Rapid isolation of extracellular vesicles from diverse biofluid matricesviacapillary-channeled polymer fiber solid-phase extraction micropipette tips

Abstract

C-CP fiber spin-down tips are employed on a table-top centrifuge to isolate extracellular vesicles from diverse biofluids in less than 15 min. The EVs are isolated in high yields and purity, with the recoveries verified across multiple methods.

Facile, generic capture and on-fiber differentiation of exosomesviaconfocal immunofluorescence microscopy using a capillary-channeled polymer fiber solid-phase extraction tip

“…1. The isolation method, to the point of presenting purified EVs on the fiber surface, was executed as described previously, 23,24 with the initial labeling and imaging procedures following what is normal in the fluorescence immunoimaging field. 31 Following the basic demonstration of the methodology, efforts towards more rapid on-fiber processing were initiated.…”

“…That said, multiple previous efforts have demonstrated the ability to employ simple optical absorbance detection to quantify the total EV content from very diverse biological and culture media. [20][21][22][23][24]26 Those methods have been validated through use of diverse quantification modes (e.g. direct response functions and standard addition) as well as the common standard, nanoparticle tracking analysis (NTA).…”

“…The method's efficiency has been demonstrated in protein and exosome separations on C-CP fiber columns via HPLC [20][21][22] and by SPE on spin-down tips. 23,24 Overall, the process has proven to provide high EV yields (up to 1 × 10 12 EVs per mL) and extremely low concomitant protein content (<0.5% lipoprotein carryover in serum), 25,26 on time scales of less than 10 min, without any other form of pre-isolation. While no direct comparisons have been made, the SPE tips have yielded total exosome binding capacities of ∼8 × 10 11 EVs from commercial standards, 23 while the 30 cm columns more fitting for downstream processing have shown capacities ~5 × 10 12 for the far more complex, direct cell milieu purification.…”

Improving Isolation of Extracellular Vesicles by Utilizing Nanomaterials

“…Compared to the organic solvents employed in the reverse phase, salt containing buffer is much better for maintaining the biological activity of EVs. Taking advantage of this, C-CP fibers have expanded their application in solid-phase extraction workflows, rather than HPLC processing platforms, by being packed into a spin-down column or micropipette tip [77][78][79][80][81].…”

Advances in the Field of Micro- and Nanotechnologies Applied to Extracellular Vesicle Research: Take-Home Message from ISEV2021

“…Looking for a very fast (<15 min), easy to use and economic (<1 $/each) isolation method, Dr. Jackson (Clemson University, Clemson, SC, USA) presented capillarychanneled polymer (C-CP) fiber micropipette tips employed in a solid-phase extraction workflow, which allowed processing using low sample amounts (OD05.05). The method was proven to be suitable for clinical applications in terms of sample volumes and time scales, and also to be extremely versatile as it was tested for the EV isolation from human urine, saliva, cervical mucus, serum and goat milk matrices [28]. Another purification assay was designed for the high-throughput analysis of low abundance miRNAs of EVs and the evaluation of EVs' functional characteristics.…”