Journal: Separation and Purification Technology

Article Title: Economical large-scale purification of extracellular vesicles from urine

doi: 10.1016/j.seppur.2023.126155

Figure Lengend Snippet: Fig. 1. Plasma-derived extracellular vesicles contain high levels of contaminants. (a–f) EVs were purified from plasma by enrichment via exoEasy Maxi columns (EEM) and SEC (a,d) or UF and SEC (b,e) or SEC only (c, f). (a–c) Tables depicts EV concentration determined by Zetaview® TWIN measurement as particles per ml of starting material and median of size distribution as determined in three independent experiments. (d–f) Size-distribution determined by DynaPro NanoStar mea surement, depicted as mean of three independent experiments each, indicate size range in nm, mode (Mo) is displayed in nm. g–i: negative-staining electron micrographs with 200 nm scale bars. EVs were purified from plasma with EEM (g), EEM and SEC (h) or with SEC only (i).

Article Snippet: EV size distribution was assessed on a DynaPro NanoStar (Wyatt Technology, Santa Barbara, California, Model Number: WDPN-14; Firmware-Version: 2.3.6.4) using disposable 25 μl Cyclic Olefin Copolymer (COC) cuvettes from the same company.

Techniques: Clinical Proteomics, Derivative Assay, Purification, Concentration Assay, Negative Staining

Journal: Separation and Purification Technology

Article Title: Economical large-scale purification of extracellular vesicles from urine

doi: 10.1016/j.seppur.2023.126155

Figure Lengend Snippet: Fig. 3. Evaluation of urine as a source for EV bulk purification using conven tional protocols. (a-c) EVs were purified from urine by salt precipitation. (d) EVs were purified from urine by PEG-precipitation (P) followed by a filtrating step (F) and heparin affinity purification. L = flow-through loading; W = flow- through washing; 1 M NaCl and 2 M NaCl: eluate. (a, d) EV concentration determined by Zetaview® TWIN measurement. Tables depict particles per ml of starting urine and median of size distribution as determined in three indepen dent experiments. (b) Size-distribution after 1st step of salt precipitation determined by DynaPro NanoStar measurement, depicted as mean of six in dependent experiments each, indicate size range in nm, mode (Mo) is displayed in nm. (c) Negative-staining electron micrograph of purified EV samples after 1st step of salt precipitation. Scale bars 200 nm.

Article Snippet: EV size distribution was assessed on a DynaPro NanoStar (Wyatt Technology, Santa Barbara, California, Model Number: WDPN-14; Firmware-Version: 2.3.6.4) using disposable 25 μl Cyclic Olefin Copolymer (COC) cuvettes from the same company.

Techniques: Purification, Affinity Purification, Concentration Assay, Negative Staining

Journal: Separation and Purification Technology

Article Title: Economical large-scale purification of extracellular vesicles from urine

doi: 10.1016/j.seppur.2023.126155

Figure Lengend Snippet: Fig. 5. Sequential purification steps enrich EV-sized particles from large volumes of urine. EVs were purified from urine by PEG-precipitation (P) followed by a filtration step (F), enrichment via exoEasy Maxi columns (EEM) (a, b) or ultrafiltration (UF) (c, d) and Size-exclusion chromatography (SEC). (e) EVs were purified from Huh7-cell conditioned medium. (a, c, e) EV concentration determined by Zetaview® TWIN measurement. Tables depict particles per ml of starting urine and median of size distribution as determined in twelve (a,c) or three (e) independent experiments. (b, d) Size-distribution determined by DynaPro NanoStar mea surement, depicted as mean of three independent experiments each, indicate size range in nm, mode (Mo) is displayed in nm.

Article Snippet: EV size distribution was assessed on a DynaPro NanoStar (Wyatt Technology, Santa Barbara, California, Model Number: WDPN-14; Firmware-Version: 2.3.6.4) using disposable 25 μl Cyclic Olefin Copolymer (COC) cuvettes from the same company.

Techniques: Purification, Filtration, Size-exclusion Chromatography, Concentration Assay