SEC/MALS Analysis of Exosome (2) (UB-100)

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.

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

  1.  
Chromtogram of Exosome
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 light scattering)
Column temp.
Room temp.

Sample Name Index

Operation Manual / Certificate of Analysis

Operation Manuals and Certificate of Analysis / Inspection Certificate for the following products can be downloaded here.

Product Name Index

Applications

Applications (Related Information)