Abstract
The proteomic analysis of serum (plasma) has been a major approach to determining biomarkers essential for early disease diagnoses and drug discoveries. The determination of these biomarkers, however, is analytically challenging since the dynamic concentration range of serum proteins/peptides is extremely wide (more than 10 orders of magnitude). Thus, the reduction in sample complexity prior to proteomic analyses is essential, particularly in analyzing lowly abundant protein biomarkers. Here, we demonstrate a novel approach to the proteomic analyses of human serum that uses an originally developed serum protein separation device and a subsequently linked high-performance mass spectrometer system. The hollow-fiber membrane based serum pre-treatment device we developed can efficiently deplete high-molecular-weight proteins and concentrate low-molecular-weight proteins/peptides automatically within an hour. The proteomic analysis of healthy human serum pre-treated using the device, followed by the mass spectrometer, successfully identified about 2,000 proteins. According to the concept of the pre-treatment device, 64% of the identified proteins were smaller proteins than the human serum albumin. We believe this unique serum pre-treatment device and the proteomic analysis protocol allow high-throughput and efficient discovery of serum disease biomarkers.
