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Discovery-based Quantitative MS methods

Discovery-based Quantitative MS methods,(“shotgun” proteomics), aim to identify as many proteins in a sample as possible and can be completed on a wide range of samples with minimal method development.

  • DDA (data-dependent acquisition): In Data-dependent acquisition (DDA) mode a MS instrument will complete a full scan of all peptide (precursor) ions during MS1 and then select the most abundant ions for fragmentation and MS2
  • DIA (data-independent acquisition): In Data-independent acquisition (DIA) mode a MS instrument scans pre-determined, often sequential, m/z ranges and then fragments all the ions within each range for MS2 measurement

DDA vs DIA MS1 and MS2 examples

  DDA DIA
Suitable for small proteomes* Yes Yes
Suitable for large proteomes* No Yes
Dynamic range Moderate High
Complexity of spectra Low High
Search requirements Protein database Spectral library
Instrument method optimization Low Hig

Note (*) Most prokaryote proteomes can be considered a “small” proteome, vs. larger eukaryotic or mixed (e.g., infection model) proteomes

Discovery-based methods can be used to identify thousands of proteins in a single sample, and for most research questions label-free DDA is a straight-forward option that has relatively low reagents costs, and requires minimal sample preparation and instrument method optimization.  

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Source URL:https://www.uoguelph.ca/aac/facilities/mass-spectrometry-facility/quantitative-proteomics/discovery-based-quantitative-ms