Abstract
BackgroundThe analysis of urinary proteome might reveal biomarkers of clinical value. However, current methods of urine preparation for down-stream proteomic analysis are complicated, time-consuming, and/or expensive. This study aims to develop a robust, simple, inexpensive and readily accessible urine preparation method to facilitate clinical proteomic workflow.ResultSyringe-push membrane absorption (SPMA) was successfully developed by a combination of 5-ml medical syringe and protein-absorbable membrane. Comparing three membranes i.e., nitrocellulose, polyvinylidene difluoride (PVDF) and Whatman no.1, nitrocellulose combined with SPMA (nitrocellulose-SPMA) provided the greatest quality of proteome profile as demonstrated by 2-DE. The quality of the proteome profile and the performance of nitrocellulose-SPMA were systematically compared with three current methods of urine preparation (i.e., ultrafiltration, dialysis/lyophilization and precipitation). While different methods of urine preparation provided comparable proteome quality, nitrocellulose-SPMA had better working performance due to acceptable recovery yield, less workload, short working time, high accessibility and low unit cost. In addition, protein absorbed on nitrocellulose harvested from the SPMA procedure could be stored as a dried membrane at room temperature for at least 1-month without protein degradation or modification.ConclusionsSPMA is a simple rapid method of preparing urine for downstream proteomic analysis. Because of it is highly accessible and has long storage duration, this technique holds potential benefit for large-scale multi-center research and future development of clinical investigation based upon urinary proteomic analysis.Electronic supplementary materialThe online version of this article (doi:10.1186/s12014-015-9087-4) contains supplementary material, which is available to authorized users.
Highlights
The analysis of urinary proteome might reveal biomarkers of clinical value
Syringe-push membrane absorption (SPMA) is a simple rapid method of preparing urine for downstream proteomic analysis. Because of it is highly accessible and has long storage duration, this technique holds potential benefit for large-scale multi-center research and future development of clinical investigation based upon urinary proteomic analysis
Establishment of SPMA The rationale of SPMA development was to devise a simple method of urine preparation that can be performed anywhere and is suitable for large-scale multi-center analysis, while preserving the best quality of urinary proteins for downstream proteomic analysis
Summary
The analysis of urinary proteome might reveal biomarkers of clinical value. current methods of urine preparation for down-stream proteomic analysis are complicated, time-consuming, and/or expensive. Urine contains a relatively low amount of proteins, and is contaminated with a large amount of waste products Those techniques provide a high quality urinary proteome profile. A new method of urine preparation that is uncomplicated, robust, inexpensive and readily accessible would facilitate large-scale multi-center analysis
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