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Home > Services > Proteomics Service > Protein Quantification > Parallel Reaction Monitoring (PRM)

Parallel Reaction Monitoring (PRM)

Parallel reaction monitoring (PRM) is an ion monitoring technique based on high-resolution and high-precision mass spectrometry. The principle of this technique is comparable to SRM/MRM, but it is more convenient in assay development for absolute quantification of proteins and peptides. It is most suitable for quantification of multiple proteins in complex sample with an attomole-level detection.

Parallel Reaction Monitoring (PRM)

PRM is based on Q-Orbitrap as the representative quadrupole-high resolution mass spectrum platform. Unlike the SRM, which performs one transition at a time, the PRM performs a full scan of each transition by a precursor ion, that is, parallel monitoring of all fragments from the precursor ion. First, the PRM uses the quadrupole (Q1) to select the precursor ion, and the selection window is usually m/z≤2; then, the precursor ion is fragmented in the collision cell (Q2); finally, Orbitrap replaces Q3, scans all product ions with high resolution and high accuracy. Therefore, PRM technology not only has the SRM/MRM target quantitative analysis capabilities, but also have the qualitative ability. (1) The mass accuracy can reach to ppm level, which can eliminate the background interference and false positive better than SRM / MRM, and improve the detection limit and sensitivity in complex background effectively; (2) Full scan of product ions, without the need to select the ion pair and optimize the fragmentation energy, easier to establish the assay; (3) a wider linear range: increased to 5-6 orders of magnitude.

Highlight of SRM/MRM technique:

  • High sensitivity;
  • High throughput;
  • Easier to develop assay.

PRM Analysis Services at Creative-Proteomics offers you a state-of-the-art strategy of proteins analysis. The workflow is based on the following sections:

  • The PRM assay development. This is the core of the approach. First, 2 or 3 targeted peptides should be selected from a spectral library generation or Skyline software. The selected peptides should be unique to the protein of interest and easily detected by LC-MS. They also require no missed cleavage sites and no frequently modified amino acids. Second, a working linear curve was built with the corresponding concentration ratios on the x-axis and peak area ratios on the y-axis.
  • Analysis of the sample. After digestion, the sample is performed on the Q-Oribtrap mass spectrometer, such as Q Exactive or Fusion mass spectrometer.
  • Data analysis. Data will be analyzed with Skyline software.

PRM application

  • Verification of iTRAQ differential protein;
  • Verification of label-free differential protein follow-up;
  • Protein and peptide absolute quantification;
  • Quantification of disease markers, the establishment of diagnostic models;
  • Phosphorylation protein quantification, methylation protein quantification;
  • Other post-translationally modified protein quantification;
  • Quantitative analysis of pathways.

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