Impact of double cryogelation process on a macroporous dye-affinity hydrogel

Cryogels with interconnected channels allow high flow-through properties and mass transfer when dealing with complex mixtures such as non-clarified crude extracts. However, their mechanical strength can be challenged due to a large void volume inside the polymeric network. We have addressed this problem by forming a double-layer cryogel applied as a dye-affinity chromatography gel. In this study, poly(acrylamide-co-allyl glycidyl ether) cryogel was prepared at sub-zero temperature. The second layer was then prepared inside the primary cryogel under the same conditions to form a double-layer network. Cibacron Blue F3GA, a dye molecule, was immobilized on the surface of the cryogels. Bovine serum albumin was used as a model molecule to study the adsorption/elution procedure in batch and continuous modes. The maximum batch binding capacity and the dynamic binding capacity for the single-layer cryogel were 18 and 0.11, and for the double-layer cryogel were 7.5 and 0.9 mg/g of gel, respectively. However, the mechanical stability of the double-layer cryogel increased 7-fold (144 kPa). It was found that the kinetic and adsorption isotherms follow pseudo-second-order and Freundlich models, respectively. The regeneration of the columns after adsorption/elution cycles was evaluated, and no significant loss of capacity was observed after 10 cycles.     

Hydrophobic cryogels for DNA adsorption: Effect of embedding of monosize microbeads into cryogel network on their adsorptive performances

As alternative hydrophobic adsorbent for DNA adsorption, supermacroporous cryogel disks were synthesized via free radical polymerization. In this study, we have prepared two kinds of cryogel disks: (i) poly(2‐hydroxyethyl methacrylate‐N‐methacryloyl‐l ‐tryptophan) [p(HEMA‐MATrp)] cryogel containing specific hydrophobic ligand MATrp; and (ii) monosize p(HEMA‐MATrp) particles synthesized via suspension polymerization embedded into p(HEMA) cryogel structure to obtain p(HEMA‐MATrp)/p(HEMA) composite cryogel disks. These cryogel disks containing hydrophobic functional group were characterized via swelling studies, Fourier transform infrared spectroscopy, elemental analysis, surface area measurements and scanning electron microscopy. DNA adsorption onto both p(HEMA‐MATrp) cryogel and p(HEMA‐MATrp)/p(HEMA) composite cryogels was investigated. Maximum adsorption of DNA on p(HEMA‐MATrp) cryogel was found to be 15 mg/g polymer. Otherwise, p(HEMA‐MATrp)/p(HEMA) composite cryogels significantly increased the DNA adsorption capacity to 38 mg/g polymer. Composite cryogels could be used repeatedly without significant loss on adsorption capacity after 10 repetitive adsorption–desorption cycles. Copyright © 2013 John Wiley & Sons, Ltd.     

Immobilization of Inulinase on Concanavalin A-Attached Super Macroporous Cryogel for Production of High-Fructose Syrup

In this study, concanavalin A (Con A)-attached poly(ethylene glycol dimethacrylate) [poly(EGDMA)] cryogels were used for immobilization of Aspergillus niger inulinase. For this purposes, the monolithic cryogel column was prepared by radical cryocopolymerization of EGDMA as a monomer and N,N′-methylene bisacrylamide as a crosslinker. Then, Con A was attached by covalent binding onto amino-activated poly(EGDMA) cryogel via glutaraldehyde activation. Characterization of cryogels was performed by FTIR, EDX, and SEM studies. Poly(EGDMA) cryogels were highly porous and pore size was found to be approximately 50–100 μm. Con A-attached poly(EGDMA) cryogels was used in the adsorption of inulinase from aqueous solutions. Adsorption of inulinase on the Con A-attached poly(EGDMA) cryogel was performed in continuous system and the effects of pH, inulinase concentration, and flow rate on adsorption were investigated. The maximum amount of inulinase adsorption was calculated to be 27.85 mg/g cryogel at 1.0 mg/mL inulinase concentration and in acetate buffer at pH 4.0. Immobilized inulinase was effectively used in continuous preparation of high-fructose syrup. Inulin was converted to fructose in a continuous system and released fructose concentration was found to be 0.23 mg/mL at the end of 5 min of hydrolysis. High-fructose content of the syrup was demonstrated by thin layer chromatography.     

