Get to Know the qEV Gen 2 Range, Your Go-To for EV Isolation

Extracellular Vesicles
About /

The Gen 2 qEV range provides a strong foundation for extracellular vesicle isolation, paving the way towards a future where EVs shape clinical decisions and health outcomes. Here, we share an overview and key resources related to qEV Gen 2 columns.

The qEV Gen 2 range is comprised of Izon’s most advanced size exclusion chromatography columns. To achieve the best possible results, qEV Gen 2 columns are made from a high-performance resin developed specifically for the isolation of extracellular vesicles (EVs). Spanning six column sizes, the complete range is comprised of columns optimised for different sample loading volumes. This diversity enables qEV Gen 2 columns to provide a strong foundation for EV isolation across diagnostics, therapeutics, and fundamental research.

Benefits of qEV Gen 2 columns

qEV Gen 2 columns harness size exclusion chromatography to separate EVs from soluble protein, and deliver a highly purified EV isolate in an efficient and standardised manner. Alongside the Automatic Fraction Collector (AFC), which adds key elements of automation and customisation to EV isolation, qEV columns have become widely adopted across the EV field as a rapid, reproducible and gentle approach to EV isolation (Figure 1). Together, the AFC and qEV columns comprise the qEV isolation platform, which is critically – and uniquely – a scalable separation method.

The benefits of size exclusion chromatography as a technique underlie the reasons why the use of size exclusion chromatography is on the rise for EV isolation. To learn more specifically about why qEV columns are a cut above the rest, watch or read about 4 Reasons to Use qEV Columns for Extracellular Vesicle Isolation.

Figure 1. Publications featuring qEV Columns from 2014 to 2022.

Meet the complete Gen 2 qEV range

Through the use of a high-performance resin unique to the Gen 2 range, Gen 2 columns remove ~99% of contaminating soluble protein to provide a highly purified EV-containing isolate.

The qEV Gen 2 range is available in six column sizes, which accommodate everything from precious portions of blood samples dedicated to EV-RNA analysis, through to large volumes of cell culture media. The dimensions and features of each column have been optimised to enable the isolation of EVs from different sample loading volumes. qEV columns are versatile; in addition to the common sample types of plasma, serum, and cell culture media, they are used to isolate EVs from a wide range of other biofluids including milk, cerebrospinal fluid, urine, and reproductive fluids.

Figure 2. The complete Gen 2 range. Each column is optimised to suit a different sample loading volume: qEVoriginal Gen 2 (500 µL), qEVsingle Gen 2 (150 µL), qEV1 (1 mL, unique to the Gen 2 range), qEV2 Gen 2 (2 mL), qEV10 Gen 2 (10 mL) and the qEV100 Gen 2 (100 mL).

Each qEV Gen 2 column is available in two ranges, each of which will be familiar to current qEV customers. While both deliver on purity, there are differences between the two ranges. Columns from the 35 nm Series provides a greater recovery of smaller EVs, yet have more protein overlap as a trade-off of increasing recovery at the smaller end of the scale. The 70 nm Series, on the other hand, favours a higher recovery of larger EVs, and so offers less protein contamination. Deciding which is right for you will be a balance based upon your own needs. To help with this choice, visit the qEV overview page or browse qEV Gen 2 columns directly on the store:

Got questions about qEV columns? Check out our page of FAQs or talk to one of our team members to figure out which column is right for you.

Improved separation means more flexibility and choice with your isolation

The improved separation of the Gen 2 range provides an improved level of control when deciding which purified collection volumes (PCVs) to collect. As shown in Figure 3, if you study your elution profile sufficiently (and/or follow the recommendations in the user manual for plasma samples) you can collect different portions of your EV-containing isolate to prioritise EV concentration, recovery, or purity. The default values (listed in Gen 2 user manuals) provide a balance between EV recovery and purity.

Figure 3. The enhanced resolution of Gen 2 columns provides the opportunity to customise your collected volumes in a way that prioritises EV concentration, EV recovery, or EV purity.

Discounts available for bulk orders

If you’re planning to scale up, it’s important to know that you can take advantage of discounts when making bulk purchases. For example, the qEVoriginal Gen 2 column can be purchased in discounted packs of 100, and discounted multi-packs of the AFC V2 are available:

To receive a quote, simply get in touch with one of our helpful team members and they’ll be happy to assist.

qEV Legacy range to be discontinued

The development of the Gen 2 range reflects Izon’s dedication to providing the best possible EV isolation solutions to facilitate the adoption of EV-based clinical applications. With the improved performance of Gen 2 over Legacy, as measured by better purity in the qEVsingle Gen 2, qEVoriginal (Figure 4), qEV1, qEV2 Gen 2, qEV10 Gen 2 and qEV100 Gen 2, there has been a rapid adoption of Gen 2 columns – highlighting the importance of sample purity for the field.

