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Serial numbers have forensic value as they help to identify firearms. While the serial numbers are mostly stamped on the metal parts of firearms, the characters on polymer parts involve non-serial number information such as caliber, brand, model, or proof marks, which also serve for identification purposes. The forensic evidence indicates that the polymer frames of firearms bearing non-serial number information are obliterated through heating and scraping. Although the destructive restoration techniques for polymers are well-documented, there is little theoretical and practical knowledge regarding the non-destructive restoration techniques applied on polymers. In view of this gap, this study aims to devise a non-destructive spectral technique to recover the obliterated characters on Polyamide 6. Considering its wide use on polymers, the numbering is carried out by hot stamping and the numbers are defaced through heating, scraping, and hammering both superficially and deeply at varying depths. Herein, we focused on imitating the manual obliteration techniques used by criminals instead of the deepness-controlled techniques preferred by previous studies. The samples are then viewed under the UV and IR lights in the Video Spectral Comparator 8000 for the first time. The results suggest that spectral imaging provided restoration to a good extent after heating and scraping which made the characters invisible at a relatively low depth of deformation compared to hammering. The recovery of characters with this novel technique brings a new perspective forensic marks examination literature by producing quick, successful and reliable results and facilitating reexamination by not harming the sample.

Ochratoxin A (OTA), a group of highly toxic fungal secondary metabolites, is one of the most common mycotoxins that contaminate a wide range of agricultural products. Herein, we designed a label-free and selective surface plasmon resonance (SPR) based sensor for detection of OTA contamination in dried fig. The molecularly imprinted polymer film (MIP) was fabricated onto the SPR sensor chip by using light-initiated polymerization of N-methacryloyl-(L)-phenylalanine and 2-Hydroxyethyl methacrylate in the presence of OTA as a template. SPR chips were characterized by using contact angle, atomic force microscopy, and ellipsometry. OTA imprinted sensor was displayed a broad linear range, between 0.1 ng/mL and 20 ng/mL. The limit of detection was calculated as 0.028 ng/mL. The selectivity studies of the OTA imprinted SPR sensor was performed in the presence of aflatoxin M1, aflatoxin B1, and citrinin. The imprinting factor was found to be 2.85. The adsorption behavior of the OTA imprinted sensor was found as suitable for the Langmuir model. Results displayed that the OTA imprinted SPR sensor was successfully applied to determine OTA in dried fig samples.

In this study, poly(2-hydroxyethyl methacrylate) [p(HEMA)] based hydrogels responsive to the pH, temperature and magnetic field were synthesized. The surface properties of p(HEMA) were improved by designing the stimuli-responsive hydrogels made of MAGA, NIPAAm and methacrylate-decorated magnetite nanoparticles as a function of pH-, thermo- and magnetic responsive cell culture surfaces. These materials were then modified an abundant extracellular matrix component, type I collagen, which has been considered as a biorecognition element to increase the applicability of hydrogels to cell viability. Based on results from scanning electron microscopy (SEM) and thermal gravimetric analysis (TGA), stimuli-responsive hydrogel demonstrated improved non-porous structures and thermal stability with a high degree of cross-linking. Mechanical analyses of the hydrogels also showed that stimuli-responsive hydrogels are more elastomeric due to the polymeric chains and heterogeneous amorphous segments compared to plain hydrogels. Furthermore, surface modification of hydrogels with collagen provided better biocompatibility, which was confirmed with L929 fibroblast cell adhesion. Produced stimuli-responsive hydrogels modulated cellular viability by changing pH and magnetic field.

