Syringe-push membrane absorption as a simple rapid method of urine preparation
for clinical proteomics
#MMPMID26074737
Chutipongtanate S
; Changtong C
; Weeraphan C
; Hongeng S
; Srisomsap C
; Svasti J
Clin Proteomics
2015[]; 12
(1
): 15
PMID26074737
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BACKGROUND: The 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. RESULT: Syringe-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. CONCLUSIONS: 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.
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