Biased cyclical electrical field-flow fractionation for separation of submicron particles

Ornthai, Mathuros; Siripinyanond, Atitaya; Gale, Bruce K.

doi:10.1007/s00216-015-9173-5

Cited by 11 publications

(12 citation statements)

References 14 publications

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“…The ElFFF device used in the characterization tests was the same device as described in previous publications from our group. ,− The ElFFF channel dimensions are 64 cm length, 178 μm height, and 2 cm breadth. A high-pressure isocratic pump (Agilent model 1260, Agilent, Inc., Santa Clara, CA) pumped the carrier liquid through the ElFFF system.…”

Section: Methodsmentioning

confidence: 99%

“…ElFFF is a technique that separates particles based upon their electrophoretic mobility. This technique has been used for the characterization and separation of nanoparticles. ,− In this work, the ability of ElFFF to separate human plasma and U87 sEVs and mEVs was investigated, and their behavior was studied under different electrical conditions. Further, cyclical ElFFF (CyElFFF) theory was applied to calculate the electrophoretic mobility of U87 sEVs using fractionation data.…”

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confidence: 99%

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Characterization of Human Glioblastoma versus Normal Plasma-Derived Extracellular Vesicles Preisolated by Differential Centrifugation Using Cyclical Electrical Field-Flow Fractionation

et al. 2020

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Although many properties for small extracellular vesicles (sEVs, formerly termed “exosomes”) isolated at ∼100 000g are known, a wide range of values are reported for their electrophoretic mobility (EM) measurements. This paper reports for the first time the effect of dilution on the EM of U87 glioblastoma cell-derived and plasma-derived sEVs and medium size EVs (mEVs, commonly termed “oncosomes”) preisolated by differential centrifugation. Furthermore, the effect of resalting on the EM of sEVs and mEVs was evaluated. The EM of U87 sEVs and U87 mEVs showed an increase as the salt concentration decreased to 0.005% of the initial salt concentration. However, for the plasma sEVs and plasma mEVs, the electrophoretic mobility increased as the salt concentration decreased to 0.01% of the initial salt concentration and then increased to its initial value when the salt concentration decreased to 0.005% of the initial salt concentration. For both U87 and plasma sEVs and mEVs, the EM remained almost constant when the concentration of the particles changed and the salt concentration was kept the same as its initial value. This indicates that the EM of EVs is only a function of the salt concentration of the buffer and is independent of the concentration of the particles. The sEVs and mEVs were separated with cyclical ElFFF for the first time. The results indicate that ElFFF was able to fractionate the EVs, and a crescent-shaped trend was found for the retention time when the applied AC voltage was altered (increased).

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Section: Methodsmentioning

confidence: 99%

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confidence: 99%

Characterization of Human Glioblastoma versus Normal Plasma-Derived Extracellular Vesicles Preisolated by Differential Centrifugation Using Cyclical Electrical Field-Flow Fractionation

et al. 2020

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“…One difficulty in using El-FFF for biological vesicle separation is that salt concentration within typical physiological vesicle buffers is high enough to reduce the strength of the effective electric field within the El-FFF channel. 28 Salt must be removed via buffer substitution if effective vesicle separation using El-FFF is to be achieved. Conceivably, buffer substitution may influence El-FFF dependent exosome subfractionation.…”

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confidence: 99%

Exosome Isolation: Cyclical Electrical Field Flow Fractionation in Low-Ionic-Strength Fluids