In this study, a nanoparticle-based sandwich-like immunoassay was designed in dispersion medium to precisely detect apoptosis over caspase antibodies in order to overcome the disadvantages of traditional apoptosis determination methods such as high cost, large sampling requirement, and appropriate laboratory and equipment conditions. For this purpose, a complementary particulate system including magnetic (MNPs) and upconversion silica (UC-SiNPs) nanoparticles while immobilizing antibodies (primary antibody to MNPs, secondary antibody to UC-SiNPs) were synthesized and characterized. Optimization and selectivity studies of the complex formed by primary antibody immobilized MNPs with standard caspase proteins were examined by the HPLC system. Within the scope of optimization studies, protein concentrations, optimal duration, and temperature parameters were evaluated. Optimal conditions were determined for pH, initial concentration, time, and temperature as 7.4, 5.6 μg/mL, 45 min, and room temperature, respectively. Furthermore, the adsorption of competitive proteins was investigated in selectivity studies as well. Moreover, the primary antibody immobilized MNPs were treated with standard caspase proteins under optimal conditions; subsequently, they were interacted with secondary antibody immobilized UC-SiNPs to demonstrate the supracomplex formation meanwhile zeta potential/size measurements and fluorescence emission spectrometry analyses were performed. As a result of these analyses, it was observed that the sandwich-like supracomplexes were successfully formed that significantly varied upconversion emission intensities of UC-SiNPs in dependence on the amounts of caspase proteins. Because this approach enabled a quantitative result, the nanoparticle-based sandwich-like immunoassay should be classified as an easy-to-handled, fast, and promising alternative to benchmark apoptosis assays.

In this study, a new approach for the detection of benzo[a]pyrene (BaP) in aqueous media is presented by combining molecularly imprinted polymers (MIPs) with surface plasmon resonance (SPR) sensors. Herein, BaP imprinted sensor was fabricated on a SPR chip in the presence of a pre-polymerization complex of BaP and N-methacryloyl-L-phenylalanine, 2-hydroxyethyl methacrylate (HEMA), and ethylene glycol dimethacrylate (EGDMA). The developed sensor was characterized by atomic force microscopy, ellipsometry, and contact angle measurements. Association kinetics analysis via Scatchard, Langmuir, Freundlich, Langmuir–Freundlich isotherms were applied to the data and Langmuir adsorption model was found as the most accurate model for the affinity sensor system. MIPs-based SPR sensor linearly responded to BaP in a concentration range of 20-400 ng mL−1 with an excellent LOD of 14.97 ng L−1. The SPR sensor was found to be highly selective toward BaP according to the selectivity study results in the presence of interfering agents such as benzo[b]floranthene (BbF), benzo[k]floranthene (BkF), indeno[1,2,3-cd]pyrene (IcdP), and 1-naphthol. The studies demonstrated that MIPs-based SPR sensor provides rapid and reliable detection and identification of polyaromatic hydrocarbons (PAHs) within a quite short time.

In this study, we introduced a new strategy to design interface imprinted polymers for a novel aspect of molecular imprinting technique, utilization of sacrificial metal oxide particles. In the first step, bovine hemoglobin (BHb) was adsorbed on zinc oxide (ZnO) particles, which were then used to synthesize polyacrylic acid-based molecular imprinting membrane by bulk polymerization in the presence of ethylene glycol dimethacrylate as a cross-linking agent. After polymerization terminated, BHb-ZnO particles were removed to leave effective imprint sites onto the bulk polymeric network which is responsible for the formation of template orientation. The characterization of membranes was investigated by using Fourier transform infrared (FTIR), Raman spectroscopy (RS), scanning electron microscopy (SEM), surface area measurements (BET analyses) and thermogravimetric analysis (TGA). The interface molecularly imprinted membranes (iMIMs) have a relatively high specific rebinding capacity of 65.98 mg/g and excellent selectivity towards BHb with a separation factor of 6.78. The equilibrium adsorption isotherms fitted well to Langmuir isotherms (R2 = 0.9944) and the value of adsorption capability (Qmax) and equilibrium constant (b) were estimated to be 73.53 mg/g and 1.36 mg/mL for the iMIM, respectively. The kinetics of adsorption fitted best to pseudo-second order (R2 = 0.9912). The ZnO particles were used not only to ensure the preservation of the imprint cavities in the polymer network but also to lead to high template removal and better rebinding kinetics. This novel design with multiple recognition sites is quite simple and suitable for the separation of biomacromolecules.

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 tha...

