Proteomics analysis of tissue and biofluids has been used to identify biomarker molecules and signatures in the clinical environment. The proteins composition of biofluid, such as serum or plasma, spans ten orders of magnitude and making it difficult to detect in the presence of the most abundant proteins. Although the analysis of top 1000 proteins can satisfy patient classification, identifying specific protein biomarkers requires the consistent measurement of less abundant components of the serum. Targeted proteomics, a profound proteomic analysis method, will allow the prior acquired information to measure a specific subset of proteins in the sample. A new targeted proteomics technology, 4D-PRM, enables faster and more accurate targeted proteomics detection, bringing the possibility of targeted proteomics in large cohorts.
4D-PRM introduces a fourth dimension and ion trickle proteomic, with the advantages of high reproducibility and accurate quantification, breaking the conventional proteomics limitations. 4D-PRM utilizes Tims for the fourth-dimensional separation, which improves the selectivity and sensitivity of peptide ions compared to 3D-PRM. Moreover, 4D-PRM enables faster and more accurate targeted proteomic assays. Overall, the high multiplexing capacity and short chromatographic separation utilization make 4D-PRM particularly useful for clinical applications and biomarker validation.
4D-PRM mode (Antoine Lesur et al, Chem 2021)
Creative Proteomics has launched the 4D-PRM based on TimsTOF Pro to validate 100 differential proteins in 60 min with gradient separations. The 4D-PRM is over 2 times higher throughput than 3D-PRM. Due to the high MS/MS scan speed of TimsTOF Pro, 4D-PRM can theoretically target more than 12 target ions per 100 ms Tims ramp time and more than 100 target ions in 1 s duty cycle and enables 4D-PRM to quantify more proteins in the same amount of time.
The acquisition window of 4D-PRM includes three dimensions: chromatographic retention time, ion mobility, and m/z. On TimsTOF Pro, when retention time and m/z of the precursor ions are the same, separation can still be done through ion mobility, enhancing the specificity and accuracy of quantification.
In 4D-PRM mode, the fast scanning capability allows acquiring more data and generating more MS/MS spectra with higher specificity, which significantly improves detection sensitivity and quantitative reliability.
Verification: differential protein verification, mutation and modified site confirmation, etc.
Biomarkers: Biomarker screening, verification, absolute quantification, etc.
Using advanced high-resolution mass spectrometers Thermo Orbitrap, Creative Proteomics can provide you with cutting-edge protein 4D-PRM targeting protein validation services for clinical applications and biomarker validation.
Stepwise 4D-PRM workflow (Antoine Lesur et al. Expert Review of Proteomics 2021)
Tims TOF pro
Animal and clinical tissue specimens: 100 mg/sample
Serum, plasma: 200 μL/sample
Cells, microorganisms: 1×107 cells/sample
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4D Proteomics with Data-Independent Acquisition (DIA)
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×Specializing in proteomics, Creative Proteomics offers cutting-edge protein analysis services. Our distinctive approach revolves around harnessing the power of DIA technology, enabling us to deliver precise and comprehensive insights that drive advancements in research and industry.