Optimization of analytical and pre-analytical conditions for MALDI-TOF-MS human urine protein profiles

Calvano, Cosima Damiana; Aresta, Antonella; Iacovone, M.; Benedetto, Giuseppe E. De; Zambonin, Carlo; Battaglia, Michele; Ditonno, P.; Rutigliano, Monica; Bettocchi, Carlo

doi:10.1016/j.jpba.2009.10.014

Cited by 36 publications

(31 citation statements)

References 27 publications

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“…28 Microparticles in the supernatant can be isolated for further analyses by ultracentrifugation at a later stage, after storage at -80°C. 21,36 Urine preanalytical conditions have been studied in specific proteomic methods including MALDI-TOF-MS, 37 2D-electrophoresis, 4 SELDITOF MS, 31 and LC-MS, 38 showing that collection and processing variables can have a major impact on specific proteins and peptides. In this context, laboratories and biobanks operating under accreditation are advised to track and record preanalytical data systematically using the Standard PREanalytical Code (SPREC), 39,40 while freeze-thaw cycles and other treatments (e.g., use of protease inhibitors, stabilizers) should be reported in the Laboratory Information Management System.…”

Section: Urine Processing Methods 355 Discussionmentioning

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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Section: Urine Processing Methods 355 Discussionmentioning

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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“…The changes of protein (Panicker et al, 2009;Calvano et al, 2010) were detected in the cervix mucus protein of patients with cervical cancer and urine of prostate cancer patients. 2) Small sample volume and prompt detection.…”

Section: Discussionmentioning

confidence: 99%

Detection and Prognostic Analysis of Serum Protein Expression in Esophageal Squamous Cell Cancer

Jiang¹,

Wang²,

Yu³

et al. 2012

Asian Pacific Journal of Cancer Prevention

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Objective: To assess differences in serum proteins in esophageal squamous cell carcinoma patients. Methods: 144 esophageal squamous cell carcinoma patients and 50 healthy volunteers were included in this study, with surface-enhanced laser desorption-ionization time-of-flight mass spectrometry and weak cation exchange magnetic beads. Follow-up allowed the relations between serum proteins and prognosis to be analyzed. Results: A total of 93 protein peaks were detected (molecular weight range: 1500-30000), 10 demonstrating statistically significant differences. There were no differences in protein peaks between 92 patients with a survival more than 2 years and 52 patients with survival less than 2 years. There were two significantly different protein peaks between 45 stage Ⅱ patients with a survival more than 2 years and 14 stage Ⅱ patients with survival less than 2 years. There was one significantly different protein peak between 22 stage Ⅲ patients with a survival more than 2 years and 29 stage Ⅲ patients with survival less than 2 years. Conclusion: Differences of serum proteins in esophageal squamous cell carcinoma are related to prognosis of patients. The protein fingerprint can be helpful for clinical diagnosis and treatment.

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“…9,10 Among the different types of experimental variability, preanalytical variations, including pre-and post-sample acquisition, are often not appreciated and thus not controlled. 11,12 Preanalytical processes are defined as those procedures taking place between specimen collection and experimental assay/analysis. Even small differences in SOPs between groups or within groups could yield uninterpretable results due to variations at multiple steps in the collection and handling process.…”

Section: Standardization and Best Practices Guidelinesmentioning

confidence: 99%

Enhancing translational research in paediatric rheumatology through standardization

et al. 2016

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Optimization of analytical and pre-analytical conditions for MALDI-TOF-MS human urine protein profiles

Cited by 36 publications

References 27 publications

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

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

Detection and Prognostic Analysis of Serum Protein Expression in Esophageal Squamous Cell Cancer

Enhancing translational research in paediatric rheumatology through standardization

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