Dataset Information

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

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.

SUBMITTER: Chutipongtanate S

PROVIDER: S-EPMC4464716 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature

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

Chutipongtanate Somchai S Changtong Channarong C Weeraphan Churat C Hongeng Suradej S Srisomsap Chantragan C Svasti Jisnuson J

Clinical proteomics 20150606 1

<h4>Background</h4>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.<h4>Result</h4>Syringe-push membrane absorption (SPMA) was successfully developed by a combination of 5-ml medical syringe and ...[more]

PMID: 26074737

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Project description:BACKGROUND:Carprofen and platelet-rich plasma (PRP) are widely used in small animal clinical practice. Separation layers have been used during blood centrifugation to increase platelet yield. The objectives of this study were to (1) identify the optimal centrifugation force for the one-step PRP preparation, (2) determine whether there is an advantage to using carprofen in one-step PRP preparation, and (3) compare platelet morphology from one-step PRP preparation with and without carprofen. We hypothesized that injectable carprofen (emulsion formula) could be used successfully as the separation layer in PRP preparation. RESULTS:Samples from 14 healthy dogs were used to determine the optimal centrifugation force using one-step PRP preparation in a disposable syringe without carprofen, with forces set at 300, 500, 700, 900, 1100, 1300, and 1500 xg for 5?min. Optimum centrifugation force, plasma volume, and platelet concentrations of one-step PRP preparation were found and recovered at 900 xg, 1.9?±?0.28?ml, and 260.50?±?58.39 X 103 cell/?l, respectively. Samples from 12 healthy dogs were used to determine the optimal force (with forces set at 300, 500, 700, and 900 xg) for 5?min using one-step PRP preparation with carprofen. Optimum centrifugation force, plasma volume, and platelet concentrations for one-step PRP preparation with carprofen were found and recovered at 500 xg, 0.62?±?0.16?ml and 948.50?±?261.40 X 103 cell/?l, respectively. One-step PRP preparation with carprofen increased the platelet yield from baseline by 1.76 and 4.95 fold, respectively. Samples from 3 healthy dogs were used to observe platelet morphologies after centrifugation by scanning electron microscopy. Images of platelets on glass slides from both preparation methods revealed pseudopods emerging from the margins of the discoid platelets. CONCLUSIONS:One-step PRP centrifugation both with and without carprofen increased the platelet yield, but using carprofen (emulsion formula) as a separation layer resulted in a higher platelet yield. The clinical usefulness of PRP products from these methods should be further investigated.

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Dataset Information

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

Background

Result

Conclusions

Publications

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

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