Gelatin-Immobilised Poly(hydroxyethyl methacrylate) Cryogel for Affinity Purification of Fibronectin

In this work, fibronectin purification from human plasma with the gelatin-immobilised poly(hydroxyethyl methacrylate) (PHEMA) cryogel has been evaluated. The PHEMA cryogel was prepared by cryo-polymerisation which proceeds in an aqueous solution of monomer frozen inside a plastic syringe. The PHEMA cryogel contained interconnected macrochannels of 10–200 μm in diameter. Gelatin molecules were covalently immobilised onto the PHEMA cryogel via carbodiimide activation. The gelatin-immobilised PHEMA cryogel was used to purify fibronectin from human plasma. Fibronectin adsorption from human plasma on the PHEMA cryogel was 0.30 mg/ml, while much higher adsorption values, up to 38 mg/ml, was obtained with the gelatin-immobilised PHEMA cryogel. The fibronectin adsorption capacity of the gelatin-immobilised PHEMA cryogel did not change with an increase in the flow rate of plasma. Up to 92 % of the adsorbed fibronectin was eluted using 2 M urea containing 1 M NaCl as elution agent. The adsorption–elution cycle was repeated ten times using the same PHEMA cryogel. No remarkable decrease was detected in the adsorption capacity of the gelatin-immobilised PHEMA cryogel.     

In-situ graft-polymerization preparation of cation-exchange supermacroporous cryogel with sulfo groups in glass columns

Graft polymerization of monomer chains with expected functional groups onto the matrix pore surfaces by initiator is an effective approach for introducing ion-exchange groups to cryogel matrix to get anion- or cation-exchange supermacroporous cryogels. In this work, a novel cation-exchange cryogel with sulfo binding groups was prepared by grafting of 2-acrylamido-2-methyl-1-propanesulfonic acid (AMPSA) onto polyacrylamide-based cryogels in glass columns. The grafting polymerization was achieved in an in-situ manner which was performed by pumping the initiator and the reactive solution of graft monomer with sulfo binding groups directly through a cryogel bed pre-produced in a glass column under frozen condition. The axial liquid dispersion characteristics within the monolithic cryogel beds before and after the in-situ polymerization were compared by measuring residence time distributions (RTDs) at various liquid flow rates using tracer pulse-response method. Microstructure morphology of pores within cryogels was analyzed by scanning electron microscopy (SEM). Chromatography of lysozyme was carried out to reveal the protein breakthrough and elution characteristics in the obtained cryogel beds.     

DNA adsorption via Co(II) immobilized cryogels

The separation and purification of important biomolecule deoxyribonucleic acid (DNA) molecules are extremely important. The adsorption technique among these methods is highly preferred as the adsorbent cryogels are pretty much used due to large pores and the associated flow channels. In this study, the adsorption of DNA via Co(II) immobilized poly(2-hydroxyethyl methacrylate-glycidyl methacrylate) [poly(HEMA-GMA)] cryogels was performed under varying conditions of pH, interaction time, initial DNA concentration, temperature, and ionic strength. For the characterization of cryogels; swelling test, Fourier transform infrared (FT-IR) spectroscopy, scanning electron microscopy (SEM), surface area (BET), elemental and ICP-OES analysis were performed. L-lysine amino acid was chosen as Co(II)-chelating agent and the adsorption capacity of cryogels was determined as 33.81 mg DNA/g cryogel. Adsorption of pea DNA was studied under the optimum adsorption conditions and DNA adsorption capacity of cryogels was found as 10.14 mg DNA/g cryogel. The adsorption process was examined via Langmuir and Freundlich isotherm models and the Langmuir adsorption model was determined to be more appropriate for the DNA adsorption onto cryogels.     

Preparation of agarose/chitosan composite supermacroporous monolithic cryogels for affinity purification of glycoproteins

Supermacroporous agarose/chitosan composite monolithic (AC CM) cryogels were prepared for affinity purification of the major egg white glycoproteins, ovalbumin (OVA), and ovotransferrin (OVT). The supermacroporous AC CM cryogels were produced by cryocopolymerization of agarose/chitosan blend solutions using glutaraldehyde as the cross-linker. The 3-aminophenlyboronic acid ligand was immobilized by covalent binding to epoxy-group-coupled supermacroporous AC CM cryogels. The microstructure morphologies of these cryogels were analyzed by scanning electron microscopy. The supermacroporous AC CM cryogels contained a continuous interpenetrating polymer network matrix with interconnected pores of 10-100 μm in size. The composite cryogels offered high mechanical stability and had specific recognition for glycoproteins. The maximum binding capacity of OVA adsorption from aqueous solutions was 55.6 mg/g. The matrix could be reused 11 times without significant loss in OVA adsorption capacity. The recovery yields of OVA and OVT from egg white were estimated to be 89 and 93%, respectively.     

