Comparison of the capillary-channeled polymer (C-CP) fiber spin-down tip approach to traditional methods for the isolation of extracellular vesicles from human urine

Jackson, Kaylan K.; Powell, Rhonda R.; Marcus, R. Kenneth; Bruce, Terri F.

doi:10.1007/s00216-022-04023-5

Cited by 5 publications

(19 citation statements)

References 62 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In those latter data, the variability observed (<10% RSD) is due to the low protein values via the Bradford assay. Overall, these findings are in accordance with previous demonstrations of EV isolations using the C-CP tip, where the purity of the tip-recovered vesicles well exceeds the purity of vesicles processed using competitive ultracentrifugation or polymeric precipitation methods for EV isolation [35,40], on shorter time scales, low sample volumes, and lower capital costs.…”

Section: Protein Concentration Of Cell Culture Milieu and Purity Asse...supporting

confidence: 91%

“…The eluted EVs presented average diameters of 145–411 nm across the CCM sample collections across the incubation period, with an average diameter of 249 nm overall. In comparison to previously obtained populations of EVs collected using the C‐CP tip, the average diameter of the vesicles is 50–100‐nm larger than those obtained from human biofluids using the ACN or glycerol solvent systems and NTA determinations [35, 40]. The significant difference in EV size could potentially be due to the use of the Tween‐20 EV elution buffer or could be a basic characteristic of the suspension‐adapted HEK293 source.…”

Section: Resultsmentioning

confidence: 85%

“…The most common metric used to assess the purity of EV isolates is the fraction relationship between the number of EVs per mass of protein in the isolate, with >3 × 10 10 EVs µg −1 of protein considered to be “high purity” [47]. As recently demonstrated for the case of human urine‐derived EVs, this is one of the metrics where the C‐CP fiber tip method excels in comparison to other methods [40]. The relationship between EV and the protein concentration (i.e., purity) is depicted across the culture cycle in Figure 6B.…”

Section: Resultsmentioning

confidence: 99%

“…C-CP fiber micropipette tips were prepared through the previously described process [35][36][37]40]. The fibers were formed via melt-extrusion from bulk polyester (polyethylene terephthalate, PET) in the Clemson University School of Materials Science, having the form of an eight-pronged shape of ∼24 × 38-µm cross section.…”

Section: C-cp Spe Tip Assemblymentioning

confidence: 99%

“…The C-CP tip method allows for the isolation of highly concentrated EV samples in a quantitative and reproducible manner, using minute (100 µl) sample volumes [35][36][37]40]. These qualities are ideal in the case of small population (analytical) sampling of large-scale cell culture conditions to monitor the health of the cell line based on EV production.…”

Section: Quantification Of Recovered Evs As a Function Of Culture Tim...mentioning

confidence: 99%

See 4 more Smart Citations

Rapid isolation and quantification of extracellular vesicles from suspension‐adapted human embryonic kidney cells using capillary‐channeled polymer fiber spin‐down tips

Jackson

Marcus

2022

Electrophoresis

Self Cite

View full text Add to dashboard Cite

Exosomes, a subset of extracellular vesicles (EVs,, serve as biomolecular snapshots of their cell of origin and vehicles for intercellular communication, playing roles in biological processes, including homeostasis maintenance and immune modulation. The large-scale processing of exosomes for use as therapeutic vectors has been proposed, but these applications are limited by impure, low-yield recoveries from cell culture milieu (CCM). Current isolation methods are also limited by tedious and laborious workflows, especially toward an isolation of EVs from CCM for therapeutic applications. Employed is a rapid (<10 min) EV isolation method on a capillary-channeled polymer fiber spin-down tip format. EVs are isolated from the CCM of suspension-adapted human embryonic kidney cells (HEK293), one of the candidate cell lines for commercial EV production. This batch solid-phase extraction technique allows 10 12 EVs to be obtained from only 100-µl aliquots of milieu, processed using a benchtop centrifuge. The tip-isolated EVs were characterized using transmission electron microscopy, multi-angle light scattering, absorbance quantification, an enzyme-linked immunosorbent assay to tetraspanin marker proteins, and a protein purity assay. It is believed that the demonstrated approach has immediate relevance in research and analytical laboratories, with opportunities for production-level scale-up projected.

show abstract

Section: Protein Concentration Of Cell Culture Milieu and Purity Asse...supporting

confidence: 91%

Section: Resultsmentioning

confidence: 85%

Section: Resultsmentioning

confidence: 99%

Section: C-cp Spe Tip Assemblymentioning

confidence: 99%

Section: Quantification Of Recovered Evs As a Function Of Culture Tim...mentioning

confidence: 99%

See 3 more Smart Citations

Rapid isolation and quantification of extracellular vesicles from suspension‐adapted human embryonic kidney cells using capillary‐channeled polymer fiber spin‐down tips

Jackson

Marcus

2022

Electrophoresis

Self Cite

View full text Add to dashboard Cite

show abstract

In-line coupling of capillary-channeled polymer fiber columns with optical absorbance and multi-angle light scattering detection for the isolation and characterization of exosomes

Wysor,

Marcus

2024

Anal Bioanal Chem

Self Cite

View full text Add to dashboard Cite

Extracellular vesicles (EVs) have garnered much interest due to their fundamental role in intracellular communication and their potential utility in clinical diagnostics and as biotherapeutic vectors. Of particular relevance is the subset of EVs referred to as exosomes, ranging in size from 30 to 150 nm, which contain incredible amounts of information about their cell of origin, which can be used to track the progress of disease. As a complementary action, exosomes can be engineered with therapeutic cargo to selectively target diseases. At present, the lack of highly efficient methods of isolation/purification of exosomes from diverse biofluids, plants, and cell cultures is a major bottleneck in the fundamental biochemistry, clinical analysis, and therapeutic applications. Equally impactful, the lack of effective in-line means of detection/characterization of isolate populations, including concentration and sizing, is limiting in the applications. The method presented here couples hydrophobic interaction chromatography (HIC) performed on polyester capillary-channeled polymer (C-CP) fiber columns followed by in-line optical absorbance and multi-angle light scattering (MALS) detection for the isolation and characterization of EVs, in this case present in the supernatant of Chinese hamster ovary (CHO) cell cultures. Excellent correlation was observed between the determined particle concentrations for the two detection methods. C-CP fiber columns provide a low-cost platform (< $5 per column) for the isolation of exosomes in a 15-min workflow, with complementary absorbance and MALS detection providing very high-quality particle concentration and sizing information.

show abstract

Isolation and quantification of human urinary exosomes using a Tween-20 elution solvent from polyester, capillary-channeled polymer fiber columns

Bin Islam,

Marcus

2024

Analytica Chimica Acta

View full text Add to dashboard Cite

Comparison of the capillary-channeled polymer (C-CP) fiber spin-down tip approach to traditional methods for the isolation of extracellular vesicles from human urine

Cited by 5 publications

References 62 publications

Rapid isolation and quantification of extracellular vesicles from suspension‐adapted human embryonic kidney cells using capillary‐channeled polymer fiber spin‐down tips

Rapid isolation and quantification of extracellular vesicles from suspension‐adapted human embryonic kidney cells using capillary‐channeled polymer fiber spin‐down tips

In-line coupling of capillary-channeled polymer fiber columns with optical absorbance and multi-angle light scattering detection for the isolation and characterization of exosomes

Isolation and quantification of human urinary exosomes using a Tween-20 elution solvent from polyester, capillary-channeled polymer fiber columns

Contact Info

Product

Resources

About