Characterization of the bladder cancer urine proteome utilizing large dilution-reconcentration approach with liquid chromatography/mass spectrometry

摘要

Human urine contains a large of proteins and peptides (urinary proteome). Global analysis of human urinary proteome is important for understanding the urinary tract diseases. To enhance our understanding of urinary proteome of bladder cancer, the urine proteins were prepared by dilution-reconcentration sample cleanup preparation method and identified by nano-high performance liquid chromatography electrospray ionization tandem mass spectrometry (nano-HPLC-ESI-MS/MS) followed by peptide fragmentation patterns. The tangential flow filtration (TFF) system is useful for proteome sample purification and concentration may be specifically applied to urinary proteins to obtain detailed biochemical information on this unwieldy class of proteins. The experimental results demonstrated that a total of 466 unique peptides, corresponding to 410 unique proteins, were identified from bladder cancer urine samples. Among these 410 proteins identified, some proteins have not been reported as normal urinary proteins and may represent proteins specifically present in urine with bladder cancer. The proteomic analysis of urine samples is useful not only for the creating a proteomic map of urinary protein but also searching new protein markers of bladder cancer. This paper describes a strategy for utilizing a dilution-reconcentration approach (tangential flow filtration system, TFF system) as a urinary sample cleanup method for bladder cancer urinary proteome analysis by nano-high performance liquid chromatography electrospray ionization tandem mass spectrometry (nano-HPLC-ESI-MS/MS). TFF system is useful for proteome sample purification and concentration may be specifically applied to urinary proteins to obtain detailed biochemical information on this unwieldy class of proteins.