Fabrication of poly(lactide‐co‐glycolide) scaffold embedded spatially with hydroxyapatite particles on pore walls for bone tissue engineering

Poly(lactide‐co‐glycolide) (PLGA) scaffolds embedded spatially with hydroxyapatite (HA) particles on the pore walls (PLGA/HA‐S) were fabricated by using HA‐coated paraffin spheres as porogens, which were prepared by Pickering emulsion. For comparisons, PLGA scaffolds loaded with same amount of HA particles (2%) in the matrix (PLGA/HA‐M) and pure PLGA scaffolds were prepared by using pure paraffin spheres as porogens. Although the three types of scaffolds had same pore size (450–600 µm) and similar porosity (90%–93%), the PLGA/HA‐S showed the highest compression modulus. The embedment of the HA particles on the pore walls endow the PLGA/HA‐S scaffold with a stronger ability of protein adsorption and mineralization as well as a larger mechanical strength against compression. In vitro culture of rat bone marrow stem cells revealed that cell morphology and proliferation ability were similar on all the scaffolds. However, the alkaline phosphatase activity was significantly improved for the cells cultured on the PLGA/HA‐S scaffolds. Therefore, the method for fabricating scaffolds with spatially embedded nanoparticles provides a new way to obtain the bioactive scaffolds for tissue engineering. Copyright © 2011 John Wiley & Sons, Ltd.     

Lectin attached affinity cryogels for amyloglucosidase adsorption

Abstract

Preparation of poly[ethylene glycol dimethacrylate (EGDMA)-glycidyl methacrylate (GMA)] cryogel and its usability for amyloglucosidase adsorption were investigated. Cryogels were prepared by cryocopolymerization technique and then functionalized by lectin concanavalin A (Con A). The synthesized cryogel was characterized by FTIR, SEM and EDX analysis and then used for adsorption of amyloglucosidase in a continuous flow system. The maximal amyloglucosidase adsorption efficiency of Con A attached cryogel was found to be as 30.50?mg/g cryogel. Con A modified cryogels were used repeatedly for 30 times without any significant decrease at the amyloglucosidase adsorption capacity. SDS-PAGE and activity studies confirmed that the desorbed amyloglucosidase was active and retained 90% of its initial activity after the adsorption/desorption processes.     

Monolithic molecularly imprinted cryogel for lysozyme recognition

The application of molecularly imprinted polymers in the selective adsorption of macromolecules such as proteins by monolithic protein‐imprinted columns requires a macroporous structure, which can be provided by cryogelation at low temperature in which the formation of ice crystals gives a porous structure to the molecularly imprinted polymer. In this study, we applied this technique to synthesize lysozyme‐imprinted polyacrylamide cryogels containing 8% w/v of total monomers and 0.3% w/v of lysozyme. The synthesized cryogel was sponge‐like and elastic with very fast swelling and reshaping properties, showing a swelling ratio of 24.5 ± 3 and gel fraction yield of about 72%. It showed an imprinting effect of 1.58 and a separation factor of 1.37 for cytochrome c as the competing protein. Adsorption studies on the cryogel revealed that it follows the Langmuir isotherm, with a maximum theoretical adsorption capacity of 36.3 mg lysozyme per gram of cryogel. Additionally, it was shown that a salt‐free rebinding solution at low flow rate and pH = 7.0 is favorable for lysozyme rebinding. This kind of monolithic column promises a wide range of application in separation of various biomolecules due to its preparation simplicity, good rebinding characteristics, and macroporosity.     

