Capillary zone electrophoresis of bacterial extracellular vesicles: A proof of concept

Piotrowska, Martyna; Ciura, Krzesimir; Zalewska, Michalina; Dawid, Marta; Correia, Bruna; Sawicka, Paulina; Lewczuk, Bogdan; Kasprzyk, Joanna; Solá, Laura; Piekoszewski, Wojciech; Wielgomas, Bartosz; Waleron, Krzysztof; Dziomba, Szymon

doi:10.1016/j.chroma.2020.461047

Cited by 33 publications

(63 citation statements)

References 59 publications

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“…Furthermore, unsatisfactory EVs recovery (less than 60%) was obtained with the centrifugal filtration approach, compared to that achieved with the Spin Column-based alternative (75 % recovery, with deviation less than 5 % for a repeatability test on 4 EV samples), presumably due to EVs sticking to the NanoSep filter membranes [36]. A similar observation on low EVs recovery and reproducibility after filtration was also made in a recent work on CE-UV separation of EVs [18].…”

Section: Matrix Removal Strategy -17 -supporting

confidence: 66%

“…Interestingly, this observation was also reported in the recently released work on CE-UV of EVs, in which large ions (i.e. bis-tris propane ions) were employed to maintain the EV signal stability [18]. The ISF BGEs, which are tolerant to the presence of PBS in the sample matrix, were found to minimize protein adsorption to capillary wall and induce excellent stacking of slowly migrating proteins.…”

Section: Ce-lif Of Fluorescently Labelled Evssupporting

confidence: 57%

“…This common problem observed with nanoparticles was also observed during the CE of liposomes [34]. Interestingly, another recent work on CE-UV of EVs also revealed the presence of spikes during electrophoresis [18]. Since EVs suspended in PBS were injected to the CE-LIF, the presence of -16 -PBS in the sample plugs (accounting for 10 % of the total capillary volume) are likely to produce local Joule heating under a high electrical field due to its high conductivity.…”

Section: Ce-lif Of Fluorescently Labelled Evsmentioning

confidence: 62%

“…-5 -After the submission of our manuscript on CE-LIF of EVs, another work on CE-UV of EVs was published [18]. Together with this pioneering work, we provide herein the first proof of concept on electrokinetic separation and characterization of EVs.…”

Section: Introductionmentioning

confidence: 82%

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Electrokinetic characterization of extracellular vesicles with capillary electrophoresis: A new tool for their identification and quantification

Morani

Duc

Krupová³

et al. 2020

Analytica Chimica Acta

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This work reports on the development of the first capillary electrophoresis methodology for the elucidation of extracellular vesicles' (EVs) electrokinetic distributions. The approach is based on capillary electrophoresis coupled with laser-induced fluorescent (LIF) detection for the identification and quantification of EVs after their isolation. Sensitive detection of these nanometric entities was possible thanks to an 'inorganic-species-free' background electrolyte.This electrolyte was made up of weakly charged molecules at very high concentrations to stabilize EVs, and an intra-membrane labelling approach was used to prevent EV morphology modification. The limit of detection for EVs achieved using the developed CE-LIF method reached 8 × 10 9 EVs / mL, whereas the calibration curve was acquired from 1.22 × 10 10 to 1.20 × 10 11 EVs / mL. The CE-LIF approach was applied to provide the electrokinetic distributions of various EVs of animal and human origins, and visualize different EV subpopulations from our recently developed high-yield EV isolation method.

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Section: Matrix Removal Strategy -17 -supporting

confidence: 66%

Section: Ce-lif Of Fluorescently Labelled Evssupporting

confidence: 57%

Section: Ce-lif Of Fluorescently Labelled Evsmentioning

confidence: 62%

Section: Introductionmentioning

confidence: 82%

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Electrokinetic characterization of extracellular vesicles with capillary electrophoresis: A new tool for their identification and quantification

Morani

Duc

Krupová³

et al. 2020

Analytica Chimica Acta

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“…Capillary electrophoresis can separate molecules or entities based on the differences in the charge-mass ratio of single molecules, complexes of molecules, circulating cells, biological particles, and even man-made nanoscale quantum dots [ 215 ]. Recently, it has been proposed as a proof of concept that capillary electrophoresis can be used for the separation of bacterial extracellular vesicles [ 216 ]. Much effort has been performed during the last 5 years to develop methods for the isolation and characterization of exosomes, which are seen as attractive candidates for clinical application as innovative diagnostic and therapeutic tools [ 217 , 218 , 219 , 220 , 221 , 222 , 223 , 224 ].…”

