Preparation of nanogel-immobilized porous gel beads for affinity separation of proteins: fusion of nano and micro gel materials

Abstract

We describe the preparation and evaluation of nanogel-immobilized porous gel beads (GB) for application as a protein purification medium. Nanogel particles (NP) that bind with the Fc fragment of immunoglobulin G (IgG) were immobilized on the pore surface of macroporous hard GB containing quaternary ammonium cations on the surface via multipoint electrostatic interactions. The amount of NPs that were irreversibly immobilized in 1 ml of GB slurry was determined to be~30 mg using fluorescent-labeled NPs. Images obtained via scanning electron microscopy established that the NPs were uniformly immobilized on the surface of the pores without blocking the macropores. The model target protein (IgG) was reversibly captured by the NPimmobilized GBs through NP-IgG interactions. NP-immobilized GBs have potential applications as novel affinity purification media for proteins, combining inexpensive and stable ligands with high-performance supports.

Effect of Chemical Crosslinking on Properties of Polymer Microbeads: A Review

“…Over the last two decades, natural and synthetic polymers were potentially used for various applications [19][20][21]. Both types of polymers have their own merits and demerits.…”

Matrices and Affinity Ligands for Antibody Purification and Corresponding Applications in Radiotherapy

“…Polymer nanoparticles (NPs) can also act as the affinity ligands of antibodies [ 97 , 98 , 99 ]. For example, the polymer hydrogel NPs (50–65 nm) showed high affinity for IgG, while little affinity for other proteins [ 97 ].…”

“…Their binding domain on IgG overlapped with that of protein A. In addition, synthetic nanogel particles that bound to the IgG-Fc fragment could be immobilized on a matrix to reversibly capture IgG [ 98 ]. The inexpensive and stable polymer NPs with the capacity of selectively binding to antibodies are significant alternatives to natural protein ligands for applications in antibody affinity purification.…”

High Reusability of Catalytically Active Gold Nanoparticles Immobilized in Core–Shell Hydrogel Microspheres

“…Metallic nanoparticles represent fascinating aspects such as flexible assembly behavior, which is interesting in the context of materials science, the behavior of the individual particles, and their electronic, magnetic, and optical properties, as well as their applications in catalysis and biology. − To improve the colloidal stability and endow the metallic nanoparticles with other properties in aqueous solution, colloidal carriers such as polymeric microspheres are usually applied, − as they exhibit a large specific surface area, fast response, the potential to be used in mass production, and high diffusibility, as well as ease of recovery and handling. − Hydrogel microspheres (microgels) are one of the most extensively studied classes of carriers, and they exhibit a three-dimensional cross-linked structure that swells in water. − Hybrid microgels represent attractive prospective components for functional materials given that they combine the desirable features of water-swollen microgels (e.g., high colloidal stability and stimuli-responsive properties) with the attractive properties of metallic nanoparticles. − So far, a variety of metallic nanoparticles immobilized in microgels have been developed for applications in, for example, sensors, − bioimaging, , and catalysis. − …”