I will start to fabricate micro/nanoparticles for growth factor release and I check out a procedure quite nice, however, it is lacking some details and since I don't have much knowledge doing this I also have some questions regarding the procedure. So any help is welcome.

This is the article "https://onlinelibrary.wiley.com/doi/abs/10.1002/bit.21822"

"We first prepared microspheres by the DE technique. A 250 mL solution of protein, containing CNTF and the excipient BSA (1:20 molar ratio of CNTF:BSA) in 1X PBS, was added to a solution of 200 mg PLGA in 7 mL dichloromethane (DCM). The resulting mixture was sonicated on ice for 10 s at 50% duty."

Is it possible maybe to create the emulsion by another method, maybe vortex ?

"The microspheres were centrifuged, washed three times with deionized water, freeze-dried, and stored over dessicant at -20C. "

After each step that I wash the particle I have to centrifuge, right? Otherwise, I will lose the particles. In this article, they don't describe the RPM/g? I saw some articles for like 1-3h and 1000-10.000 RPM. What would be acceptable?

"A 300 mL mixture of protein and surfactant (CNTF/BSA and AOT) in various concentrations and ratios were added to a solution consisting of 200 mg PLGA dissolved in a 5 mL mixture of DCM solvent and trifluoroethanol (TFE) cosolvent [1:4, DCM:TFE (v/v)]. "

Is it possible to substitute the AOT with another surfactant? Pluronic? I saw that is quite used for particle fabrication. Why in this case we have to use a combination of 2 different solvents?

Thanks

best regards