Super macroporous poly(N‐isopropyl acrylamide) cryogel for separation purpose

Polymers that can respond reversibly by changing their physical or chemical properties are recognized as stimuli‐responsive polymers. The renowned temperature‐sensitive polymer is poly(N‐isopropyl acrylamide) (p(NIPAM)), and here, homopolymeric supermacroporous p(NIPAM)) cryogel was synthesized via cryopolymerization technique at cryogenic condition (below melting point of solvent, −18°C). Then, the prepared p(NIPAM) cryogel was characterized via scanning electron microscopy, Fourier transform infrared radiation spectrometer, and thermogravimetric analyzer. The lower critical solution temperature (LCST) value of the prepared p(NIPAM) cryogel was determined from % swelling equilibrium swellings at various temperatures, 20, 25, 30, 35, 40, 45, and 50°C, respectively. Furthermore, the pore volume and porosity of p(NIPAM) cryogels were compared below and above the LCST values. Finally, the separation capability of p(NIPAM) cryogels for some molecules such as tannic acid, gallic acid, nicotine (N), and caffeine (C) was investigated at the below and above the LCST values.     

Protein recognition via ion-coordinated molecularly imprinted supermacroporous cryogels

Molecular imprinting is a method for making selective binding sites in synthetic polymers using a molecular template. The aim of this study is to prepare lysozyme-imprinted supermacroporous cryogels which can be used for the purification of lysozyme (Lyz) from egg white. N-Methacryloyl-(L)-histidinemethylester (MAH) was chosen as the metal-coordinating monomer. In the first step, Cu2+ was complexed with MAH and the lysozyme-imprinted poly(HEMA-MAH) [Lyz-MIP] cryogel were produced by free radical polymerization initiated by N,N,N',N'-tetramethylene diamine (TEMED) in an ice bath. After that, the template (i.e., lysozyme) was removed using 0.05 M phosphate buffer containing 1M NaCl (pH 8.0). The maximum lysozyme adsorption capacity was 22.9 mg/g polymer. The relative selectivity coefficients of Lyz-MIP cryogel for lysozyme/bovine serum albumin and lysozyme/cytochrome c were 4.6 and 3.2 times greater than non-imprinted poly(HEMA-MAH) (NIP) cryogel, respectively. Purification of lysozyme from egg white was also monitored by determining the lysozyme activity using Micrococcus lysodeikticus as substrate. The purity of the desorbed lysozyme was about 94% with recovery about 86%. The Lyz-MIP cryogel could be used many times without decreasing the adsorption capacity significantly.     

A Study of cryostructuring of polymer systems. 41. Complex and composite poly(vinyl alcohol) cryogels containing soluble and insoluble forms of chitosan,respectively

Complex macroporous poly(vinyl alcohol) (PVA) cryogels have been obtained by cryogenic treatment (freezing at–20°C for 12 h followed by defrosting at a rate of 0.03°C/min) of PVA–chitosan hydrochloride mixed solutions. The subsequent alkaline treatment of the cryogels has resulted in the transformation of the water-soluble salt form of chitosan into its insoluble basic form, which coagulates inside the bulk of the continuous phase of PVA cryogel into small particles with sizes of 2–5 µm. In the resulting composite cryogels, these particles play the role of an “active” filler, which increases the rigidity and heat endurance of the gel material. It has been shown that the sorption capacity of such chitosan particles entrapped into the bulk of composite cryogels with respect of bivalent copper ions is noticeably higher than the sorption capacity of ground chitosan particles incorporated as a discrete filler into the continuous phase PVA cryogels. The study of the properties of PVA–chitosan hydrochloride mixed solutions revealed that these polymers are, to a large extent, compatible with one another in a common solvent at a low ionic strength. Therefore, liquidliquid phase separation of these systems due to the thermodynamic incompatibility of macromolecules of different natures is observed only upon increasing the ionic strength by adding a low-molecular-mass salt (NaCl, 0.15 mol/L) to the solution.     

A Crosslinked Carboxylic Acid Containing Cation Exchange Monolithic Cryogel for Human Serum Albumin Separation

For this work, we synthesized poly(N-isopropylacrylamide-acrylamide)-acrylic acid (poly(NIPAM-Am)-AAc) monolithic cryogel for a human serum albumin separation (HSA) from a protein mixture (human serum immunoglobulin, human serum albumin and lysozyme) and performed HSA adsorption studies using the cryogel to do continuous system experiments in a syringe column connected by a peristaltic pump. Poly(NIPAM-Am)-AAc with a pore size of 10–100 μm was produced by free radical polymerization that proceeded in an aqueous solution of monomers frozen inside a syringe column. The monolithic poly(NIPAM-Am)-AAc cryogel was characterized by performing swelling studies, FTIR and SEM that showed a swelling ratio of 6.2 g H₂O/g dry cryogel. The maximum HSA adsorption by the cryogel was 42.5 mg/g polymer at pH 4.0 in a 50 mM acetate buffer. We also studied the effect of two different temperatures (25 and 40°C). The higher temperature increased the adsorption capacity of the cryogel. HSA molecules could be reversibly adsorbed and desorbed five times with the same poly(NIPAM-Am)-AAc cryogel without a noticeable loss of their HSA adsorption capacity. The synthesized cryogel was used to separate albumin from the protein mixture. Adsorbed albumin was eluted by changing the pH of the buffer (pH 7.0 and 25°C). Poly(NIPAM-Am)-AAc monolithic cryogel behaved as a cation exchange column because of its functional carboxylic group.     

