Application of nano-LC-MALDI-TOF/TOF-MS for proteomic analysis of microvesicles

Kasprzyk, Joanna; Stępień, Ewa; Piekoszewski, Wojciech

doi:10.1016/j.clinbiochem.2016.11.013

Cited by 13 publications

(6 citation statements)

References 14 publications

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“…Filter aided sample preparation approach (FASP), a proteomic method for buffer exchange and protein digestion before MS analysis, and the related multiple enzyme digestion (MED) FASP have been successfully used in several EV studies, providing easy and efficient protein processing and minimal sample loss [ 123 , 124 ].…”

Section: Proteomic Methodsmentioning

confidence: 99%

Proteomics of Extracellular Vesicles: Update on Their Composition, Biological Roles and Potential Use as Diagnostic Tools in Atherosclerotic Cardiovascular Diseases

et al. 2020

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Extracellular vesicles (EVs) are lipid-bound vesicles released from cells under physiological and pathological conditions. Basing on biogenesis, dimension, content and route of secretion, they can be classified into exosomes, microvesicles (MVs) and apoptotic bodies. EVs have a key role as bioactive mediators in intercellular communication, but they are also involved in other physiological processes like immune response, blood coagulation, and tissue repair. The interest in studying EVs has increased over the years due to their involvement in several diseases, such as cardiovascular diseases (CVDs), and their potential role as biomarkers in diagnosis, therapy, and in drug delivery system development. Nowadays, the improvement of mass spectrometry (MS)-based techniques allows the characterization of the EV protein composition to deeply understand their role in several diseases. In this review, a critical overview is provided on the EV’s origin and physical properties, as well as their emerging functional role in both physiological and disease conditions, focusing attention on the role of exosomes in CVDs. The most important cardiac exosome proteomic studies will be discussed giving a qualitative and quantitative characterization of the exosomal proteins that could be used in future as new potential diagnostic markers or targets for specific therapies.

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Section: Proteomic Methodsmentioning

confidence: 99%

Proteomics of Extracellular Vesicles: Update on Their Composition, Biological Roles and Potential Use as Diagnostic Tools in Atherosclerotic Cardiovascular Diseases

et al. 2020

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“…Mean protein concentration was 1.14 ± 1.04 mg/mL; the total protein amount used for MS was 40 μ g. Proteomic analysis was performed by means of a nano-liquid chromatograph (EASY-nLC II™, Bruker Daltonics, Germany). The detailed methodology was previously published [20]. The precision tolerance was 100 ppm for peptide masses and 0.7 Da for fragment ion masses.…”

Section: Methodsmentioning

confidence: 99%

“…(a) Venn diagram shows that in EVs from controlled diabetic patient with microalbuminuria there is a large number of unique proteins ( n = 49), in a healthy control and in an uncontrolled diabetic patient with macroalbuminuria the number of unique proteins is very low ( n = 4) [20]. (b) Protein-to-protein interaction analysis of common 45 proteins selected from Venn diagram shows the central role of albumin among urinary EV-related proteins; the list of submitted proteins is available in a supplementary data file (Supplementary Table 2) [22].…”

Section: Figurementioning

confidence: 99%

Urinary Extracellular Vesicles: Potential Biomarkers of Renal Function in Diabetic Patients

Kamińska

Platt

Kasprzyk

et al. 2016

Journal of Diabetes Research

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The aim of this study was to check the relationship between the density of urinary EVs, their size distribution, and the progress of early renal damage in type 2 diabetic patients (DMt2). Patients were enrolled to this study, and glycated hemoglobin (HbA1c) below 7% was a threshold for properly controlled diabetic patients (CD) and poorly controlled diabetic patients (UD). Patients were further divided into two groups: diabetic patients without renal failure (NRF) and with renal failure (RF) according to the Glomerular Filtration Rate. Density and diameter of EVs were determined by Tunable Resistive Pulse Sensing. Additionally, EVs were visualized by means of Transmission and Environmental Scanning Electron Microscopy. Nano-liquid chromatography coupled offline with mass spectrometry (MALDI-TOF-MS/MS) was applied for proteomic analysis. RF had reduced density of EVs compared to NRF. The size distribution study showed that CD had larger EVs (mode) than UD (115 versus 109 nm; p < 0.05); nevertheless the mean EVs diameter was smaller in controls than in the CD group (123 versus 134 nm; p < 0.05). It was demonstrated that EVs are abundant in urine. Albumin, uromodulin, and number of unique proteins related to cell stress and secretion were detected in the EVs fraction. Density and size of urinary EVs reflect deteriorated renal function and can be considered as potential renal damage biomarkers.

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“…A shotgun proteomic approach by nanoLC-MS/MS and fractionation with hydrogel nanoparticles were applied to study proteomic profile of ADP-induced PMPs allowing the identification of about 600 proteins, among which the majority was distributed in the subsection cytoplasm and more than half had binding and catalytic activity [ 129 ]. Instead, Kasprzyk et al evaluated the protein composition of platelet microvesicles (PMVs) from thrombin-activated platelets of healthy donors using a nanoLC-MALDI-TOF/TOF MS analysis, and more than 400 proteins were identified, among which 123 were PMV-specific [ 151 ].…”

Section: Detailed Review Of the Literature Of Proteomics Of Platelmentioning

confidence: 99%

Platelets in Healthy and Disease States: From Biomarkers Discovery to Drug Targets Identification by Proteomics

Gianazza

Brioschi

Baetta

et al. 2020

IJMS

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Platelets are a heterogeneous small anucleate blood cell population with a central role both in physiological haemostasis and in pathological states, spanning from thrombosis to inflammation, and cancer. Recent advances in proteomic studies provided additional important information concerning the platelet biology and the response of platelets to several pathophysiological pathways. Platelets circulate systemically and can be easily isolated from human samples, making proteomic application very interesting for characterizing the complexity of platelet functions in health and disease as well as for identifying and quantifying potential platelet proteins as biomarkers and novel antiplatelet therapeutic targets. To date, the highly dynamic protein content of platelets has been studied in resting and activated platelets, and several subproteomes have been characterized including platelet-derived microparticles, platelet granules, platelet releasates, platelet membrane proteins, and specific platelet post-translational modifications. In this review, a critical overview is provided on principal platelet proteomic studies focused on platelet biology from signaling to granules content, platelet proteome changes in several diseases, and the impact of drugs on platelet functions. Moreover, recent advances in quantitative platelet proteomics are discussed, emphasizing the importance of targeted quantification methods for more precise, robust and accurate quantification of selected proteins, which might be used as biomarkers for disease diagnosis, prognosis and therapy, and their strong clinical impact in the near future.

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Application of nano-LC-MALDI-TOF/TOF-MS for proteomic analysis of microvesicles

Cited by 13 publications

References 14 publications

Proteomics of Extracellular Vesicles: Update on Their Composition, Biological Roles and Potential Use as Diagnostic Tools in Atherosclerotic Cardiovascular Diseases

Proteomics of Extracellular Vesicles: Update on Their Composition, Biological Roles and Potential Use as Diagnostic Tools in Atherosclerotic Cardiovascular Diseases

Urinary Extracellular Vesicles: Potential Biomarkers of Renal Function in Diabetic Patients

Platelets in Healthy and Disease States: From Biomarkers Discovery to Drug Targets Identification by Proteomics

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