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Application note:

Tangential Flow Filtration for efficient depletion of nanoparticles contained in Fetal Bovine Serum

Sirin Korulu Koc, Jevgenia Dõrina, Paolo Guazzi; HansabioMed Life Sciences

Fetal Bovine Serum (FBS) or Fetal Calf Serum (FCS) are commonly used supplements in cell culture media.
However, using FBS, nanoparticles of bovine origin, such as lipoproteins and EVs, are introduced into the
culture medium, where they not only can modulate the signaling of the recipient cells but also contaminate
the cell-derived vesicle preparation, co-isolating with them during the EV purification phase.
To date, several supplements or EV depleted sera are commercially available, as well as several protocols
have been developed to remove FBS nanoparticles, mainly based on ultrafiltration or ultracentrifugation
methods.
In this application note, we show how our TFF-EVs can be used to remove nanoparticles from FBS in one single
filtration shot, working directly under the safety cabinet. A simple, fast and inexpensive way to drastically
reduce the presence of nanoparticles in FBS, without impacting the cell growth and viability, and by preserving
the sterility.

HansaBioMed Life Sciences and Finnish Red Cross Blood Service start a join effort forHansaBioMed Life Sciences and Finnish Red Cross Blood Service start a join effort fordeveloping next generation of Extracellular Vesicles from donated blood as referencematerial for researchers.

The new cooperation is set under the heads of agreement and is aimed to offer highly characterized andstandardized EV material for life science market to further accelerate the development of the EV applications indiagnostics and therapy.

FLuoEVs: Purified EVs expressing Fluorescent proteins (lyophilized)

FLuoEVs are stably-fluorescent EVs expressing the fluorescent protein EGFP (green) BFP (blue) or mCherry (Red).

FLuoEVs are extracellular vesicles purified by combination of Tangential Flow Filtration (TFF) and Size Exclusion Chromatography (SEC) from cells engineered in order to express the fluorescent proteins as fusion protein with the tetraspanin CD9, CD63 or CD81.

Working with Plant EVs

Plant EVs, called also plant EV-like nanoparticles (ELNs), are attracting interest from researchers in academic sector, but also from the nutraceutical and the cosmetic industry. Indeed, plant EVs, such as the EV produced by mammalian cells, reflect the properties of the parental cells, can be purified from the plant leaves, stems, or juice, easily sterilized by filtration, and long-term stored. We started studying and working with extracellular vesicles from plant two years ago, in the context of a research project aimed to evaluate the particles produced by plants, in particular from Estonian origin, as potential source of regenerative and protective properties for skincare. In this period, we have developed expertise and methods for the purification and the characterization of vesicles from different plants, including rhizomes (ginger, ginseng), tubers (potato), Allioideae (garlic, onion), Cruciferae (cabbage family) and plant juices (coconut water, pineapple juice, orange and lemon juice). Our current workflow includes the preparation of the plant extract by plant juicer, the EV purification by Tangential Flow Filtration (TFF) or combination of TFF and Size Exclusion Chromatography. Among the particle analysis we can perform NTA, phenotyping by detecting the marker TET8, morphology analysis by Transmission Electron Microscopy.

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