Magnetic IDA‐modified hydrophilic methacrylate‐based polymer microspheres for IMAC protein separation

Abstract

Preparation of a new type of magnetic non-porous poly(2-hydroxyethyl methacrylate-co-glycidyl methacrylate) microspheres with hydrophilic properties containing coupled iminodiacetic acid (IDA) is described. The prepared microspheres were used for the immobilization of Ni(II) or Fe(III) ions to show their application in protein binding studies. Human IgG was bound to magnetic Ni(II)-IDA-modified microspheres and conditions of its adsorption and elution were optimized. Non-specific binding of the protein to magnetic microspheres in the absence of Ni(II) ions was low. Fe(III) ions immobilized on magnetic IDA-modified microspheres were used for the specific binding of porcine pepsin, as a model phosphoprotein. The ability of phosphate buffer to release the adsorbed enzyme from the microspheres and a low adsorption of the dephosphorylated protein indicate the participation of phosphate groups in the pepsin interaction. The elaborated method represents a rapid technique that can be used not only for the separation of proteins but also for analytical purposes.

Cromatografia de afinidade por íons metálicos imobilizados (IMAC) de biomoléculas: aspectos fundamentais e aplicações tecnológicas

“…[40][41][42][43][44][45][46] Outros materiais, como capilares monolíticos de sílica, nanotubos de carbono e fulerenos também são utilizados para a imobilização de íons metálicos na presença ou ausência de agentes quelantes. 47 Adsorventes IMAC (matriz com agente quelante imobilizado) podem ser também adquiridos comercialmente.…”

Lysozyme purification with dye-affinity beads under magnetic field

“…Magnetic separation promises to solve many of the problems associated with chromatographic techniques in packed bed and in conventional fluidized bed systems [17]. The magnetic character implies that they respond to a magnet, making sampling and collection easier and faster, but their magnetization disappears once the magnetic field is removed.…”

Use of magnetic poly(glycidyl methacrylate) monosize beads for the purification of lysozyme in batch system

“…They can be easily separated from reaction medium and stabilized in a fluidized bed column by applying a magnetic field. The use of magnetic beads reduces capital and operation costs [34]. The goal of this study is to prepare a tryptophan containing magnetic monosize poly(glycidyl methacrylate) [m-poly(GMA)] beads with magnetite nano-powder (i.e., Fe 3 O 4 ) for efficient separation of lysozyme from egg white.…”