Two-dimensional SDS-PAGE fractionation of biological samples for biomarker discovery
Thierry Rabilloud (LCBM), Sarah Triboulet (LCBM)
TL;DR
This paper presents a comprehensive workflow for applying two-dimensional electrophoresis to analyze simpler biological fluids like CSF and urine for biomarker discovery, addressing sensitivity issues in complex samples.
Contribution
It details a complete method for preparing and analyzing dilute biological samples using 2D electrophoresis with fluorescence detection, enhancing proteomics biomarker research.
Findings
Effective sample concentration improves detection sensitivity.
Workflow enables reproducible analysis of dilute samples.
Suitable for biomarker discovery in less complex fluids.
Abstract
Two-dimensional electrophoresis is still a very valuable tool in proteomics, due to its reproducibility and its ability to analyze complete proteins. However, due to its sensitivity to dynamic range issues, its most suitable use in the frame of biomarker discovery is not on very complex fluids such as plasma, but rather on more proximal, simpler fluids such as CSF, urine, or secretome samples. Here, we describe the complete workflow for the analysis of such dilute samples by two-dimensional electrophoresis, starting from sample concentration, then the two-dimensional electrophoresis step per se, ending with the protein detection by fluorescence.
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