Vesi-Bind Fc-mNG

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A new generation of engineered extracellular vesicles for functional studies and analytics

Vesi-Bind Fc products offer the unique capability of functionalising EVs with human IgG1, mouse IgG2 or rabbit IgG antibodies at their surface using their Fc domain. This new class biological nanoparticle has been developed by Samir El Andaloussi’s group at the Karolinska Institute, is an extracellular vesicle isolate from cells engineered to express the fluorescent protein mNeonGreen intraluminally in the presence of the transmembrane tetraspanin protein CD63, as well as a Fc-binding function at their surface. This construct allows the efficient binding of IgG antibodies via the Fc end, leaving the functional side accessible for binding experiments.

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Description

A new generation of engineered extracellular vesicles for functional studies and analytics

Vesi-Bind Fc products offer the unique capability of functionalising EVs with human IgG1, mouse IgG2 or rabbit IgG antibodies at their surface using their Fc domain. This new class biological nanoparticle has been developed by Samir El Andaloussi’s group at the Karolinska Institute, is an extracellular vesicle isolate from cells engineered to express the fluorescent protein mNeonGreen intraluminally in the presence of the transmembrane tetraspanin protein CD63, as well as a Fc-binding function at their surface. This construct allows the efficient binding of IgG antibodies via the Fc end, leaving the functional side accessible for binding experiments.

Vesi-Bind Fc-mNG enables and facilitates the following experiments:

  • Fast evaluation of the binding efficiency of antibody-modified EVs to recipient cells
  • Modelling for the development of LNP and nanomedicines candidates
  • Efficient co-localisation experiments on nanoparticle analysis and super resolution microscopy platforms
  • Positive control of (nano)flow cytometry measurements on two fluorescence channels

Fast functionalisation of the EV surface with monoclonal antibodies:
Vesi-Bind Fc are extracellular vesicles designed to bind the Fc portion of antibodies, allowing the variable regions to be displayed for antigen recognition, can be decorated with various types of antibodies. These Fc-binding EVs can thus be targeted to virtually any tissue (better: antigen) of interest.

Additionally, they display an intraluminal mNeonGreen fluorescent signal, which can be used as a reporter protein domain for uptake & functional assays.

Flexible EV co-localisation experiments
Co-localisation of unknown EV samples can be difficult to perform given the challenges of the preparation and immunolabeling steps. Vesi-Bind Fc-mNG offers a readily fluorescent source of extracellular vesicles fluorescent on the FITC channel (excitation 488nm, emission 525nm) as well as a second colour via functionalisation with a primary conjugated Fc antibody.

A reliable reference material for the standardisation of flow cytometry measurements
(Nano)flow cytometry is now extensively used for extracellular vesicle characterisation including the quantification of particles positive for the presence of specific biomarkers, including tetraspanin (CD9, CD63 and CD81). Assessing the sensitivity of analytical instruments for the detection of particles as small as EVs has always presented a challenge, because of their small optical radius and low density. Being an actual EV isolate, Vesi-Bind Fc-mNG offers the desired optical properties together with fluorescence on two or more channels, allowing the fine-tuning of powerful flow cytometers and sorter for EV work by serving as a new, versatile reference material for studying antibody binding to EVs.

Stable on the bench, as well as in the freezer
Exosomes are known to degrade rapidly once exposed to positive temperatures, hampering short-term projects, as well as analyses. As it is stabilised with Vesi-Safe buffer, Vesi-Bind Fc-mNG can be safely stored for prolonged periods at -80°Cto -20°C, as well as at operational temperatures during incubation and analysis, without observable degradation.

Additional information

Options

Vesi-Bind Fc-mNG only, Vesi-Bind Fc-mNG & Vesi-Ref CD63-mNG

Technical Description

Concentration (mNG+/mL, measured by IFCM)

MFI (FITC MESF) for unstained samples

 Estimate of % engineered (ratio of mNG+APC+ vs APC+)

Mode diameter NTA (Nanosight)

Mode diameter NTA (ZetaView)

Volume

Buffer

3.85 E9 particles/mL

815.6285

88.90%

128.7nm

126.1nm

 100µL

Vesi-Safe