Surface imprinting of pepsin via miniemulsion polymerization

Abstract

Surface imprinted polymers allow accessibility of the selective binding sites to large molecules such as proteins. In this work, small polymer particles offering a large surface area were prepared via miniemulsion polymerization in the presence of pepsin serving as a template molecule. The influence of four different functional monomers and of the amount of the template on the imprinting effect of pepsin was investigated. After the miniemulsion polymerization and a washing step, stable polymer suspensions with an average particle diameter of 400-600 nm and a specific surface area of 30-65 m 2 g À1 were obtained. The results of detailed rebinding experiments revealed that the highest imprinting effect was achieved with (3-acrylamidopropyl)trimethylammonium chloride as a functional monomer and a high amount of the template. These polymer particles also showed selectivity for pepsin against various proteins. This approach provides a fundamental step towards the development of synthetic protein receptors and protein scavenger materials useful in biomimetic assays and for clean-up in biotechnology.

Inhibitor-assisted synthesis of silica-core microbeads with pepsin-imprinted nanoshells

“…With applications in analytical chemistry, food analysis, medicine, and biotechnology, molecular imprinting techniques have been used in a variety of applications requiring synthetic recognition elements including solid-phase extraction, liquid chromatography, electrochromatography, drug delivery, and for biomimetic sensors. A method potentially circumnavigating this problem is surface grafting 8 , whereby micron-sized spherical beads serving as architectural core material are grafted with imprinted thin films. 6 However, imprinting of large molecules -and especially biomacromolecules such as proteins -remains a challenge due to their molecular weight and dimensions, limited stability in organic solvents, and thermodynamically caused structural flexibility in solution.…”

Binding performance of pepsin surface-imprinted polymer particles in protein mixtures

“…22 The water phase was prepared by dissolving pepsin (150 mg), APTMA (192 mg), and Lutensol AT50 (180 mg) in degassed water (15 g). 22 The water phase was prepared by dissolving pepsin (150 mg), APTMA (192 mg), and Lutensol AT50 (180 mg) in degassed water (15 g).…”

“…22 After polymerization, the water phase of the polymer suspensions was exchanged by centrifugation and redispersion cycles in order to remove pepsin, the surfactant, and any residual monomer. 22 After polymerization, the water phase of the polymer suspensions was exchanged by centrifugation and redispersion cycles in order to remove pepsin, the surfactant, and any residual monomer.…”

“…13 Efficient routes for obtaining such materials are core-shell emulsion 14 or miniemulsion polymerization techniques. 20 We have previously reported on the preparation of pepsin surface-imprinted polymer particles via miniemulsion polymerization, 22 whereby the influence of four different functional monomers, and the amount of the pepsin template on the imprinting effect were investigated. This onestep synthesis eliminates additional steps, as required for coreshell approaches.…”

“…22 Here, the maximum binding capacity and binding kinetics were investigated via batch rebinding experiments at varying initial pepsin concentrations, and at different time intervals. 22 Here, the maximum binding capacity and binding kinetics were investigated via batch rebinding experiments at varying initial pepsin concentrations, and at different time intervals.…”