Syringe-push membrane absorption as a simple rapid method of urine preparation for clinical proteomics

Clinical Proteomics

Somchai Chutipongtanate¹  Churat Weeraphan³  Chantragan Srisomsap³  Suradej Hongeng⁴  Channarong Changtong⁴  Jisnuson Svasti² 

[1] Central Laboratory, Department of Pediatrics, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, ThailandCentral Laboratory, Department of Pediatrics, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, ThailandCentral Laboratory, Department of Pediatrics, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, Thailand;Laboratory of Biochemistry, Chulabhorn Research Institute, Bangkok, ThailandApplied Biological Sciences Program, Chulabhorn Graduate Institute, Bangkok, ThailandLaboratory of Biochemistry, Chulabhorn Research Institute, Bangkok, ThailandLaboratory of Biochemistry, Chulabhorn Research Institute, Bangkok, ThailandApplied Biological Sciences Program, Chulabhorn Graduate Institute, Bangkok, ThailandApplied Biological Sciences Program, Chulabhorn Graduate Institute, Bangkok, ThailandLaboratory of Biochemistry, Chulabhorn Research Institute, Bangkok, ThailandApplied Biological Sciences Program, Chulabhorn Graduate Institute, Bangkok, Thailand;Laboratory of Biochemistry, Chulabhorn Research Institute, Bangkok, ThailandLaboratory of Biochemistry, Chulabhorn Research Institute, Bangkok, ThailandLaboratory of Biochemistry, Chulabhorn Research Institute, Bangkok, Thailand;Hematology and Oncology Unit, Department of Pediatrics, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, ThailandHematology and Oncology Unit, Department of Pediatrics, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, ThailandHematology and Oncology Unit, Department of Pediatrics, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, Thailand

关键词: Comparison;    Membrane absorption;    Proteomics;    Urine preparation;   

DOI  :  10.1186/s12014-015-9087-4

来源: Humana Press Inc

【 摘 要 】

Abstract

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.

【 授权许可】

Unknown