A new age of EV concentration is here: the qEV Magnetic Concentration Kit allows you to concentrate EVs that have been purified using qEV columns. Using functionalised magnetic particles, you can concentrate EVs more quickly and cleanly than before – without the need for centrifugation or time-consuming protocols.
Developed as part of our expanding suite of new products for the EV diagnostic field as part of qEV PurePath for Diagnostics, the qEV Magnetic Concentration Kit unshackles you from centrifugation when concentrating EVs.
Why concentrate extracellular vesicles?
Isolating EVs from conditioned media and biofluids alike can leave you with a dilute sample, either because you had a dilute sample to begin with (e.g., conditioned media) or dilution occurred during the isolation workflow itself. qEV columns provide a purer EV isolate than competing methodologies (Figure 1A), making them the isolation method of choice for EV diagnostics.
Our analysis of the literature also shows that, contrary to popular belief, the use of size exclusion chromatography (SEC)-based qEV columns results in a higher yield than does ultracentrifugation, and no difference as compared to precipitation (Figure 1B). Regardless of your isolation methodology, further concentration will be needed for many applications.
What are the current options for concentrating EVs?
For large volumes, tangential flow filtration is ideal for concentrating EV isolates. When working with smaller volumes, such as those used in diagnostics, centrifugal concentration filters (e.g., Amicon Ultra) and capture particles (e.g., the qEV Concentration Kit) are two major options that are currently available.
However, both are time-consuming. The method involving centrifugal filters takes on average between 75 and 90 minutes to complete, according to the manufacturer’s protocol. Similarly, the protocol for our own qEV Concentration Kit (non-magnetic) takes about the same amount of time.
Whilst this time requirement may be okay for many lab settings, it will stifle productivity when isolating samples at scale – and may make diagnostic tests unworkable in the real world. Additionally, the need for a centrifuge for both methods limits automation, again impacting upon scalability. For these reasons, we have developed a new solution.
Introducing the qEV Magnetic Concentration Kit
The qEV Magnetic Concentration Kit is an all-in-one system for concentrating intact EVs isolated using qEV columns. To help facilitate EV concentration, the kit employs Magnetic Nanotrap® Extracellular Vesicle Particles produced for us by Ceres Nanosciences. Nanotrap® Particles are composed of hydrogel polymers that have been functionalised with chemical affinity baits. The nature of this product lends itself particularly well to automated isolation, allowing for maximal efficiency in scaled EV isolation for diagnostic purposes.
How long does it take to complete the qEV Magnetic Concentration Kit protocol?
After adding the specified volume of Magnetic Nanotrap® Particles to the EV isolate, samples are incubated at room temperature for 10 minutes (while being mixed with a tube roller or inverter). Next, a strong magnet (e.g., as part of a magnetic rack) is applied to pellet the EV-Magnetic Nanotrap® Capture Particle complexes for 2 minutes. With the magnet still applied, the supernatant can be removed. After removing the magnet, the pellet is resuspended in the desired buffer (e.g., a lysis buffer). Altogether, this procedure takes around 20 minutes to complete, making it around 4 times faster than other methods. As the kit does not require a centrifugation step, the protocol has the potential to be fully automated.
Just how well does the qEV Magnetic Concentration Kit work?
Given biomarker analysis is the most likely application of the qEV Magnetic Concentration Kit, we decided to see how much more easily an EV-associated miRNA could be detected following concentration. As you can see in Figure 3, the qEV Magnetic Concentration Kit resulted in a high level of concentration (p<0.05), which made the marker in question, miR-16, much more readily detectable than in unconcentrated samples. This increase in detectability could be the difference between a valuable EV biomarker being identified, or not.
The actual degree of concentration is variable, and is controlled by you. This is because the qEV Magnetic Concentration Kit leaves you with a pellet, and the volume in which the pellet is resuspended has a large bearing on the degree of concentration. For protein or RNA analysis, the most concentrated samples can be achieved by directly resuspending in the lysis buffer of choice, thereby preventing unwanted dilution.
What is the qEV Magnetic Concentration Kit suitable for?
The qEV Magnetic Concentration Kit enables you to capture and concentrate EVs before conducting downstream analyses that require highly enriched EV samples or small volumes. For example, concentrated samples are suitable for nucleic acid analysis (e.g., PCR or sequencing) and protein analysis (i.e., Western blots or mass spectrometry). Note that particle binding is irreversible, meaning that the qEV Magnetic Concentration Kit is not suitable for functional analysis or therapeutic applications.
Capture and concentrate intact EVs post qEV isolation
The qEV Magnetic Concentration Kit offers an excellent option for concentrating EVs following qEV isolation. The use of a magnet to pellet the EV-Magnetic Nanotrap® Particle complexes makes this method a clean and highly scalable and automatable option for the diagnostics field. It is also much faster than other methodologies, allowing you to process four samples in the time it would take you to process one with alternative methods. This makes the qEV Magnetic Concentration Kit the ideal partner for diagnostic applications, unshackling you from centrifugation in yet another step of your EV isolation workflow.
View on the online store: qEV Magnetic Concentration Kit
