Encapsulation of peroxidase by polymerizing acrylic acid monomers in "clean" polyelectrolyte microcapsules
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abstract
Encapsulation of macromolecules in microscale carriers has numerous applications in biotechnology, medicine, and chemical processing. Stability of encapsulated biomaterial is a significant draw-back especially for biosensor applications, which can be compensated by immobilization of a large quantity of biomaterial. A facile method for electrostatically-driven attraction and encapsulation of large quantities of macromolecules via in situ polymerization of charged monomers in polyelectrolyte microcapsules is described. The technique is based on fabrication of hollow microcapsules using the versatile Layer-by-Layer self assembly method, followed by loading and polymerization of acrylic acid monomeric units within the capsule cavity. Anionic acrylic acid monomers were polymerized by free-radical polymerization to form an anionic interior, and cationic horseradish peroxidase (HRP) was spontaneously loaded from the exterior solution. Using confocal microscopy, the loading was studied using fluorescent-labeled enzyme, from which it was observed that the protein was rapidly attracted through the capsule walls and stably entrapped in the interior. With an increase in the quantities of monomers and initiator, the quantity of HRP encapsulated and corresponding catalytic activity also increased. This methodology can be applied towards efficient and stable encapsulation of other charged macromolecules within hollow templates by selection of appropriate charged monomers with reactive groups that provide a stable environment to support the native function of the encapsulated species. Copyright 2007 American Scientific Publishers All rights reserved.