PolyGuanine methacrylate cryogels for ribonucleic acid purification

The isolation and purification of ribonucleic acid have attracted attention recently for the understanding of the functions in detail because of the necessity for the treatment of genetic diseases. In this study, guanine‐incorporated polymeric cryogels were developed to obtain highly purified ribonucleic acid. The satisfactory purification performance was achieved with the guanine‐incorporated poly (2‐hydroxyethyl methacrylate‐guanine methacrylate) cryogels. The most crucial advantages to use guanine as a functional monomer are to obtain a real natural interaction between guanine on the polymeric material and cytosine on the ribonucleic acid. Moreover, using cryogel with a highly porous structure and high swelling ratio provide advantages of getting more water within the structure to get more analyte to interact. The characterization of cryogels has proved the success of the synthesis and the perfect natural interaction to be taken place between the ligand (guanine methacrylate) and the cytosine in the ribonucleic acid molecules. Although the pores within the structure of cryogels are small, they provide efficient and fast adsorption. The chromatographic separation performance was investigated for different conditions (pH, temperature etc.). The desorption ratio and reusability were also analyzed at the end of the five adsorption–desorption cycles with no significant changes.     

Specific features of the polyelectrolyte behavior of weakly charged cryogels of polyacrylamide and poly(N-isopropylacrylamide)

Specific features of the polyelectrolyte behavior of weakly charged common gels and cryogels of copolymers of polyacrylamide and poly(N-isopropylacrylamide) with sodium acrylamido-2-methyl-1-propyl sulfonate are investigated. The cryogels are synthesized in frozen solutions at ?15°C. It is shown that the polyelectrolyte swelling is significantly weaker in the case of cryogels than that in the case of gels synthesized in solutions. For thermosensitive gels with isopropylacrylamide groups, collapse occurs during heating. Charging of a common gel leads to a noticeable (18°C) increase in the transition temperature. For a cryogel, this growth is 3°C. During the interaction with cetylpyridinium chloride, the gel contraction is much more pronounced for common weakly charged gels. At the same time, walls of pores of a collapsed cryogel contain a smaller amount of the solvent. Isotherms of the adsorption of a cationic surfactant by anionic common gels and cryogels differ insignificantly. Model gels synthesized in concentrated acrylamide solutions exhibit very weak polyelectrolyte swelling, similar to that of cryogels. The behavior of cryogels is explained by a very high local concentration of crosslinks due to a strong entanglement of polymer chains.     

Poly(HEMA-co-NBMI) Monolithic Cryogel Columns for IgG Adsorption

Supermacroporous poly(2-hydroxyethyl methacrylate-co-1,5-naphthalene bismaleimide) [poly(HEMA-co-NBMI)] monolithic cryogel column was prepared by free radical cryo-copolymerization of HEMA with NBMI as a hydrophobic functional comonomer and N,N′-methylene-bisacrylamide as cross-linker directly in a plastic syringe for adsorption of albumin. The monolithic cryogel contained a continuous polymeric matrix which has interconnected pores of 10–100 μm size. Poly(HEMA-co-NBMI) cryogel was characterized by swelling studies, FTIR and scanning electron microscopy. The equilibrium swelling degree of the poly(HEMA-co-NBMI) cryogel was 10.5 g of H₂O/g dry cryogel. Poly(HEMA-co-NBMI) cryogel was used in the adsorption/desorption of IgG from aqueous solutions. The maximum amount of IgG adsorption from aqueous solution in phosphate buffer was 98.20 mg/g polymer at pH 7.0. The nonspecific adsorption of IgG onto plain poly(HEMA) cryogel was very low (2.79 g/g polymer). It was observed that IgG could be repeatedly adsorbed and desorbed with the poly(HEMA-co-NBMI) cryogel without significant loss of adsorption capacity.     