In this study, cryogel-based molecularly imprinted composite cartridges were designed for the rapid, efficient, and selective preconcentration of benzo[ a ]pyrene (BaP) from water samples. First, a BaP-imprinted poly(2-hydroxyethyl methacrylate-N-methacryloyl-(L)-phenylalanine) composite cartridge was synthesized under semi-frozen conditions and characterized by scanning electron microscopy, elemental analysis, Fourier transform infrared spectroscopy, and swelling tests. After the optimization of preconcentration parameters, i.e., pH and initial BaP concentration, the selectivity and preconcentration efficiency, and reusability of these cartridges were also evaluated. In selectivity experiments, BaP imprinted composite cartridge exhibited binding capacities 3.09, 9.52, 8.87, and 8.77-fold higher than that of the non-imprinted composite cartridge in the presence of competitors, such as benzo[ b ]fluoranthene (BbF), benzo[ k ]fluoranthene (BkF), indeno[1,2,3- cd ]pyrene (IcdP), and 1-naphthol, respectively. The method detection limit (MDL), relative standard deviation (RSD) and preconcentration efficiency (PE) of the synthesized composite cartridge were calculated as 24.86 μg/L, 1.60%, and 349.6%, respectively.

Two-dimensional HPLC (2D-HPLC) recently has received great attention due to providing high resolving power and higher peak capacities than that of 1D-HPLC, especially dealing with a wide spectrum of sample matrices containing several components. In this work, an on-line heart-cutting two-dimensional liquid chromatography (2D-LC) method was developed using monolithic columns coupled with reversed-phase liquid chromatography (RPLC). 2D-HPLC was successfully carried out using affinity-based monolithic columns at first dimension (20 cm x 4.6 mm I.D.) followed by a Pinnacle II PAH column (50 mm x 4.6 mm I.D.) at the second dimension. Furthermore, good linearity was observed for the correlation of the benzo[a]pyrene (BaP) molecule against the peak areas (R-2 = 0.994) in the concentration range of 0.01-1.0 ng/mL in 30 min. The limit of detection and the limit of quantification were found to be 4.0 pg/mL and 12.0 pg/mL, respectively. Both the intra-day and inter-day precision at 0.01 and 0.1 ng/mL spiked concentrations were below than 2.35% RSD whereas the means of the recovery data of the BaP from the water samples were found to be in the range of 93.71-98.65%. These results demonstrate that the 2D-HPLC system, developed by the combination of the P(HEMA-MAPA) column and Pinnacle II PAH column, is reliable, stable, and well qualified in the extraction of polycyclic aromatic hydrocarbons from the water samples.

The continuous bed bioreactor systems have been used for the production of protein therapeutics, such as IgG, using immobilized enzyme in biopharmaceutical applications. We developed macroporous poly(hydroxyethyl methacrylate-co-glycidyl methacrylate) cryogel-based bioreactor matrix using sodium dodecyl sulfate as surfactans in the presence of ethylene glycol dimethacrylate as cross linking agent by bulk polymerization. The developed polyGMA immobilized bioreactor with papain enzyme was used for specific fragmentation of immunoglobulin G. The catalysis efficiency for immobilized enzyme were investigated in comparison with free enzyme. The immobilized papain displayed broad catalytic activity over a variety of conditions, with maximal activity around pH 7.0 and 70 °C. The Michaelis-Menten kinetic constant (Km), the maximum reaction velocity (Vmax), and the catalytic efficiency (kcat) for free enzyme were 0.1097 mg/mL, 29.9 mg/mL/min, and 92.01 1/min, respectively, whereas for immobilized enzyme, Km, Vmax, and kcat values were 0.1078 mg/mL, 30.53 mg/mL/min, and 94.3 1/min, respectively. In a further step, after digestion, remarkable digestion products of bioreactor, Fab and Fc fragments, produced with immobilized papain bioreactors were analyzed in two ways by SDS-PAGE and reversed-phase HPLC; it was demonstrated that papain immobilized bioreactor successfully used for the digestion of human IgG with high activity. Therefore, the polyGMA cryogel immobilized with papain exhibited a very effective matrix for the bioreactor which can be considered as an alternative bioreactor matrix with great promise in biopharmaceutical applications.

