Hi, I want to load TMZ drug into extracellular vesicles (Exosomes) using incubation and sonication. I'm still stuck doing the incubation one. For the method, I mixed the TMZ and exosomes in the 1.5 microcentrifuge or 48-well plates. Then, I incubated the TMZ-Exos solution at room temperature for about 18 hours on the vortex mixer. After that, I washed the solution using ultracentrifuge at 100,000 g for 70 mins to isolate TMZ-loaded exosomes and non-loaded TMZ. Then, I resuspended the pellet (TMZ-loaded exosomes) and supernatant (non-loaded TMZ) in distilled water/PBS and read the absorbances using microplate reader to determine the concentration, as well as calculate the drug loading efficiency using standard curve. So, is there anything that needs to be improved or changed?

Also, I'm trying to add Tween-20 too to facilitate the drug loading into exosomes. Is it possible? Because I have read other papers and a few of them used Tween-20 to load drug into exosome.