Extracellular Vesicles
BioNavis Multi-Parametric Surface Plasmon Resonance (MP-SPR) instrument can measure EV size and concentration, cellular uptake, corona formation, affinity, and kinetics—all with a single tool!
Thorough Characterization of Extracellular Vesicles
Extracellular vesicles (EVs) are extensively studied for their unique characteristics and hold significant potential in various fields such as cancer research and therapy, diagnostic, regenerative medicine, and drug delivery. Characterization and interaction measurements of EVs are crucial parts of the research process.
Extracellular vesicle size and concentration
MP-SPR is an ideal tool for measuring EV size, especially for EVs smaller than 100nm due to high sensitivity of the technology. The unique multi-wavelength setup of MP-SPR, along with complete SPR curve measurements, are crucial features for size and concentration measurements.
BioNavis EV characterization kit provides straight forward approach for size and concentration characterization. The sensors can be functionalized by the user, the detection antibodies can be used depending on the EVs under investigation. Detection is not limited by the media used; hence, measurements can be performed also in complex media.
Extracellular Vesicle corona
BioNavis MP-SPR measurements can be performed with complex media such as cell culture media, plasma or serum. Plasma proteins have been shown to interact with EVs, and MP-SPR has been used to study the interaction between vesicles and serum proteins forming protein corona. As the measurements are performed on dynamic flow, this allows the study of proteins forming hard and soft corona on EVs, samples can be obtained for further analysis.
Affinity and kinetics
MP-SPR measures the binding affinity of EVs with high precision and their diagnostic molecules (such as antibodies, peptides, or aptamers) without labels. The dynamic flow principle also provides binding kinetics. The derived kinetic measurements can be used to improve the diagnostic molecule interaction parameters.
Cellular uptake
MP-SPR allows you to either form a lipid bilayer on the SiO2 sensor surface to mimic the cellular interaction or culture adherent cells on suitable sensor surface. MP-SPR’s flexible sensor slide design allows the use of standard cell culture protocols to form a confluent cell layer on a sensor slide ex situ.
Dynamic measurements can be used to observe the uptake of extracellular vesicles in living cells. This unique capability, enabled by the complete SPR curve measurement of MP-SPR, provides new insights into EV uptake. Additionally, after real-time MP-SPR measurements, the sensor slide can be further validated ex situ with microscopy techniques, including AFM and SEM.
Discover the world of MP-SPR
Selected MP-SPR Application Notes
Extracellular Vesicles: Frequently Asked Questions and Answers
-
How can you measure the size of extracellular vesicles (EVs)?
There are many techniques available, but MP-SPR measurements with two wavelengths enable accurate characterization of small extracellular vesicles size and concentration.
-
What are the most reliable methods for determining extracellular vesicle concentration in biological samples?
Many techniques require purification of the samples before measuring as presence of aggregates or proteins can lead to false positive results. MP-SPR is excellent technique for the crude sample measurement, thus, reducing the pre-purification of extracellular vesicles.
-
What is protein corona of extracellular vesicles?
Protein corona forms when extracellular vesicles (EVs) interact with biological fluid containing proteins (like plasma or serum), this interaction can alter the surface properties of the EVs, thus, impeding the intended interaction with cell or receptor.
-
How is the protein corona of extracellular vesicles studied?
MP-SPR, in real-time, can measure the formation of extracellular vesicle (EV) corona and also calculate the thickness of the protein corona on EV surface.
-
What techniques are available for analyzing EV uptake by living cells?
Most microscopy-based techniques (confocal or FLIM) require EVs to be prelabeled with lipophilic fluorescent dyes. These dyes have shown to either have form self-aggregates or alter the EV surface preventing interaction with cells. MP-SPR is a label-free technique as does not require any fluorophores allowing to study extracellular vesicle (EVs) and cell interaction in near native state.
-
How can the binding affinity and kinetics of extracellular vesicles with diagnostic molecules be measured?
An ideal diagnostic marker (molecule) should exhibit high sensitivity and selectivity. The flow-based system of MP-SPR instruments enables the measurement of binding affinity and kinetics with high sensitivity, making them extensively used for developing diagnostic markers.
-
How can extracellular vesicle–membrane interactions be studied?
Lipid bilayers can be pre-functionalized with selective receptors, simulating the cellular lipid bilayer while focusing on the interaction between extracellular vesicles (EVs) and these receptors. MP-SPR instruments, along with SiO₂ sensors, enable the preparation of lipid bilayers, and their unique fluidics allow the controlled introduction of EVs in a dose-dependent manner.
-
What methods are used to observe extracellular vesicles interactions in real-time?
Measurements carried out with dynamic MP-SPR instruments are performed under flow conditions, which can mimic shear forces similar to those in the body. The data is acquired in real-time and in a label-free manner, allowing for the elucidation of kinetic parameters.