Immunoaffinity chromatography (IAC) is a kind of bioaffinity chromatography which used antibodies or antibody-related molecules as the stationary phase. IAC is used by many applications for analytical, clinical and diagnostic purposes, particularly preferring in analytical purposes on one-step separation and purification of target compounds. Moreover, immunoaffinity chromatography is used in antibody enrichment and separation of cells. IAC columns are usually applied in the antibody experiments due to powerful and selective binding of antibodies and/or their target antigens. Antigen or antibody molecules could be immobilized to the solid support. Therefore, target antibody or cell is purified. Specific bioligands can be immobilized directly on glycidyl based polymeric material with simple acid–base catalyst. In this study, polyglycidyl methacrylate based therefore cryogels were prepared and anti-insulin antibodies were immobilized on porous surface of cryogels. Swelling test, Fourier transform infrared spectroscopy (FTIR), Raman spectroscopy, and scanning electron microscopy (SEM) were conducted to characterize cryogels developed. To optimize separation conditions, effects of pH, initial insulin concentration, flow rate, salt concentration, contact time and temperature on insulin adsorption capacity were examined. The results indicated that the immunoaffinity cryogel developed here could be classified as good alternative with prominent properties such as high reusability and cost-friendly adsorbent and would be one of the primary reports for immunoaffinity purification of insulin molecules in not only lab-scale but also for industrial purposes.

WOS: 000354965300034, In this study, we synthesised specific filtration cartridges with selective recognition sites for target molecules and used them to separate interferon alpha-2b from aqueous solutions. We combined molecular imprinting technology with cryogel to achieve specific and rapid filtration of interferon alpha-2b through the macroporous structure of a cryogel network. Recombinant interferon alpha-2b-imprinted poly(2-hydroxyethyl methacrylate-N-methacryloyl-L-tryptophan) P(HEMATrp)/alpha-2bIFN cryogels were synthesised via free-radical bulk polymerisation under partially frozen conditions. The interferon alpha-2b filtration conditions were subsequently optimised with respect to factors such as pH, initial concentration, temperature, centrifugation speed, salt concentration and type and the amount of precomplex incorporated. Selectivity experiments were conducted in respect to isoelectric point as well as size of the competitor proteins under both uncompetitive and competitive conditions. The relative selectivity coefficients of the specific filtration cartridge in respect to isoelectronic points for interferon/ IgG, interferon/HSA and interferon/insulin pairs were 3.72, 7.10 and 10.68 times greater than the coefficient of a non-imprinted [P(HEMATrp)] filtration cartridge, respectively. Similarly, the relative selectivity coefficients of the specific filtration cartridge in respect to competitor size for interferon/ lysozyme, interferon/myoglobin and interferon/carbonic anhydrase pairs were calculated as 7.58, 10.40 and 11.68 under uncompetitive conditions whereas those values under competitive conditions were calculated as 1.08, 1.05 and 1.34, respectively. The results indicated that specific filtration cartridges developed could repeatedly adsorb interferon alpha-2b with a short separation time without any significant decrease in the adsorption capacity even if competitive conditions were conducted.

WOS: 000347580500103, PubMed ID: 25454659, Molecular imprinting is a polymerization technique that provides synthetic analogs for template molecules. Molecularly imprinted polymers (MIPs) have gained much attention due to their unique properties such as selectivity and specificity for target molecules. In this study, we focused on the development of polymeric materials with molecular recognition ability, so molecular imprinting was combined with miniemulsion polymerization to synthesize self-orienting nanoparticles through the use of an epitope imprinting approach. Thus, L-lysine imprinted nanoparticles (LMIP) were synthesized via miniemulsion polymerization technique. Immunoglobulin G (IgG) was then bound to the cavities that specifically formed for L-lysine molecules that are typically found at the C-terminus of the Fc region of antibody molecules. The resulting nanoparticles makes it possible to minimize the nonspecific interaction between monomer and template molecules. In addition, the orientation of the entire IgG molecule was controlled, and random imprinting of the IgG was prevented. The optimum conditions were determined for IgG recognition using the imprinted nanoparticles. The selectivity of the nanoparticles against IgG molecules was also evaluated using albumin and hemoglobin as competitor molecules. In order to show the self-orientation capability of imprinted nanoparticles, human serum albumin (HSA) adsorption onto both the plain nanoparticles and immobilized nanoparticles by anti-human serum albumin antibody (anti-HSA antibody) was also carried out. Due to anti-HSA antibody immobilization on the imprinted nanoparticles, the adsorption capability of nanoparticles against HSA molecules vigorously enhanced. It is proved that the oriented immobilization of antibodies was appropriately succeeded