Gen 2 columns will supersede the original qEV range, which have been renamed as ‘Legacy’ columns. The qEV Legacy range will be discontinued at the end of 2023.

Figure 4. Compared to Legacy qEV columns, Gen 2 qEV columns remove more protein and subsequently increase the EV-to-protein ratio.

Top: Protein present in pooled volumes (2 mL) of a human plasma sample isolated with Izon’s Automatic Fraction Collector, shown as a percentage of the total loaded protein. Protein was measured by bicinchoninic acid assay. Data shown for the qEVoriginal Legacy column and the Gen 2 qEVoriginal column in the 70 nm Series and 35 nm Series (0.5 mL loading volume).

Bottom: Number of extracellular vesicles (EVs) per microgram of protein, as measured by Tunable Resistive Pulse Sensing (the Exoid) and bicinchoninic acid assay, respectively. Data are shown for a human plasma sample separated using the AFC and qEVoriginal columns (0.5 mL loading volume) in the 70 nm and 35 nm Series.

We appreciate that navigating a change to Gen 2 may come with challenges; a new resin means a different isolation protocol will be required, and there’s a good chance you will need to re-optimise analysis downstream.

Read more about why results may differ when changing to qEV Gen 2 columns.

If you need support changing your protocols from qEV Legacy columns to qEV Gen 2 columns, please reach out and one of our support specialists will be able to assist.

As laid out in Table 1, the Buffer Volumes and Purified Collection Volumes differ between Gen 2 and Legacy columns, so it’s important to make sure you’re following the correct protocol in your new Gen 2 user manual. Otherwise, you may be missing a significant proportion of your EVs.

Table 1: Comparison of Buffer Volumes and Purified Collection Volumes Across Gen 2 and Legacy

*Note that the values referred to here use a different terminology between Gen and Legacy; elution profiles from Gen 2 columns (human plasma) have been characterised to enable the identification of a default buffer volume, which provides a balance of EV purity and recovery. This default buffer volume differs from the ‘buffer volume’ which is specified separately. Default buffer volumes and default purified collection volumes are programmed onto the Automatic Fraction Collector (AFC). When working with Gen 2 columns, refer to the table in your respective qEV user manual which specifies recommended parameters for different optimisation scenarios. For Legacy qEV columns, the buffer volume and PCV is listed in the respective user manual.

Due to its size, the qEV100 is not compatible with the AFC; refer to your respective qEV100 user manual for guidance on recommended parameters.

Struggling with Western blot? Start here.

qEV isolation platform a strong foundation for EV research

Gen 2 qEV columns, together with the Automatic Fraction Collector, provide a scalable and reproducible way forward for EV isolation. The enhanced separation of Gen 2 columns, as compared to the Legacy range, provides not just better purity – but also more flexibility to prioritise different sample compositions (Figure 3). While the original qEV range was initially established to provide a higher level of sample purity for Tunable Resistive Pulse Sensing measurement, qEV columns are widely used across a wide range of applications and workflows. Furthermore, EVs isolated from qEV columns can be concentrated using the qEV Concentration Kit, in preparation for protein or RNA analysis. Meanwhile, the qEV RNA Extraction Kit can help you further streamline sample preparation for EV-RNA analysis.

Enquire to discuss customised solutions

Did you know we collaborate with customers looking for customisation? We have custom-designed columns for unique sample types and also customise processes for the separation of large sample volumes (> 100 mL). If you’re not seeing exactly what you’re looking for, talk to us about our Customised EV Separation Solutions!

Get the latest on EV research

At Izon, we aim to share the most up-to-date information to help you navigate your EV research:

Plasma Extracellular Vesicles: Does Anticoagulant Type Matter?

Getting different results after changing isolation methods?

Browse a selection of recent qEV papers:

To get the latest, make sure you subscribe to our newsletter.


Related Articles

View All

Subscribe to our newsletter!

Get the latest qEV and TRPS news straight to your inbox.

Check out "Get to Know the qEV Gen 2 Range, Your Go-To for EV Isolation" on Izon

No items found.
No items found.
No items found.
No items found.
URL Copied!