Oriented immobilized anti‐hIgG via Fc fragment‐imprinted PHEMA cryogel for IgG purification

Antibodies are used in many applications, especially as diagnostic and therapeutic agents. Among the various techniques used for the purification of antibodies, immunoaffinity chromatography is by far the most common. For this purpose, oriented immobilization of antibodies is an important step for the efficiency of purification step. In this study, F_c fragment‐imprinted poly(hydroxyethyl methacrylate) cryogel (MIP) was prepared for the oriented immobilization of anti‐hIgG for IgG purification from human plasma. Non‐imprinted poly(hydroxyethyl methacrylate) cryogel (NIP) was also prepared for random immobilization of anti‐hIgG to compare the adsorption capacities of oriented (MIP/anti‐hIgG) and random (NIP/anti‐hIgG) cryogel columns. The amount of immobilized anti‐hIgG was 19.8 mg/g for the NIP column and 23.7 mg/g for the MIP column. Although the amount of immobilized anti‐hIgG was almost the same for the NIP and MIP columns, IgG adsorption capacity was found to be three times higher than the NIP/anti‐hIgG column (29.7 mg/g) for the MIP/anti‐hIgG column (86.9 mg/g). Higher IgG adsorption capacity was observed from human plasma (up to 106.4 mg/g) with the MIP/anti‐hIgG cryogel column. Adsorbed IgG was eluted using 1.0 m NaCl with a purity of 96.7%. The results obtained here are very encouraging and showed the usability of MIP/anti‐hIgG cryogel prepared via imprinting of Fc fragments as an alternative to conventional immunoaffinity techniques for IgG purification. Copyright © 2012 John Wiley & Sons, Ltd.     

Graft polymerization of vinyl monomers inside macroporous polyacrylamide gel,cryogel, in aqueous and aqueous‐organic media initiated by diperiodatocuprate(III) complexes

Graft polymerization initiated by diperiodatocuprate(III) complex (Cu(III)) initiator was found to be an effective and convenient method for graft polymerization of vinyl monomers onto macroporous polyacrylamide gels, the so‐called cryogels (pAAm‐cryogels). The effect of time, temperature, monomer and initiator concentration during the graft polymerization in aqueous and aqueous‐organic media was studied. The graft polymerization of water‐soluble monomers as [2‐(methacryloyloxy)ethyl]‐trimethylammonium chloride, 2‐hydroxyethyl methacrylate, N‐isopropylacrylamide, and N,N‐dimethylacrylamide proceeds with higher grafting yield in aqueous medium, as compared with that in aqueous‐organic media. Graft polymerization in aqueous‐organic media such as water–DMSO solutions allows grafting of water‐insoluble monomers such as glycidyl methacrylate and N‐tert‐butylacrylamide with high grafting degrees of 100 and 410%, respectively. It was found that the deposition of initiator on the pore surface of cryogels promoted graft polymerization by facilitating the formation of the redox couple Cu(III)‐acrylamide group. © 2006 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 44: 1952–1963, 2006     

Histidine-epoxy-activated sepharose beads embedded poly (2-hydroxyethyl methacrylate) cryogels for pseudobiospecific adsorption of human immunoglobulin G

The use of highly purified immunoglobulin became among the most powerful adopted strategies in therapeutic trials nowadays. Their role as immunomodulatory and anti-inflammatory agents has widened their scope of use. A novel continuous supermacroporous monolithic cryogels embedded with histidine-epoxy-activated-sepharose beads were synthetized as a new monolithic adsorbents for the separation of immunoglobulin G from human serum. The histidine-epoxy-activated-sepharose beads were embedded into the 2-hydroxyethyl methacrylate (HEMA) cryogels present in frozen aqueous solution inside a plastic syringe. The microstructure morphology of the cryogels was characterized by swelling measurement and scanning electron microscopy. The adsorption of human IgG on the histidine-epoxy-activated-sepharose beads pHEMA cryogels appeared to follow the Langmuir–Freundlich adsorption isotherm model. The maximum IgG adsorption was observed at 4°C and pH 7.4 and was found to be 26.95 mg/g of cryogel which is close to that obtained experimentally (24.49 mg/g). The cryogels were used for several adsorption-desorption cycles without any negligible decrease in their adsorption capacity.