In this study cryogels based 2-hydroxyethyl methacrylate (HEMA) functionalized with N-methacryloyl-L-histidine methyl ester (MAH) were synthesized and used for the adsorption and separation of bovine IgG. Two series of cryogels functionalized with 5 and 10 mg of MAH as pseudobioaffinity ligand were prepared and characterized by swelling test, FTIR and SEM analysis. The adsorption efficiency of the bovine immunoglobulin into cryogels is discussed with respect to the following chromatographic parameters: pH, flow rate, initial IgG concentration, adsorption time and ionic strength. Our results show good adsorption of bovine immunoglobulin under mild separation conditions at pH 7.4. The maximum binding capacity was determined (32.4 mg/g of cryogel) and demonstrates the efficiency of the used cryogels. This efficacy is clearly seen upon increasing the maximum binding capacity from 23.2 mg (obtained with cryogels with 5 mg MAH) to 32.4 mg/g (for cryogel with 10 mg MAH ligand concentration). The purity of separated fractions was evaluated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Together our observations highlights poly (HEMA-MAH) as an efficient adsorbent for bovine immunoglobulins G separation.

WOS: 000328593100001, PubMed ID: 23928053, In this study, we aimed to develop an alternative matrix able to deplete the albumin (Alb) and immunoglobulin G (IgG) from blood plasma simultaneously to prepare plasma samples for large-scale applications of blood-related proteomics. As a first step, nano-protein A nanoparticles (nanoProA) were prepared and characterized. Subsequently, cibacron blue F3GA (CB) was immobilized onto the nanoProA's to enhance their specific affinity for Alb molecules. Finally, both nanoparticles, specifically, nanoProA and CB-nanoProA, were separately embedded into cryogel structures to combine advantages of the nanoparticles with those of the cryogels. The protein adsorption was optimized using aqueous Alb and IgG solutions separately. Subsequently, competitive protein adsorption was performed using a protein mixture prepared with Alb and IgG adhering to their plasma protein ratios. Because of the CB-immobilization, the Alb depletion performance of the cryogels increased whereas the IgG depleting performance decreased. Using the nanoProA, embedded cryogel removed 99.3% of the IgG, while using the CB-nanoProA embedded cryogel removed 97.5% of the Alb content. The simultaneous depletion performances of the cryogels for Alb and IgG were characterized using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In this study, the monolithic cryogel-based adsorbents were classified as an alternative matrix to prepare plasma samples for proteomics applications at the preparative scale

Supermacroporous cryogels were synthesized and used for the purification of immunoglobulin G (IgG) from bovine serum. Imidazole functional groups were incorporated into basic polymeric backbone to gain pseudo-specificity to cryogels by using n-vinylimidazole as comonomer. The cryogels were prepared in aqueous solution of monomers partially frozen inside plastic syringe column by cryogelation. Poly(2-hydroxyethyl methacrylate-n-vinylimidazole) [poly(HEMA-VIM)] cryogels were prepared by polymerization of water-soluble functional monomer, n-vinylimidazole, with basic monomer HEMA. The characterization methods including swelling test, Fourier transform infrared spectroscopy (FTIR), elemental analysis, and scanning electron microscopy (SEM) were performed to evaluate physical and chemical properties of cryogels prepared. Bovine IgG adsorption on plain and composite cryogels was studied with respect to different parameters such as, pH, IgG concentration, flow rate, ionic strengths, and adsorption time. The best adsorption of bovine IgG was observed at pH 7.4 up to 21.1 mg per unit mass of poly(HEMA-VIM) cryogel. Elution of IgG adsorbed from the cryogels was easily achieved with 0.1 M acetate buffer containing 1 M NaCl at pH 4.0. In order to describe the adsorption process, we applied some equilibrium and kinetic adsorption models to the data. The results best fitted to Langmuir model showing monolayer protein adsorption and surface homogeneity of cryogel. Finally, we evaluated IgG purification from bovine serum under optimal condition determined. The purification efficiency and IgG purity were investigated with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS–PAGE) study.

