An exosome (EV) derived from human plasma was analyzed using UB-100, an aqueous SEC (GFC) column for bionanoproduct analysis and preparative use. A combination of various detectors was use for the analysis.
UV (280 nm) detects general biologically derived impurities, fluorescence (FL; Ex 280 nm / Em 348 nm) primarily targets proteins (using Trp residues as indicators), and MALS light scattering (LS) provides a highly sensitive response particularly to large nanoparticles. Additionally, MALS offers an estimated RMS radius of the target.
Based on the MALS results, the fraction around 13 minutes is estimated to be the primarily EV components, and RMS radius indicates that it has a molecular weight distribution. Even though the EV sample derived from human plasma used in this analysis is of purified grade, peaks other than the EV fraction were observed in the UV and FL profiles, suggesting that a significant number of impurities is still present.
As seen, the UB series particle pores are designed to separate and purify not only profiles of the target bionano-range products (50 nm and above) including EV, but also separates them from biologically derived impurities. which is crucial for product’s quality evaluations.
Additionally, UB-100 employs 27 µm monodisperse particles, enabling low-pressure operation that accommodates workflows under high biological load and their use in lab-scale fractionation. Furthermore, its high alkaline durability supports alkaline solution cleaning, which is essential for high reproducibility and reduction of carryover. *Please Note: sales of UB columns will being in Jan 2025*
When trace and more precise analysis is the primary objective, please refer to OHpak SB-806 HQ, aqueous SEC (GFC) column, application.
Sample: 15 μL
Exosome from human plasma purification grade
Column：Shodex UB-100 (8.0 mm I.D. x 300 mm) Eluent：PBS (-) Flow rate：0.6 mL/min Detector：UV (280 nm) (small cell volume), Fluorescence (Ex. 280 nm, Em. 348 nm), MALS (Multi angle ight scattering) Column temp.：Room temp.