Synthesis and performance of 3D‐Megaporous structures for enzyme immobilization and protein capture

Abstract

The preparation of megaporous bodies, with potential applications in biotechnology, was attempted by following several strategies. As a first step, naive and robust scaffolds were produced by polymerization of selected monomers in the presence of a highly soluble cross-linker agent. Ion-exchange function was incorporated by particle embedding, direct chemical synthesis, or radiation-induced grafting. The total ionic capacity of such systems was 1.5 mmol H(+)/g, 1.4 mmol H(+)/g, and 17 mmol H(+)/g, respectively. These values were in agreement with the ability to bind model proteins: observed dynamic binding capacity at 50% breakthrough was ≅7.2 mg bovine serum albumin/g, ≅7.4 hen egg-white lysozyme (HEWL) mg/g, and ≅108 HEWL mg/g. In the later case, total (static) binding capacity reached 220 mg/g. It was observed that the structure and size of the megapores remained unaffected by the grafting procedure which, however, allowed for the highest protein binding capacity. Lysozyme supported on grafted body showed extensive clarification activity against a Micrococcus lysodekticus suspension in the flow-through mode, i.e., 90% destruction of suspended microbial cells was obtained with a residence time ≈ 18 min. Both protein capture and biocatalysis applications are conceivable with the 3D-megaporous materials described in this work.

Evaluation of the effectiveness of microparticle-embedded cryogel system in removal of 17β-estradiol from aqueous solution

“…Due to the high porosity, high permeability, soft, flexibility, and high flow velocity cryogels are widely used as new chromatographic materials for separation and purification of biomolecules, such as proteins, plasmid DNA, and viruses, from crude feedstocks [15][16][17] and also different applications as immobilization matrices in biotechnology [18][19][20][21], as scaffolds in tissue engineering [22][23][24][25][26], and as drug-delivery carriers in pharmaceutical fields [27]. Cryogels permit the free passage of microparticles, nanoparticles, or bioparticles without blockage because of the pore size within the range of 10-100 μm.…”

Current strategies and challenges for the purification of stem cells

“…56 Moreover, there is a current trend towards the substitution of antibody markers by novel aptamers-short oligonucleotides that serve as capture molecules. 15 The use of aptamers seems to be more advantageous due to the molecular flexibility that enables the molecule to bind with target sites, which would not be accessible for typical antibodies.…”

Preparation, characterization, and process performance of composite fibrous adsorbents as cation exchangers for high throughput and high capacity bioseparations

“…After extensive washing with water, the functionalized sulfonate group composite fibrous adsorbent (SP gPore) was dried and then the degree of grafting (DG) was determined by gravimetrically [5]. The material porosity by water uptake (porosity, expressed as %) and the degree of swelling (DS) of the SP gPore material were determined by the mentioned procedure [30].…”

“…To determine the existence of combined mass transfer resistance and dispersion effects and their potential influence on performance, dynamic binding capacity was evaluated via breakthrough curve (BTC) analysis as a function of superficial velocity. Moreover, residence time distribution (RTD) experiments utilizing acetone as an inert tracer were performed also as a function of flow velocity [30]; normalized profiles were interpreted by the dispersion model. The results of the breakthrough experiments carried out in a 1 mL column (5 mm ID × 5.5 cm L) as expressed by the quantities DBC and Peclet number (Pe) are shown in Fig.…”