Preparation of a novel metal‐chelate affinity beads for albumin isolation from human plasma

Abstract

ABSTRACT:In this study, we developed a novel approach to obtain a high protein-adsorption capacity utilizing 2-methacryloylamidohistidine (MAH) as a biollgand. MAH was synthesized by reacting methacryloyl chloride and histidine. Spherical beads, with an average size of 150 -200 m, were obtained by the radical suspension polymerization of MAH, ethyleneglycol dimethacrylate (EGDMA), and 2-hydroxyethyl methacrylate (HEMA) conducted in an aqueous dispersion medium. p(EGDMA-HEMA-MAH) beads had a specific surface area of 17.6 m 2 /g. The synthesized MAH monomer was characterized by NMR. p(EGDMA-HEMA-MAH) beads were characterized by a swelling test, FTIR, and elemental analysis. Then, Cu(II) ions were incorporated into the beads and Cu(II) loading was found to be 0.96 mmol/g. These beads, with a swelling ratio of 65% and containing 1.6 mmol MAH/g, were used in the adsorption/ desorption of human serum albumin (HSA) from both aqueous solutions and human serum. The adsorption of HSA onto p(EGDMA-HEMA-MAH) was low (8.8 mg/g). Cu(II) chelation onto the beads significantly increased the HSA adsorption (56.3 mg/g). The maximum HSA adsorption was observed at pH 8.0 Higher HSA adsorption was observed from human serum (94.6 mg HSA/g). Adsorptions of other serum proteins were obtained as 3.7 mg/g for fibrinogen and 8.5 mg/g for ␥-globulin. The total protein adsorption was determined as 107.1 mg/g. Desorption of HSA was obtained using a 0.1M Tris/HCI buffer containing 0.5M NaSCN. High desorption ratios (to 98% of the adsorbed HSA) were observed. It was possible to reuse Cu(II)-chelated p(EGDMA-HEMA-MAH) beads without significant decreases in the adsorption capacities.

Preparation of Zn2+‐chelated poly(HEMA‐MAH) cryogel for affinity purification of chicken egg lysozyme

“…In respect of ease preparation, economy, adsorption capacity, and stability of immobilized metal affinity chromatography (IMAC) offer several advantages over other conventional adsorbents 5–9. IMAC is a sensitive technique for protein separation that enables distinguishing between proteins differing by only a single histidine residue on the surface 10–14. It is assumed that proteins interact mainly through the imidazole group of histidine and, to a lesser extent, the indole‐group of tryptophan and the thiol group of cysteine.…”

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

“…A similar approach has been described for the study of subtilisin Carlsberg and BSA binding to Cu(II)-loaded magnetic microspheres [17,23,24]. IDA, the most popular metal-chelating ligand in IMAC, has been used as a metal-chelating ligand for binding to magnetic microspheres.…”

The usage of composite nanomaterials in biomedical engineering applications

“…These methacrylated based nanosized materials can be shaped in different forms such as cryogels (Avcibaşi et al, 2010; Perçin et al, 2013; Savina et al, 2007), membranes (Arica & Hasirci, 1993; Demir et al, 2018; Kermis et al, 2003) or beads (Akgöl et al, 2007; Jayakrishnan & Thanoo, 1990; Vianna‐Soares et al, 2004). They can be integrated to nanosized materials to seperate and isolate biomolecules with using different affinity chromatography techniques such as metal chelate (Karakoc et al, 2009; Odabasi et al, 2003), dye (Mislovičová et al, 1995; Unsal et al, 2005; Yavuz et al, 2006), boronate (Kuru et al, 2014; Srivastava et al, 2012; Toprak et al, 2015), lectin (Bereli et al, 2005; M.‐X. Hu & Xu, 2011; Kusat et al, 2020; Ulucan‐Karnak & Akgol, 2021) and so on.…”