Development of etching paste for serial number restoration on aluminum engine block.

Engine numbers, which involve information regarding the engine type, production number, and year and place of manufacture, are used for identification purposes. Comprising of unique alphanumeric characters, the engine numbers are fully or partially obliterated especially in auto theft and smuggling cases to conceal the origin, identity, and owner of vehicles. The limitations of the current restoration techniques such as the difficulty of using chemical liquid etching in vertical sites, the restrictions of magnetic and optical methods, and the applicability of several techniques like electron backscatter diffraction only in the laboratory environment prompt the development of new techniques. In view of these limitations and the importance of restoring engine numbers in criminal investigations, this unique study aimed to develop an etching paste that would restore the effaced characters on a real aluminum alloy engine block. The characters which were cold-stamped on the engine block were milled at varying depths and restoration attempts were conducted using etching pastes formed with different chemicals and materials. The analyses indicate that the etching paste formed with 200 mg of perlite, 400 mg of iron powder, and 450 µL of 20 M NaOH provided restoration to a good extent. The prevention of over-etching through the controlling of the chemical reaction and the cost-effectiveness appears to be the advantages of this technique. The success of recovery on the real engine block, the facilitation of restoration on curved surfaces, and the chance of on-site usage will likely make the etching paste a widely used tool in serial number restoration.

The effects of four different solvent vapours on the restoration of obliterated stamp markings from five different wooden surfaces.

The stamp markings on wooden surfaces, which are placed on trees and products including antiques, indicate the status of trees and involve identifying data regarding the products. Such markings are obliterated either to facilitate illegal logging or to conceal product information. Despite the wide literature on the restoration of obliterated characters on metal and polymer surfaces, the recovery of defaced characters on wooden surfaces appears to be understudied. Several reference texts in the forensic marks' examination literature suggest that water, water vapor, and alkaline solutions are useful in restoring the abraded markings on the wood. Since there does not seem to be any experimental study proving such success, this study aimed to fill this gap. This study conducted experimental research by using water, ethanol, ammonia, and chloroform to recover the scraped characters on samples obtained from walnut, beech, spruce, oak, and cedar trees. The cold-stamped characters, which were defaced at varying depths, were restored using vapor and liquid phases of four solvents. While the vapor phases of water, ethanol, and ammonia yielded good outcomes on all types of wooden surfaces, the liquid phases did not seem to be useful in the revisualization process. The response of the vapors, which varied between 62 and 220 s, depended on the type of wood. The restoration technique developed in this research offers the possibility of on-site usage, easy application, utilization of low-cost solvents, rapid recovery, and effectiveness on various wooden surfaces. Overall, the restoration methodology used in this research appears to be fruitful in retrieving identifying information on wooden samples.

Designing of ZnO nanoparticles oriented interface imprinted electrochemical sensor for fluoxetine detection.

This study represents nanoparticle-based well-oriented recognition sites via interface imprinting, followed by selective and sensitive determination of fluoxetine (FLX). Herein, FLX was firstly immobilized onto ZnO NPs, and then polymerization was carried out with MAPA, HEMA, and EGDMA on the glassy carbon electrode via photopolymerization. After the etching of ZnO with and 10 mM HCI solution, a porous structure with recognition sites for FLX was constructed onto surface. The characterization of the electrochemical sensor was accomplished by utilizing CV, EIS, ATR-FTIR AFM, and SEM analysis. The DPV was used to determine FLX in standard solution, serum sample, and tap water. The effect of FLX concentration variation was studied using the DPV in the range of 1.0 × 10-11 M to 1.0 × 10-10 M with a detection limit of 2.67 × 10-12 M. This sensor showed specific recognition toward template, and more than 90% of its original response was retained after being stored in the desiccator at R.T. for 5 days. This technique has proven to be a powerful, highly selective, and sensitive tool for the rapid detection of FLX in tap water and spike serum samples.

Upconversion nanoparticles as an immunocomplexing agent for selective detection of caspases via sandwich-like supracomplexes.

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.

Synthesis and characterization of stimuli-responsive hydrogels: evaluation of external stimuli influence on L929 fibroblast viability.

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.

A sensitive and selective electrochemical sensor based on molecularly imprinted polymer for the assay of teriflunomide.

