Preparation and characterization of metal‐chelated poly(HEMA‐MAH) monolithic cryogels and their use for DNA adsorption

Abstract

DNA adsorption properties of Zn 2þ -chelated supermacroporous poly(2-hydroxyethyl methacrylate-Nmethacryloyl-(L)-histidine methyl ester) [poly(HEMA-MAH)] monolithic cryogel column were investigated for the application of DNA-affinity adsorbents. The monolithic cryogel was loaded with Zn 2þ ions to form the metal-chelated affinity sorbent. Poly(HEMA-MAH) cryogel was characterized by swelling tests, FTIR, scanning electron microscopy (SEM), and elemental analysis. SEM analysis indicates that the cryogel have a heteroporous structure with interconnected pores of 10-50 lm size, which ascribed to the porogens effect of frozen water crystals. Poly(HEMA-MAH) cryogel containing 45.8 lmol MAH was used in the adsorption/desorption of DNA from aqueous solutions. The maximum amount of DNA adsorption was 32.93 mg/g in Tris buffer at pH 7.0. It was observed that DNA could be repeatedly adsorbed and desorbed with the poly(HEMA-MAH) cryogel without significant loss of adsorption capacity. As a result, these higher amounts of DNA adsorbed poly (HEMA-MAH) cryogels are expected to be good candidate for achieving higher removal of anti-DNA antibodies from systemic lupus erythematosus (SLE) patients plasma.

Affinity monolith chromatography: A review of general principles and recent developments

“…Some of the monomers that have been used for cryogel preparation in AMC are agarose, HEMA, acrylamide, and 2‐(dimethylamino)ethyl methacrylate [14,44,136,138–146]. Affinity ligands that have been employed with cryogels include metal ions (e.g., Cu 2+ , Zn 2+ , and Ni 2+ bound via IDA), lectins (Con A), protein A, synthetic dyes (Cibacron Blue), and amino acids [136,138,139,141–153]. These binding agents have been applied to the separation and purification of yeast and bacteria [136,139,141,150], blood cells [153], general proteins (e.g., HSA, BSA, interferon, cytochrome c, and lactoferrin) [138,142,145,146,149,154–157], enzymes (e.g., chicken egg lysozyme, urokinase, and β‐glucosidase) [138,143,147,148,158], antibodies (IgM and IgG) [144,152,159], DNA [151], and glycoproteins (e.g., horseradish peroxidase) [160].…”

“…Affinity ligands that have been employed with cryogels include metal ions (e.g., Cu 2+ , Zn 2+ , and Ni 2+ bound via IDA), lectins (Con A), protein A, synthetic dyes (Cibacron Blue), and amino acids [136,138,139,141–153]. These binding agents have been applied to the separation and purification of yeast and bacteria [136,139,141,150], blood cells [153], general proteins (e.g., HSA, BSA, interferon, cytochrome c, and lactoferrin) [138,142,145,146,149,154–157], enzymes (e.g., chicken egg lysozyme, urokinase, and β‐glucosidase) [138,143,147,148,158], antibodies (IgM and IgG) [144,152,159], DNA [151], and glycoproteins (e.g., horseradish peroxidase) [160]. A cryogel monolith based on poly(vinyl alcohol)/polyethyleneimine and containing Cu 2+ has been used for the removal of hemoglobin from blood [161] and the carbohydrate N ‐acetyl‐D‐glucosamine was immobilized onto a cryogel monolith for the purification of Con A [162].…”

Molecular docking of metal ion immobilized ligands to proteins in affinity chromatography

“…32 MAH monomer was used as the metal-coordinating ligand due to the affinity of imidazole nitrogen donor atoms toward metal ions for the purification of various proteins and DNA in many years. 33,34 The 3D structure of MAH is not available in any database. Hence, this monomer was drawn and created using Avogadro software which has an auto-optimization tool ( Figure 3A).…”

Challenges and Advances in the Fabrication of Monolithic Bioseparation Materials and their Applications in Proteomics Research

“…Cryogels have interconnected macropores with pore sizes from 10 to 250 µm, demonstrating several merits such as low pressure drop, short diffusion path and retention time . They attracted great interests for fast enrichment and purification of biomacromolecules including enzymes, coenzymes, cofactors, antibodies, antigens, and DNA, as well as isolation of microbial cells and virus particles . For example, both plant‐derived human monoclonal antibody and glycosylated human monoclonal antibody were chemically immobilized onto poly(acrylamide‐ co ‐allyl glycidyl ether) cryogels by Sandeman and co‐workers The latter exhibited higher binding capacities of 117 mg g −1 toward anthrax toxin protective antigen than the former, indicating potential application of cryogel adsorbents for treatment of Bacillusanthracis infection.…”