Synthesis of nonlinear polymer brushes on magnetic nanoparticles as an affinity adsorbent for His-tagged xylanase purifi
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Synthesis of nonlinear polymer brushes on magnetic nanoparticles as an affinity adsorbent for His-tagged xylanase purification Zahra Shirzadi 1 & Habibollah Baharvand 2 & Mahshid Nikpour Nezhati 1 & Reza H. Sajedi 3 Received: 10 June 2020 / Revised: 25 August 2020 / Accepted: 3 September 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract In this article, the magnetic nanospheres bonded to a polymer chelator have synthesized through a new strategy consisted of four parts: (1) synthesis and surface modification of nanoparticles with vinyl groups; (2) grafting of a water-soluble polymer having hydroxyl groups, by polymerization of the resulting monomer from the reaction of glycidyl methacrylate (GMA) and diethanolamine (DEA) on the surface of nanomagnetic particles; (3) by cerium(IV), conversion of hydroxyl groups of the synthesized polymer on the nanoparticles to radicals for subsequent grafting of the resulting monomer from reaction of GMA and iminodiacetic acid (IDA); and (4) in an ion-exchange process, nickel ions along the last synthesized polymer were immobilized. The prepared nanospheres were successfully used directly for purification of His-tagged xylanase from cell lysates. The separated proteins were subjected to enzyme assays and are observed well in the enzymatic activity. Keywords Fe3O4 nanospheres . Purification . His-tagged proteins . Enzyme immobilization . Xylanase
Introduction Recombinant proteins are now widely used in the proteomics [1] and pharmaceutical industries [2–4], so the production and purification of these proteins are of particular importance. One of the successful methods for purification of His-tagged recombinant proteins is the use of immobilized metal affinity chromatography (IMAC) [5]. The basis of this approach is relied on the interactions between the immobilized metal ions and the electron donor groups like the histidine groups on the protein surface. There are some problems with using this technique, such as long process time, low process efficiency, protein solubility, and the need to isolate colloidal contaminants and cell debris from the environment. This technique is ineffective especially when the target protein concentration in a
* Habibollah Baharvand [email protected] 1
Department of Chemistry, Islamic Azad University, Central Tehran Branch, Tehran, Iran
2
Faculty of Polymer Science, Iran Polymer and Petrochemical Institute, P.O. Box: 112/14975, Tehran, Iran
3
Department of Biological Science, Tarbiat Modares University, Tehran, Iran
limited sample is very low and needs to be enriched at high concentrations [6, 7]. Given the possibility of separation of magnetic particles from a solution by an external magnetic field, nowadays, with proper design of the surface of these particles, their chemical modification with organic or inorganic materials is widely used in bio-separations without any previously sample preparation [8–15]. In addition, this technique can be used when the sample size is very limited, and it
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