Urine Collection and Processing for Protein Biomarker Discovery and Quantification

Thomas, Charles E.; Sexton, Wade J.; Benson, Kaaron; Sutphen, Rebecca; Koomen, John M.

doi:10.1158/1055-9965.epi-10-0069

Cited by 117 publications

(94 citation statements)

References 61 publications

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“…It can be obtained by noninvasive collection methods and thus is more easily collected than the tissue or blood, which enables the more convenient monitoring of a wide range of physiologic processes and diseases. Moreover, urine aliquots provide enough protein for multiple analyses (51,52). …”

Section: Introductionmentioning

confidence: 99%

Histidine Decarboxylase Is Identified as a Potential Biomarker of Intestinal Mucosal Injury in Patients with Acute Intestinal Obstruction

Yang

Zhāng

et al. 2011

Mol Med

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Various biomarkers currently used for the diagnosis of intestinal mucosal injury (IMI) in patients with acute intestinal obstruction have low sensitivity and specificity. In the present study, IMI, as indicated by the impaired expression of tight junction proteins, including zonula occludens-1, occludin and claudin-1, and inflammation were determined in colonic tissues of patients with 45 strangulated intestinal obstruction (STR-IO) and the adjacent "normal" colonic tissues of 35 patients with colon cancers by quantitative real-time polymerase chain reaction (QRT-PCR), Western blotting, immunohistochemistry and histological examination, respectively. Then, two-dimensional fluorescent difference gel electrophoresis coupled with linear trap quadrupole mass spectrometry was used to screen for potential biomarkers of IMI in the serum samples of 10 STR-IO, 10 simple intestinal obstruction (SIM-IO) and 10 normal healthy controls. A total of 35 protein spots were differentially expressed among the serum samples, and six of the proteins were identified as potential biomarkers. Among the six proteins, histidine decarboxylase (HDC) and ceruloplasmin (CP) were elevated significantly in patients with STR-IO, compared with patients with SIM-IO and healthy controls. Thus, HDC and CP were further validated by QRT-PCR, Western blotting, immunohistochemistry and enzyme-linked immunosorbent assay, respectively, in colonic tissues, serum and urine samples. Finally, the receiver operating characteristic curves were used to show the area under the curves of HDC, CP and several established biomarkers, followed by the determination of the appropriate cutoff values and their sensitivities and specificities. It was shown that for serum and urine, HDC levels achieved sensitivities and specificities compatible to or even greater than those of established biomarkers for the diagnosis of IMI in patients with acute intestinal obstruction, although further validation in a larger cohort is required.

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Section: Introductionmentioning

confidence: 99%

Histidine Decarboxylase Is Identified as a Potential Biomarker of Intestinal Mucosal Injury in Patients with Acute Intestinal Obstruction

Yang

Zhāng

et al. 2011

Mol Med

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“…Urine is proving to be an increasingly useful source of metabolites and protein biomarkers for detecting disease states in both adults and neonates, [4][5][6][7] as well as the effects of drugs and dietary intake on physiologic processes. [8][9][10] Best Practices 11 and Standard Operating Procedures [12][13][14][15][16] for routine processing and storing of biobank samples have been published.…”

mentioning

confidence: 99%

Method Validation for Preparing Urine Samples for Downstream Proteomic and Metabolomic Applications

Ammerlaan

Trezzi

Mathay

et al. 2014

Biopreservation and Biobanking

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Background: Formal validation of methods for biospecimen processing in the context of accreditation in laboratories and biobanks is lacking. A protocol for processing of a biospecimen (urine) was validated for fitness-for-purpose in terms of key downstream endpoints. Methods: Urine processing was optimized for centrifugation conditions on the basis of microparticle counts at room temperature (RT) and at 4°C. The optimal protocol was validated for performance (microparticle counts), and for reproducibility and robustness for centrifugation temperature (4°C vs. RT) and brake speed (soft, medium, hard). Acceptance criteria were based on microparticle counts, cystatin C and creatinine concentrations, and the metabolomic profile. Results: The optimal protocol was a 20-min, 12,000 g centrifugation at 4°C, and was validated for urine collection in terms of microparticle counts. All reproducibility acceptance criteria were met. The protocol was robust for centrifugation at 4°C versus RT for all parameters. The protocol was considered robust overall in terms of brake speeds, although a hard brake gave significantly fewer microparticles than a soft brake. Conclusions: We validated a urine processing method suitable for downstream proteomic and metabolomic applications. Temperature and brake speed can influence analytic results, with 4°C and high brake speed considered optimal. Laboratories and biobanks should ensure these conditions are systematically recorded in the scope of accreditation.

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“…We specifically chose to use strict entry criteria for this pilot study to show proof of concept and obtain preliminary data to justify inclusion into a clinical trial. Additionally, urinary biomarkers can indicate substantial variability from day to day, hydration status, and a multitude of other factors [22]. We limited the evaluation of urinary biomarkers for post prostatectomy solely to MCP-1.…”

Section: Discussionmentioning

confidence: 99%

Monocyte Chemotactic Protein-1 (MCP-1) as a Predictor of Prolonged Urinary Incontinence After Radical Prostatectomy

Liss¹,

Ahlering

Morales

et al. 2016

TOUNJ

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Objectives:To investigate monocyte chemotactic protein-1 (MCP-1) as a novel urinary biomarker to predict prolonged post prostatectomy incontinence. Methods:Men submitted urine samples prior to robotic radical prostatectomy. MCP-1 values were derived using an ELISA test. Pad usage at 7, 30, and 60 days were documented by patient post cards mailed when zero pads was reached. The primary outcome was defined as no incontinence pad usage at 30 days at prostatectomy. Results:After exclusions, 76 patients were included in analyses. Continence was reached by 29% (22/76), 56% (42/76), and (75/76) 98% at 7, 30, and 60 days, respectively. The average MCP-1 (p=0.258) was not different between the continent and incontinent groups. Highest quartile of MCP-1 (MCP > 166 pg/mL) and normalized MCP-1 (MCP-1/TV >0.53) noted a significant delay in continence at 30 days (p=0.050 and p=0.003). Only 26% (5/19) in the highest MCP1/TV quartile were continent, whereas 65% (37/57) of men in the 3 lower quartiles reached zero pad continence (p=0.003). In a logistic regression model the highest quartile of MCP1/TV had a significant chance of being incontinent at 30 days (OR 0.22; 95% CI 0.058-0.80; p=0.022). Conclusion:MCP-1/TV is a urinary biomarker that may predict prolonged urinary incontinence after radical prostatectomy.

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Urine Collection and Processing for Protein Biomarker Discovery and Quantification

Cited by 117 publications

References 61 publications

Histidine Decarboxylase Is Identified as a Potential Biomarker of Intestinal Mucosal Injury in Patients with Acute Intestinal Obstruction

Histidine Decarboxylase Is Identified as a Potential Biomarker of Intestinal Mucosal Injury in Patients with Acute Intestinal Obstruction

Method Validation for Preparing Urine Samples for Downstream Proteomic and Metabolomic Applications

Monocyte Chemotactic Protein-1 (MCP-1) as a Predictor of Prolonged Urinary Incontinence After Radical Prostatectomy

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