WOS: 000384571800109, In this study, a new surface imprinting technique for preparing a hypercrosslinked monolith to remove Cd(II) ions out from aqueous solutions was proposed. The monoliths were characterized by Fourier transform infrared spectroscopy, scanning electron microscopy, thermal gravimetrical analysis, surface area measurements and elemental analysis. The reversible nature of the hypercrosslinking process was proved by repeated crosslinking and denaturation cycles by using ferric ions as an oxidant and urea as a reductant, respectively meanwhile performing surface area measurements for both situations to demonstrate the variation in the surface porosity. The multipoint BET surface areas of poly(HEMA), surface ion imprinted (Cd-SII-HM) and non-imprinted (NI-HM) monoliths were determined as 269.1 m(2) g(-1), 79.1 m(2) g(-1), and 67.4 m(2) g(-1), respectively. By breaking hypercrosslinks, the micropore volume decreased from 39.7 mm3 g-1 to 11.8 mm(3) g(-1) while the cumulative pore volume decreased from 30.7 mm(3) g(-1) to 9.1 mm(3) g(-1) during urea treatment. At the first step, the affecting factors such as initial Cd(II) ion concentrations, pH and adsorption time were optimized. Then, the selectivity of the Cd-SII-HM for Cd(II) against other metal ions was evaluated not only from singular solutions but also from triple and quadruple solutions, which included Pb(II), Zn(II) and Hg(II) ions as competitors. The relative selectivity coefficients were calculated as 3.28, 15.61 and 58.55 for Cd(II)/Zn(II), Cd(II)/Pb(II) and Cd(II)/Hg(II) pairs. The results obtained indicated that the developed reversible and easy post-crosslinking method is quite applicable for producing the surface ion-imprinted polymers with high selectivity for the template ions [Cd(II)] regarding the potential competitor ions [Zn(II), Pb(II) and Hg(II)].

WOS:000298086500032 The aim of this study is to investigate in detail the feasibility of poly(2-hydroxyethyl methacrylate-N-methacryloyl-(L)-histidine methyl ester), PHEMAH membranes for purification of immunoglobulin G (IgG) from human plasma. PHEMAH membranes were prepared by photo-polymerization technique. Then, Zn2+, Ni2+, Co2+, and Cu2+ ions were chelated directly on the PHEMAH membranes. Elemental analysis assay was performed to determine the nitrogen content and polymerized MAH was calculated as 168.5 mu mol/g. The nonspecific IgG adsorption onto the plain PHEMA membranes was negligible (about 0.25 mg/mL). A remarkable increase in the IgG adsorption capacities were achieved from human plasma with PHEMAH membranes (up to 68.4 mg/mL). Further increase was observed with the metal-chelated PHEMAH membranes (up to 118 mg/mL). The metal-chelate affinity membranes allowed the one-step separation of IgG from human plasma. The binding range of metal ions for surface histidines from human plasma followed the order: Cu2+ > Ni2+ > Zn2+ > Co2+. Adsorbed IgG was eluted using 250 mM EDTA with a purity of 94.1%. IgG molecules could be repeatedly adsorbed and eluted with the metal-chelated PHEMAH membranes without noticeable loss in their IgG adsorption capacity. (C) 2011 Wiley Periodicals, Inc. J Appl Polym Sci 123: 3476-3484, 2012

Immobilized metal ion affinity chromatography (IMAC) has been used for purification of proteins. IMAC introduces a new approach for selectively interacting biomolecules on the basis of their affinities for metal ions. The separation is based on different binding abilities of the proteins to the chelated metal ions on support. Here, N-methacryloyl-(L)-histidine methyl ester (MAH) is used as the metal-chelating ligand. Poly(hydroxyethyl methacrylate) Poly(HEMA) based membranes were prepared by photo-polymerization technique. Then, Zn(2+), Ni(2+), Co(2+), and Cu(2+) ions were chelated directly on the poly(HEMA-MAH) membranes for purification of immunoglobulin G (IgG) from human plasma.

