Sonication is a suitable method for loading nanobody into glioblastoma small extracellular vesicles

Area of Science:

• Biotechnology
• Cancer Research
• Drug Delivery

Background:

• Glioblastoma necessitates advanced therapeutic strategies due to its high mortality rate.
• Nanobodies offer a promising avenue as nano-sized biotherapeutics.
• Efficient delivery systems are crucial for enhancing nanobody efficacy, especially for intracellular targets.

Purpose of the Study:

• To investigate small extracellular vesicles (sEVs) as a delivery system for anti-vimentin nanobody (Nb79).
• To evaluate different methods for loading Nb79 into sEVs.
• To assess the therapeutic effect of Nb79-loaded sEVs on glioblastoma cell survival.

Main Methods:

• Isolation of sEVs from glioblastoma cells via ultracentrifugation.
• Loading of Nb79 into sEVs using cell incubation, passive loading, or sonication.
• Confirmation of Nb79 loading using Western blot and electron microscopy.
• Assessment of sEVs' impact on glioblastoma cell viability using WST-1 assay.
• Nanoparticle tracking analysis for sEV size determination.

Main Results:

• Sonication proved to be a successful method for loading Nb79 into sEVs, confirmed by Western blot and electron microscopy.
• Cell incubation for Nb79 loading was unsuccessful and led to significant cell death.
• sEVs without Nb79 increased the survival of U251 and NCH644 cells by 20-25%.
• Nb79-loaded sEVs decreased the survival of NCH421k cells by 11%.

Conclusions:

• Sonication is an effective technique for loading nanobodies into extracellular vesicles.
• Nb79-loaded sEVs demonstrate potential in reducing glioblastoma cell survival.
• This approach offers a viable strategy for targeted delivery of protein-based therapeutics.