Norovirus VLP derived from cell culture supernatant was analyzed using UB-50, 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 culture-derived impurities, meanwhile MALS light scattering (LS) provides a highly sensitive response particularly to large nanoparticles and estimated RMS radius of the target.
In this application, the results of transmission electron microscopy (TEM), which directly observes the VLP formation based on their shapes and sizes, was also compared. From the TEM imaging, formation of well-structured VLPs were observed. MALS result confirmed that the particle size corresponding to TEM image was observed around 16 minutes. Even though the norovirus VLP sample used in this analysis is of purified grade, RMS radius indicates the presence of larger particles (such as aggregates) compared to normal VLPs. Additionally, the UV profiling suggests that a significant number of biologically derived impurities were 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 below), but also separates them from biologically derived impurities. which is crucial for product’s quality evaluations.
Additionally, UB-50 employs 26 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-805 HQ, aqueous SEC (GFC) column, application.
Sample: 15uL
Norovirus VLP purification grade
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Column: Shodex UB-50 (8.0 mm I.D. x 300 mm) Eluent: PBS (-) Flow rate: 0.6 ml/min Detector: UV (280 nm) (small cell volume), MALS (Multi angle light scattering) Column temp.: Room temp.
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