Section: The Potential Use Of Iace For Extracellular Vesicle Studimentioning

confidence: 99%

A Two-Dimensional Affinity Capture and Separation Mini-Platform for the Isolation, Enrichment, and Quantification of Biomarkers and Its Potential Use for Liquid Biopsy

Guzman¹,

Guzman

2020

Biomedicines

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Biomarker detection for disease diagnosis, prognosis, and therapeutic response is becoming increasingly reliable and accessible. Particularly, the identification of circulating cell-free chemical and biochemical substances, cellular and subcellular entities, and extracellular vesicles has demonstrated promising applications in understanding the physiologic and pathologic conditions of an individual. Traditionally, tissue biopsy has been the gold standard for the diagnosis of many diseases, especially cancer. More recently, liquid biopsy for biomarker detection has emerged as a non-invasive or minimally invasive and less costly method for diagnosis of both cancerous and non-cancerous diseases, while also offering information on the progression or improvement of disease. Unfortunately, the standardization of analytical methods to isolate and quantify circulating cells and extracellular vesicles, as well as their extracted biochemical constituents, is still cumbersome, time-consuming, and expensive. To address these limitations, we have developed a prototype of a portable, miniaturized instrument that uses immunoaffinity capillary electrophoresis (IACE) to isolate, concentrate, and analyze cell-free biomarkers and/or tissue or cell extracts present in biological fluids. Isolation and concentration of analytes is accomplished through binding to one or more biorecognition affinity ligands immobilized to a solid support, while separation and analysis are achieved by high-resolution capillary electrophoresis (CE) coupled to one or more detectors. When compared to other existing methods, the process of this affinity capture, enrichment, release, and separation of one or a panel of biomarkers can be carried out on-line with the advantages of being rapid, automated, and cost-effective. Additionally, it has the potential to demonstrate high analytical sensitivity, specificity, and selectivity. As the potential of liquid biopsy grows, so too does the demand for technical advances. In this review, we therefore discuss applications and limitations of liquid biopsy and hope to introduce the idea that our affinity capture-separation device could be used as a form of point-of-care (POC) diagnostic technology to isolate, concentrate, and analyze circulating cells, extracellular vesicles, and viruses.

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Molecular Separation by Using Active and Passive Microfluidic chip Designs: A Comprehensive Review

Ebrahimi,

Icoz,

Didarian

et al. 2023

Adv Materials Inter

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Separation and identification of molecules and biomolecules such as nucleic acids, proteins, and polysaccharides from complex fluids are known to be important due to unmet needs in various applications. Generally, many different separation techniques, including chromatography, electrophoresis, and magnetophoresis, have been developed to identify the target molecules precisely. However, these techniques are expensive and time consuming. “Lab‐on‐a‐chip” systems with low cost per device, quick analysis capabilities, and minimal sample consumption seem to be ideal candidates for separating particles, cells, blood samples, and molecules. From this perspective, different microfluidic‐based techniques have been extensively developed in the past two decades to separate samples with different origins. In this review, “lab‐on‐a‐chip” methods by passive, active, and hybrid approaches for the separation of biomolecules developed in the past decade are comprehensively discussed. Due to the wide variety in the field, it will be impossible to cover every facet of the subject. Therefore, this review paper covers passive and active methods generally used for biomolecule separation. Then, an investigation of the combined sophisticated methods is highlighted. The spotlight also will be shined on the elegance of separation successes in recent years, and the remainder of the article explores how these permit the development of novel techniques.

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Capillary zone electrophoresis of bacterial extracellular vesicles: A proof of concept

Cited by 33 publications

References 59 publications

Electrokinetic characterization of extracellular vesicles with capillary electrophoresis: A new tool for their identification and quantification

Electrokinetic characterization of extracellular vesicles with capillary electrophoresis: A new tool for their identification and quantification

A Two-Dimensional Affinity Capture and Separation Mini-Platform for the Isolation, Enrichment, and Quantification of Biomarkers and Its Potential Use for Liquid Biopsy

Molecular Separation by Using Active and Passive Microfluidic chip Designs: A Comprehensive Review

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