While qEV columns are synonymous with extracellular vesicle (EV) isolation, their use can be employed for a wider range of nanoparticles, including liposomes and LNPs.
Unfamiliar with these two nanoparticles? Here's a quick recap.
What are liposomes and lipid nanoparticles?
Liposomes are one of the earlier drug delivery tools, with their name deriving from the spherical lipid bilayer structure which resembles cell membranes. Since the inception of liposomes, strides in nanomedicine have led to the evolution of LNPs (Figure 1).1 The significance of LNPs skyrocketed with the development of the mRNA-based covid-19 vaccines, and their applications are likely only in their infancy.
The need for effective LNP isolation
As LNP-based treatments rise in prominence, so does the need for effective and reliable isolation methods. How LNPs are developed – from formulation to storage – impacts their therapeutic efficacy and safety. Ensuring that LNPs are free from contaminants, such as smaller particles and proteins, is crucial for standardising development and optimising therapeutic outcomes.
Multiple isolation methods exist – each with their unique advantages and limitations. Size exclusion chromatography (SEC), centrifugation, and filtration methods are available to purify LNPs.2,3 However, ultracentrifugation (UC) uses large centrifugal forces, which can be damaging to nanoparticle structural integrity.4 Additionally, filtration and centrifugation methods may not offer the same level of precision in removing smaller contaminants as SEC, as detailed in our article comparing all three isolation methods (Figure 2).
*References 1-6 from Beyond Ultracentrifugation: Embracing the qEV100 for Enhanced Large-Scale EV Isolation Purity and Efficiency
Size exclusion chromatography (SEC) is a widely used method for gently separating particles based on size, and are available as DIY or commercial kits. While DIY kits are often cheaper, they may lack the reliability that comes with standardised commercial qEV columns.
The benefits of the qEV isolation platform
Izon's qEV columns are a commercially available SEC system, specifically designed for nanoparticle isolation. They enable a standardised and gentle isolation process, that allows nanoparticles to maintain their integrity while being separated from smaller contaminating particles. In terms of LNPs, qEV columns prevent structural damage which is essential for preserving their therapeutic function.
Plus, qEV columns are available in a wide range of sizes, catering for fundamental research and diagnostic development, as well as large-volume biopharmaceutical production. From 150 µL to 100 mL sample loading volumes, our ''off-the-shelf'' qEV columns offer the ability to scale isolation efforts while maintaining consistency and reproducibility. If you're working with larger sample volumes, we also offer customised qEV columns to best suit your sample volume needs.
The size range of the particles you're wanting to isolate will determine which qEV resin series you should use. If you're looking to maximise LNP recovery from a sample, then the 20 nm series is your best bet. Have a look at our qEV series comparison webpage for a more detailed analysis between the three resin series.
Our range of qEV columns are compatible with a selection of automation instruments (Figure 3), allowing you to streamline and scale nanoparticle isolation – whether it be for fundamental, therapeutic or diagnostic research purposes.
The use of qEV columns for lipid-based nanoparticle purification
While LNPs may be most widely known for their involvement in the mRNA-based COVID-19 vaccine, their applications as biological carriers extend much further. They are used to deliver a range of therapeutic molecules, such as RNA, mRNA and siRNA, and are instrumental in gene editing and vaccine development.
qEV columns have previously been used to isolate lipid-based nanoparticles. In a study investigating shelf life of liposomes containing anti-inflammatory steroids, qEV columns helped assed the drug entrapment capacity of liposomes.5 By processing the supernatant through qEVoriginal 70 nm Gen 2 columns, researchers were able to remove the free dye, isolating the drug-loaded liposomes from the free drug based on size.
In an additional study, qEVoriginal columns were employed to purify antibody conjugated LNPs. These LNPs were then used to effectively deliver therapeutic RNA in vivo to haematopoietic stem and progenitor cells (HSPCs) in rodents.6
Effective and reproducible isolation essential to the future of LNP innovations
As nanomedicine continues to expand, effective and reliable LNP isolation methods will play a crucial role in developing advanced therapies. qEV columns offer a unique, scalable solution for LNP isolation – enabling researchers to develop more effective nanoparticle-based treatments. With growing applications in drug delivery, gene editing, and vaccine development, effective LNP isolation will be key to success of future biomedical innovations.
