MALDI-TOF/MS Analysis of Extracellular Vesicles Released by Cancer Cells

Zambonin, Carlo

doi:10.3390/app12126149

Cited by 7 publications

(4 citation statements)

References 55 publications

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“…EVs should be classi ed based on their characteristics, as well as their prognostic, diagnostic, and clinical requirements, as well as their cost-effectiveness (Van der Meel et al, 2014). High selectivity and high e ciency are the main areas to focus on when it comes to isolation and puri cation (Zambonin, 2022). It is essential that the vesicles and their contents are characterized in depth and that the isolation processes are standardized before they can be used (Tian et al, 2022).…”

Section: Background Informationmentioning

confidence: 99%

Isolation of extracellular vesicles from pine (Pinus sp.) leaves and roots

Kunene

Steenkamp

Motaung

2023

Preprint

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Plants release extracellular vesicles (EVs) for various reasons, such as unconventional protein secretion, RNA transport, and defence against pathogens. However, very little is known about plant EV biosynthesis and diverse and bioactive luminal contents, including how they alter recipient cell biology and function. To study EVs, researchers often isolate them from apoplastic wash or juice fluids using various methods. A method is needed to isolate EVs from non-model plants with tougher leaves such as species in the Pinus genus. Because of their spindly and sharp shape, pines have strong acicular-shaped leaves that are commonly called needles. Unlike all other taproots, pine seedlings grow into a primary root first, which gives rise to lateral roots later. There are also various dimensions and shapes of pine needles, but they are relatively strong compared to the leaves of other plants such as Arabidopsis thaliana, where most EV studies have been concentrated. Extracting EVs from a juice fluid is highly effective for plants with tougher leaves because it ensures that the cells have been broken down and the vesicles are present. Given these reasons, we describe our method for the isolation and purification of EVs from needles and roots of different pine species. A detailed protocol is provided that can guide the collection of juice fluid from pine needles and roots in a clean and efficient manner. A number of other plant species with similar morphologies can greatly benefit from the use of this protocol. Basic protocol 1: Isolation of extracellular vesicles from juice fluid of pine leaves and roots Basic protocol 2: Isolation of pine extracellular vesicles from juice fluid using size exclusion chromatography

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Section: Background Informationmentioning

confidence: 99%

Isolation of extracellular vesicles from pine (Pinus sp.) leaves and roots

Kunene

Steenkamp

Motaung

2023

Preprint

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“…Quantifications of CDK 4/6 inhibitors (with major metabolites of the drug) and PARP inhibitors from dried blood spots (having implications of avoiding phlebotomy) have recently been reported. Two reviews by Zambonin (2022) highlight the use of MALDI-TOF/MS for biomarker identification . One on cancer released extracellular vesicular proteins and proteins from urine and saliva samples was the first review.…”

Section: Illustrations Of Different Mass Spectrometry Instrumentation...mentioning

confidence: 99%

“…Two reviews by Zambonin (2022) highlight the use of MALDI-TOF/MS for biomarker identification. 88 One on cancer released extracellular vesicular proteins and proteins from urine and saliva samples was the first review. The second review provides an overview of recent applications and highlights the potential of MALDI-ToF/MS in identifying cancer metabolic biomarkers from body fluids.…”

Section: Lc-ms/msmentioning

confidence: 99%

Recent Contributions of Mass Spectrometry-Based “Omics” in the Studies of Breast Cancer

Banerjee,

Hatimuria,

Sarkar

et al. 2023

Chem. Res. Toxicol.

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Breast cancer (BC) is one of the most heterogeneous groups of cancer. As every biotype of BC is unique and presents a particular "omic" signature, they are increasingly characterized nowadays with novel mass spectrometry (MS) strategies. BC therapeutic approaches are primarily based on the two features of human epidermal growth factor receptor 2 (HER2) and estrogen receptor (ER) positivity. Various strategic MS implementations are reported in studies of BC also involving data independent acquisitions (DIAs) of MS which report novel differential proteomic, lipidomic, proteogenomic, phosphoproteomic, and metabolomic characterizations associated with the disease and its therapeutics. Recently many "omic" studies have aimed to identify distinct subsidiary biotypes for diagnosis, prognosis, and targets of treatment. Along with these, drug-induced-resistance phenotypes are characterized by "omic" changes. These identifying aspects of the disease may influence treatment outcomes in the near future. Drug quantifications and characterizations are also done regularly and have implications in therapeutic monitoring and in drug efficacy assessments. We report these studies, mentioning their implications toward the understanding of BC. We briefly provide the MS instrumentation principles that are adopted in such studies as an overview with a brief outlook on DIA-MS strategies. In all of these, we have chosen a model cancer for its revelations through MS-based "omics".

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“…MALDI is well suited for clinical practice, as it is user-friendly even for unskilled personnel, allowing for the rapid generation of spectra and profiles from the matrix under study. Particularly, the potential, limitations, and potential implementation strategies of MALDI analysis of exosomes have recently been thoroughly discussed [19], while applications of MALDI to the analysis of the EVs secreted by cancer cells have been reviewed [20]. Given the considerable and growing number of existing works focusing on MALDI characterization of the EVs released by human cells in various pathological "non-tumor" conditions, as well as by animal cells and microorganisms, this work aims to comprehensively review the related literature.…”

Section: Introductionmentioning

confidence: 99%

Analysis and Characterization of the Extracellular Vesicles Released in Non-Cancer Diseases Using Matrix-Assisted Laser Desorption Ionization/Mass Spectrometry

Aresta,

De Vietro,

Zambonin

2024

IJMS

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The extracellular vesicles (EVs) released by cells play a crucial role in intercellular communications and interactions. The direct shedding of EVs from the plasma membrane represents a fundamental pathway for the transfer of properties and information between cells. These vesicles are classified based on their origin, biogenesis, size, content, surface markers, and functional features, encompassing a variety of bioactive molecules that reflect the physiological state and cell type of origin. Such molecules include lipids, nucleic acids, and proteins. Research efforts aimed at comprehending EVs, including the development of strategies for their isolation, purification, and characterization, have led to the discovery of new biomarkers. These biomarkers are proving invaluable for diagnosing diseases, monitoring disease progression, understanding treatment responses, especially in oncology, and addressing metabolic, neurological, infectious disorders, as well as advancing vaccine development. Matrix-Assisted Laser Desorption Ionization (MALDI)/Mass Spectrometry (MS) stands out as a leading tool for the analysis and characterization of EVs and their cargo. This technique offers inherent advantages such as a high throughput, minimal sample consumption, rapid and cost-effective analysis, and user-friendly operation. This review is mainly focused on the primary applications of MALDI–time-of-flight (TOF)/MS in the analysis and characterization of extracellular vesicles associated with non-cancerous diseases and pathogens that infect humans, animals, and plants.

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MALDI-TOF/MS Analysis of Extracellular Vesicles Released by Cancer Cells

Cited by 7 publications

References 55 publications

Isolation of extracellular vesicles from pine (Pinus sp.) leaves and roots

Isolation of extracellular vesicles from pine (Pinus sp.) leaves and roots

Recent Contributions of Mass Spectrometry-Based “Omics” in the Studies of Breast Cancer

Analysis and Characterization of the Extracellular Vesicles Released in Non-Cancer Diseases Using Matrix-Assisted Laser Desorption Ionization/Mass Spectrometry

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