Quantitative Comparisons of Large Numbers of Human Plasma Samples Using TMT10plex Labeling

Area of Science:

• Proteomics
• Biomarker Discovery
• Mass Spectrometry

Background:

• Improving throughput in human plasma proteomic discovery is crucial for identifying biomarkers.
• Multiplexing strategies using isobaric tags, such as the TMT10plex kit, offer enhanced analytical capacity.

Purpose of the Study:

• To describe an efficient shotgun discovery proteomics strategy for analyzing large cohorts of human plasma.
• To leverage the TMT10plex kit for high-throughput proteomic analysis and biomarker identification.

Main Methods:

• Utilized isobaric tags (TMT10plex) for multiplexing human plasma samples.
• Employed a common pooled reference strategy for cross-experiment comparisons.
• Applied data filters, including precursor ion purity and unique peptide selection, to enhance quantitation quality.

Main Results:

• Demonstrated excellent overall reproducibility across different TMT experiments using duplicate samples.
• Successfully identified potential biomarkers through efficient shotgun discovery proteomics of large plasma cohorts.
• Validated the effectiveness of data filters in improving peptide and protein quantitation quality.

Conclusions:

• Multiplexing with TMT10plex kits is an effective strategy for high-throughput human plasma proteomic discovery.
• The described analysis strategy, incorporating pooled references and rigorous data filtering, enables robust biomarker identification.