Applicability of 2D gel electrophoresis and liquid chromatography in proteomic analysis of urine using mass spectrometry MALDI-TOF

Banach, Tomasz; Adaszek, Łukasz; Wyłupek, D.; Winiarczyk, Mateusz; Winiarczyk, S.

doi:10.2478/pjvs-2013-0083

Cited by 4 publications

(2 citation statements)

References 46 publications

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“…Its name comes from a blend of proteins and genome. Proteomic analysis has become an important tool in biomedical and veterinary research [ 1 , 2 , 3 ]. The tear film covering the surface of the eye is a complex body fluid containing thousands of molecules with different structures and functions [ 3 – 7 ].…”

Section: Introductionmentioning

confidence: 99%

Dog Tear Film Proteome In-Depth Analysis

Winiarczyk

Banach

et al. 2015

PLoS ONE

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In this study, mass spectrometry was used to explore the canine tear proteome. Tear samples were obtained from six healthy dogs, and one-dimensional sodium dodecyl sulphate polyacrylamide gel electrophoresis (1D SDS-PAGE) was used as a first step to separate intact proteins into 17 bands. Each fraction was then trypsin digested and analysed by matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry (MALDI-TOF-MS/MS) to characterize the protein components in each fraction. In total, 125 tear proteins were identified, with MCA (Major Canine Allergen), Serum albumin, UPF0557 protein C10orf119 homolog, Collagen alpha-2(I) chain, Tyrosine -protein kinase Fer, Keratine type II cytoskeletal, Beta-crystallin B2, Interleukin-6 and Desmin occuring as the most confident ones with the highest scores. The results showed that the proteomic strategy used in this study was successful in the analysis of the dog tear proteome. To the best of our knowledge, this study is the first to report the comprehensive proteome profile of tears from healthy dogs by 1D SDS PAGE and MALDI-TOF. Data are available via ProteomeXchange with identifier PXD003124.

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Section: Introductionmentioning

confidence: 99%

Dog Tear Film Proteome In-Depth Analysis

Winiarczyk

Banach

et al. 2015

PLoS ONE

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“…Proteomics has become an important tool in biomedical and veterinary research [12,[14][15][16]. Despite well-developed veterinary ophthalmology research concerning dogs, reports on molecular studies of the tear film remains sparse.…”

Section: Introductionmentioning

confidence: 99%

Proteomic Analysis of Tear Film Obtained from Diabetic Dogs

Winiarczyk

et al. 2020

Animals

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Canine diabetes mellitus is a significant health burden, followed with numerous systemic complications, including diabetic cataracts and retinopathy, leading to blindness. Diabetes should be considered as a disease damaging all the body organs, including gastrointestinal tract, through a complex combination of vascular and metabolic pathologies, leading to impaired gut function. Tear film can be obtained in a non-invasive way, which makes it a feasible biomarker source. In this study we compared proteomic changes ongoing in tear film of diabetic dogs. The study group consisted of 15 diabetic dogs, and 13 dogs served as a control group. After obtaining tear film with Schirmer strips, we performed 2-dimensional electrophoresis, followed by Delta2D software analysis, which allowed to select statistically significant differentially expressed proteins. After their identification with MALDI-TOF (matrix assisted laser desorption and ionisation time of flight) spectrometry we found one up-regulated protein in tear film of diabetic dogs—SRC kinase signaling inhibitor 1 (SRCIN1). Eight proteins were down-regulated: phosphatidylinositol-4 kinase type 2 alpha (PI4KIIα), Pro-melanin concentrating hormone (Pro-MCH), Flotillin-1, Protein mono-ADP ribosyltransferase, GRIP and coiled coil domain containing protein 2, tetratricopeptide repeat protein 36, serpin, and Prelamin A/C. Identified proteins were analyzed by Panther Gene Ontology software, and their possible connections with diabetic etiopathology were discussed. We believe that this is the first study to target tear film proteome in canine diabetes. We believe that combined with traditional examination, the tear film proteomic analysis can be a new source of biomarkers both for clinical practice, and experimental research.

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Applications of MALDI-TOF mass spectrometry in clinical proteomics

Greco

Piras

Pieroni

et al. 2018

Expert Review of Proteomics

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The development of precision medicine requires advanced technologies to address the multifactorial disease stratification and to support personalized treatments. Among omics techniques, proteomics based on Mass Spectrometry (MS) is becoming increasingly relevant in clinical practice allowing a phenotypic characterization of the dynamic functional status of the organism. From this perspective, Matrix Assisted Laser Desorption Ionization Time of Flight (MALDI-TOF) MS is a suitable platform for providing a high-throughput support to clinics. Areas covered: This review aims to provide an updated overview of MALDI-TOF MS applications in clinical proteomics. The most relevant features of this analysis have been discussed, highlighting both pre-analytical and analytical factors that are crucial in proteomics studies. Particular emphasis is placed on biofluids proteomics for biomarkers discovery and on recent progresses in clinical microbiology, drug monitoring, and minimal residual disease (MRD). Expert commentary: Despite some analytical limitations, the latest technological advances together with the easiness of use, the low time and low cost consuming and the high throughput are making MALDI-TOF MS instruments very attractive for the clinical practice. These features offer a significant potential for the routine of the clinical laboratory and ultimately for personalized medicine.

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Applicability of 2D gel electrophoresis and liquid chromatography in proteomic analysis of urine using mass spectrometry MALDI-TOF

Cited by 4 publications

References 46 publications

Dog Tear Film Proteome In-Depth Analysis

Dog Tear Film Proteome In-Depth Analysis

Proteomic Analysis of Tear Film Obtained from Diabetic Dogs

Applications of MALDI-TOF mass spectrometry in clinical proteomics

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