Separation of 100 nm-sized nanoparticles using a poly-Lys-modified monolith column

Area of Science:

• Analytical Chemistry
• Nanotechnology
• Biomedical Engineering

Background:

• Nanoparticles, including drug carriers and extracellular vesicles, are crucial in medicine but challenging to separate using conventional HPLC.
• Existing HPLC methods struggle with nanoparticle separation, leading to clogging, collapse, and low efficiency.

Purpose of the Study:

• To develop an efficient HPLC method for separating ~100 nm nanoparticles.
• To evaluate the performance of a poly-Lys-modified monolithic column for nanoparticle analysis.

Main Methods:

• Preparation of a poly-Lys-modified monolithic HPLC column.
• Analysis of anticancer drug-encapsulated nanoparticles (Doxil®) and small extracellular vesicles (sEVs).
• Utilized gradient elution with varying 2-amino-2-hydroxymethyl-1,3-propanediol concentrations.

Main Results:

• A high surface coverage poly-Lys column effectively retained nanoparticles via ion-exchange.
• Small extracellular vesicles (sEVs) with higher negative zeta potential (-45.5 V) showed stronger retention than Doxil® (-24.4 V).
• Successful separation of Doxil® and sEVs within 10 minutes using gradient elution.

Conclusions:

• The developed poly-Lys-modified monolithic column enables rapid and efficient separation of nanoparticles and sEVs.
• This method is valuable for assessing the safety and performance of nanoparticles in biomedical applications.
• The column offers a significant advancement over conventional HPLC for nanoparticle analysis.