Abstract
In this work, magnetic functionality was introduced to cross-linked acrylamide-based particles via the in situ coprecipitation of iron oxide nanoparticles within the hydrogel particle interior. Cibacron Blue F3G-A was then incorporated onto the magnetic hydrogel scaffold to facilitate the harvest of targeted protein species. The dye-loaded magnetic particles were physically characterized, and their protein sequestration performance was investigated. The results of these studies indicated that dye-loaded magnetic particles sequestered a greater amount of lower molecular weight proteins from the test solution than was achieved using reference particles, dye-loaded cross-linked N-isopropylacrylamide-based core-shell particles. This difference in protein harvesting ability may reflect the higher degree of dye-loading in the magnetic particles relative to the dye-loaded core-shell particles.
Highlights
Low-abundance, low-molecular weight peptides, proteins and other biomolecules, collectively known as biomarkers, can provide important information about the physiological state of an organism.By studying biomarkers, diseases such as cancer can be diagnosed in its early stages, thereby improving the overall prognosis for the patient
We report here the design, synthesis and characterization of hydrogel particles impregnated with magnetic iron oxide nanoparticles and baited with Cibacron Blue F3G-A
Hydrogel microspheres synthesized by precipitation polymerization typically have uniform size distributions and are free of contaminants
Summary
Low-abundance, low-molecular weight peptides, proteins and other biomolecules, collectively known as biomarkers, can provide important information about the physiological state of an organism. Cross-linked N-isopropylacrylamide (pNIPAm)-based hydrogel particles with incorporated affinity baits acrylic acid (AAc) and Cibacron Blue F3G-A have shown an ability to rapidly, in one step, sequester, concentrate and protect from proteolytic degradation low molecular weight proteins and peptides from serum, thereby allowing the analysis of low-abundance labile biomarkers [1,2]. The iron oxide shell may complicate the sequestration of low molecular weight species by hindering the target molecules from entering the porous hydrogel particle Another commonly used approach involves polymerizing a monomer mixture in the presence of surfactant-coated inorganic magnetic nanoparticles via emulsion and microemulsion polymerization. In order to affix the affinity bait, Cibacron Blue F3G-A, to the hydrogel scaffold, it was necessary to introduce suitable functionalizable groups into the polymer framework This was achieved by subjecting the magnetized acrylamide particles to controlled Hofmann degradation, which effectively converted a fraction of the acrylamide carboxamide groups to primary amines. The swelling and physical properties of the resulting baited magnetic particles were characterized, and their ability to effectively harvest and concentrate low abundance peptides and proteins while excluding larger higher molecular weight proteins was evaluated
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