Progressive Saturation Improves the Encapsulation of Functional Proteins in Nanoscale Polymer Vesicles

摘要: To develop a technique that maximizes the encapsulation of functional proteins within neutrally charged, fully PEGylated and nanoscale polymer vesicles (i.e., polymersomes). Three conventional vesicle formation methods were utilized for myoglobin (Mb) in polymersomes varying size, PEG length, membrane thickness. Mb concentrations monitored by UV–Vis spectroscopy, inductively coupled plasma optical emission spectroscopy (ICP-OES) bicinchoninic acid (BCA) assay. Suspensions subject to protease treatment differentiate amounts surface-associated vs. encapsulated Mb. Polymersome sizes morphologies dynamic light scattering (DLS) cryogenic transmission electron microscopy (cryo-TEM), respectively. Binding release oxygen measured using Hemeox analyzer. Using established “thin-film rehydration” “direct hydration” methods, was found be largely with negligible aqueous polymersome suspensions. Through iterative optimization, novel “progressive saturation” developed greatly increased final (from   2.0 mg/mL solution), weight ratio Mb-to-polymer could reproducibly obtained  4 w/w% Mb/polymer), as well overall efficiency  90%). Stable verified cryo-TEM; suspensions also displayed no signs aggregate for > 2 weeks assessed DLS. “Progressive further variety other proteins, ranging size from 17 450 kDa. Compared increases quantities may polymersomes.