et al. 2018

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The influence of buffer substitution and dilution effects on exosome size and electrophoretic mobility were shown for the first time. Cyclical electrical field flow fractionation (Cy-El-FFF) in various substituted fluids was applied to exosomes and other particles. Tested carrier fluids of deionized (DI) water, 1× phosphate buffered saline (PBS), 0.308 M trehalose, and 2% isopropyl alcohol (IPA) influenced Cy-El-FFF-mediated isolation of A375 melanoma exosomes. All fractograms revealed a crescent-shaped trend in retention times with increasing voltage with the maximum retention time at ∼1.3 V AC. A375 melanoma exosome recovery was approximately 70–80% after each buffer substitution, and recovery was independent of whether the sample was substituted into 1× PBS or DI water. Exosome dilution in deionized water produced a U-shaped dependence on electrophoretic mobility. The effect of dilution using 1× PBS buffer revealed a very gradual change in electrophoretic mobility of exosomes from ∼−1.6 to −0.1 μm cm/s V, as exosome concentration was decreased. This differed from the use of DI water, where a large change from ∼−5.5 to −0.1 μm cm/s V over the same dilution range was observed. Fractograms of separated A375 melanoma exosomes in two substituted low-ionic-strength buffers were compared with synthetic particle fractograms. Overall, the ability of Cy-El-FFF to separate exosomes based on their size and charge is a highly promising, label-free approach to initially catalogue and purify exosome subtypes for biobanking as well as to enable further exosome subtype interrogations.

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“…251−254 However, separation performance in constant voltage operation was limited due to the induced polarized layer on the surface of electrodes which weakened the effective electric field. 242,249,255,256 To increase the effective electric field, a pulse reversing technique was introduced as an alternative approach to discharge the polarized layer, thereby improving separation performance. 242,249,257−259 Using the pulsed voltage strategy, it is possible to optimize the retention behavior of nanoparticles via parameters such as pulse frequency, duty cycle, and waveform.…”

Section: Label-free Microfluidic Methods For Exosome Isolationmentioning

confidence: 99%

“…They showed that appropriate voltage waveforms suppressed the high diffusion rate of gold nanoparticles with sizes <50 nm and fractionated them with the highest resolution at a duty cycle of 75%. 242,257 Although electrical-FFF exhibits a significant capability to separate nanoparticles, it has rarely been tested for EV isolation. EVs are known to possess negative surface charge, thereby electrical-FFF could serve to gently separate EV subtypes based upon their electrophoretic mobility.…”

Section: Label-free Microfluidic Methods For Exosome Isolationmentioning

confidence: 99%

Label-Free Isolation of Exosomes Using Microfluidic Technologies

2021

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Exosomes are cell-derived structures packaged with lipids, proteins, and nucleic acids. They exist in diverse bodily fluids and are involved in physiological and pathological processes. Although their potential for clinical application as diagnostic and therapeutic tools has been revealed, a huge bottleneck impeding the development of applications in the rapidly burgeoning field of exosome research is an inability to efficiently isolate pure exosomes from other unwanted components present in bodily fluids. To date, several approaches have been proposed and investigated for exosome separation, with the leading candidate being microfluidic technology due to its relative simplicity, costeffectiveness, precise and fast processing at the microscale, and amenability to automation. Notably, avoiding the need for exosome labeling represents a significant advance in terms of process simplicity, time, and cost as well as protecting the biological activities of exosomes. Despite the exciting progress in microfluidic strategies for exosome isolation and the countless benefits of label-free approaches for clinical applications, current microfluidic platforms for isolation of exosomes are still facing a series of problems and challenges that prevent their use for clinical sample processing. This review focuses on the recent microfluidic platforms developed for labelfree isolation of exosomes including those based on sieving, deterministic lateral displacement, field flow, and pinched flow fractionation as well as viscoelastic, acoustic, inertial, electrical, and centrifugal forces. Further, we discuss advantages and disadvantages of these strategies with highlights of current challenges and outlook of label-free microfluidics toward the clinical utility of exosomes.

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Biased cyclical electrical field-flow fractionation for separation of submicron particles

Cited by 11 publications

References 14 publications

Characterization of Human Glioblastoma versus Normal Plasma-Derived Extracellular Vesicles Preisolated by Differential Centrifugation Using Cyclical Electrical Field-Flow Fractionation

Characterization of Human Glioblastoma versus Normal Plasma-Derived Extracellular Vesicles Preisolated by Differential Centrifugation Using Cyclical Electrical Field-Flow Fractionation

Exosome Isolation: Cyclical Electrical Field Flow Fractionation in Low-Ionic-Strength Fluids

Label-Free Isolation of Exosomes Using Microfluidic Technologies

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