Proteome-Wide Multipoint Internal Calibration Curves for Evaluating Peptide-Level Linearity in Relative Quantitative...

Area of Science:

• Proteomics
• Mass Spectrometry
• Cancer Research

Background:

• Mass spectrometry (MS)-based proteomics offers accurate peptide and protein quantification via calibration curves.
• External calibration curves can be affected by sample matrix effects.
• Internal calibration curves address matrix effects but present scalability and cost issues for proteome-wide analyses.

Purpose of the Study:

• To develop a novel TMT-based multipoint internal calibration curve strategy for proteome-wide linearity assessment.
• To evaluate the quantitative linearity of all identified peptides within a single experiment.
• To investigate proteome alterations in human ovarian cancer cells following cisplatin treatment.

Main Methods:

• Implementation of a TMT-based multipoint internal calibration curve strategy.
• Application of the strategy to human ovarian cancer cell samples.
• Analysis of peptide linearity and proteome changes post-cisplatin treatment.

Main Results:

• Successful generation of internal calibration curves for all identified peptides.
• Demonstration of linear quantitative responses across the proteome.
• Identification of significant proteome changes induced by cisplatin treatment in ovarian cancer cells.

Conclusions:

• The novel TMT-based multipoint internal calibration strategy enables robust proteome-wide quantitative linearity assessment.
• This method facilitates the accurate analysis of proteome dynamics in response to therapeutic agents like cisplatin.
• The findings contribute to a deeper understanding of ovarian cancer biology and potential therapeutic targets.