One pot synthesis of surface PEGylated core-shell microparticles by suspension polymerization with surface enrichment of biotin/avidin conjugation

Core-shell microparticles that consist of poly(vinyl neodecanoate) (VND) crosslinked with poly(ethylene glycol dimethacrylate) (EGDMA) as the core and poly(ethylene glycol methacrylate) (PEGMA) (Mn = 360 or Mn = 526 g . mol(-1)) as the shell have been synthesized using suspension polymerization by a conventional free radical polymerization process. Interfacial tension and stability tests show that PEGMA acts as an amphiphilic macromonomer and is located on the oil/water interface of the suspension system, thus forming an outer layer during the polymerization. Kinetic studies of the monomers' conversion of VND, EGDMA, and PEGMA have been carried out using (1)H NMR spectroscopy. EGDMA and PEGMA were found to have faster reaction rates compared to VND. Moreover, scanning electron microscopy showed that the polymerization of these particles starts from the shell and finishes towards the core. Consequently, the resulting microsphere is found to have a multi-layer structure. Biotin was covalently bound to the surface by the PEGMA hydroxy groups. Conjugation of biotin with streptavidin PE (phycoerythrin) was subsequently carried out. Confocal microscopy was used to confirm the presence of fluorescing streptavidin. The amount of avidin conjugated to the microspheres was calculated by the release of a 2-(4-hydroxyphenylazo)benzoic acid/avidin complex using UV/vis spectroscopy. One avidin molecule was found to occupy 7 nm(2) on the surface of the microspheres.