L-Lysine imprinted poly(2-hydroxyethyl methacrylate-co-N-methacryloyl-L-aspartic acid) [P(HEMA-co-MAAsp)] cryogels were synthesized and characterized with Fourier transform infrared spectroscopy, scanning electron microscopy, surface area measurements, swelling, and squeezing tests. Specific surface area for imprinted cryogel was 34.2m(2)/g while the value was 21.3m(2)/g for non-imprinted cryogel. IgG adsorption from aqueous solution was examined in continuous mode examining the factors effecting adsorption capacity such as pH, concentration, flow rate, temperature, ionic strength, and incubation time. 0.5M NaCl was used as desorption agent. The IgG adsorption capacity was determined as 55.1 mg/g for 1.0 mg/mL IgG original concentration at 25.0°C while pH and flow rate were 7.0 and 0.5 mL/min, respectively. When human serum was used as IgG source, the removal of 90.4% of crude IgG was attained for 1/20 diluted plasma sample. The imprinted cryogel was used in ten successive cycles without significant loss in adsorption capacity. The cryogel was determined to be 1.79 times more selective to IgG than albumin and 1.45 times more selective than hemoglobin. The adsorption behavior well suited to Langmuir isotherm and the kinetics followed pseudo-second-order model. Thermodynamic parameters ΔH°, ΔS° and ΔG° for this adsorption process were also calculated.

WOS: 000331596100009, PubMed ID: 24161508, Novel aspartic acid incorporated monolithic columns were prepared to efficiently affinity purify immunoglobulin G (IgG) from human plasma. The monolithic columns were synthesised in a stainless steel HPLC column (20 cm x 5 mm id) by in situ bulk polymerisation of N-methacryloyl-L-aspartic acid (MAAsp), a polymerisable derivative of L-aspartic acid, and 2-hydroxyethyl methacrylate (HEMA). Monolithic columns [poly(2-hydroxyethyl methacrylate-N-methacryloyl-L-aspartic acid) (PHEMAsp)] were characterised by swelling studies, Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM). The monolithic columns were used for IgG adsorption/desorption from aqueous solutions and human plasma. The IgG adsorption depended on the buffer type, and the maximum IgG adsorption from aqueous solution in phosphate buffer was 0.085 mg/g at pH 6.0. The monolithic columns allowed for one-step IgG purification with a negligible capacity decrease after ten adsorption-desorption cycles

Affinity adsorption technique is increasingly used for protein purification, separation and other biochemical applications. Therapeutic molecules such as antibodies, cytokines, therapeutic DNA and plasma proteins must be purified before characterization and utilization. The aim of this study was to prepare micronsized spherical polymeric beads and to investigate the extent of their human insulin adsorption capability. Monosize poly(ethylene glycol dimethacrylate-N-methacryloyl-(L)-histidine) [poly(EDMA-MAH)] beads were prepared by modified suspension copolymerization. Functional monomer (MAH) was synthesized using methacryloyl chloride and L-histidine. The beads were characterized using scanning electron microscopy (SEM), Fourier transform infrared spectroscopy, swelling test and elemental analysis. MAH incorporation into monosize polymeric beads, having an average size around 2-3 μm, was estimated as 55.3 μmol MAH/g bead. Equilibrium swelling ratios of poly(EDMA-MAH) and poly(EDMA) beads were 65% and 55%, respectively. Adsorption experiments were performed under different conditions (i.e., pH, temperature, protein concentration and ionic strength). It was found that adsorption characteristics are strongly depend on these conditions. Maximum insulin adsorption capacity was achieved as 24.7 mg insulin/g poly(EDMA-MAH) beads. Results were well fitted to the Langmuir isotherm model. Compared with poly(EDMA-MAH), nonspecific insulin adsorption onto poly(EDMA) beads was very low (0.61 mg insulin/g bead) and can be negligible. It was observed that insulin could be repeatedly adsorbed and desorbed (at least 10 times) without significant loss in adsorption capacity.