG/KOG enrichment analysis of differential proteins
  • Protein-protein interaction analysis
  • Protein subcellular localization analysis
  • Time series analysis
  • Repeatability analysis
  • Quantitative proteomics and phosphoproteomics correlation analysis