In this work, pyrrole-histidine has been designed, synthesized and, used as a novel functional monomer to fabricate a molecularly imprinted electrochemical sensor for the selective and sensitive detection of teriflunomide (TER). The molecularly imprinted thin film of electrochemical sensor was constructed by directly electropolymerization of co-polymer of pyrrole-histidine (PyHis) with pyrrole in the presence of a template, TER, on a glassy carbon electrode (GCE). After electropolymerization, the structure and morphology of the fabricated MIP sensor were characterized by Fourier-transform infrared spectroscopy (FT-IR) and scanning electron microscopy (SEM) and its electrochemical parameters such as cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS). The poly (pyrrole-co-pyrrole-histidine) [Poly (Py-co-PyHis)]@MIP/GCE sensor have a linear TER concentration in the of 0.1-1.0 pM with a low detection limit of 11.38 fM. The present strategy for electrochemical sensor have been also showed excellent recovery in synthetic serum samples and tablet dosage form with the recoveries 97.56% and 100.35%, respectively. The developed [Poly (Py-co-PyHis)]@MIP/GCE sensor exhibited an excellent electrochemical response for TER due to the synergistic effect of conducting polymer and molecularly imprinting techniques.

The investigation of parameters affecting Ibrutinib release from chitosan/tripolyphosphate/carbon nanofiber composite microspheres.

This study described the performance of carbon nanofiber modified chitosan (CNF@CS) composite microspheres for the controlled release of the Ibrutinib (IBR) drug. The surface morphology, particle sizes, and functional group contents of the microspheres were characterized by attenuated total reflection-Fourier transform infrared spectroscopy (ATR-FTIR), scanning electron, and optical microscopy measurements. The obtained data demonstrated that the addition of CNF to the microsphere increased the encapsulation efficiency of the IBR while allowing the controlled and gradual release of the drug. In terms of the encapsulation efficiency and drug release rate, IBR@CS/TPP/CNF microspheres, achieving drug encapsulation efficiency of 83.09%, have the most suitable formulation according to the comparative studies. Furthermore, according to Korsmeyer-Peppas kinetic model, IBR release mechanism was anomalous diffusion (swelling-controlled behavior and diffusion.) because the IBR release profile was completed in 78 h under optimized conditions. Therefore, the development of CNF based chitosan microsphere is a promising approach to assure appropriate dosage, safety, and improving drug efficacy.

A spectroscopic approach for rapid and simple serial number restoration on polyamide 6 parts of firearms: The use of video spectral comparator 8000.

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.

Interface imprinted polymers with well-oriented recognition sites for selective purification of hemoglobin.

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.

Purification of Fab and Fc using papain immobilized cryogel bioreactor separator system.

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.

Determination of Ochratoxin A traces in foodstuffs: Comparison of an automated on-line two-dimensional high-performance liquid chromatography and off-line immunoaffinity-high-performance liquid chromatography system.

Automated on-line two-dimensional high-performance liquid chromatography method (2D-HPLC) is proposed to determine Ochratoxin A (OTA) in food samples as an alternative to OTA immunoaffinity column (IAC). An on-line 2D-HPLC system is designed for the analysis of OTA using an affinity-based monolithic column in the first dimension and reversed-phase C18 column in the second dimension. Initially, optimal OTA separation efficiency is determined through traditional HPLC system consisting of a P(HEMAPA) monolithic column coupled with HPLC system. Secondly, after providing optimum conditions, OTA determination was investigated through the 2D-HPLC system. According to results, 2D-HPLC system showed good linearity in the range 0.5 to 20 ng/mL with limit of detection (LOD) and limit of quantification (LOQ) values of 21.2 pg/mL and 64.3 pg/mL, respectively. The P(HEMAPA)-4 monolithic column displayed good recovery of OTA ranging from 104.34% to 107.33%. Relative standard deviations (RSD) varied in the range 0.21% to 1.31% thus indicating the efficiency of P(HEMAPA)-4 monolithic column developed for OTA.

Synthesis of hydroxyethyl-methacrylate-(L)-histidine methyl ester cryogels. Application on the separation of bovine immunoglobulin G.

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.

Rapid, efficient and selective preconcentration of benzo[a]pyrene (BaP) by molecularly imprinted composite cartridge and HPLC.

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 24.86µg/L, 1.60%, and 349.6%, respectively.

Polyglycidyl methacrylate based immunoaffinity cryogels for insulin adsorption.

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.

Synthesis of L-lysine imprinted cryogels for immunoglobulin G adsorption.

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.

Antibody purification from human plasma by metal-chelated affinity membranes.

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.

Self-oriented nanoparticles for site-selective immunoglobulin G recognition via epitope imprinting approach.

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.

Aspartic acid incorporated monolithic columns for affinity glycoprotein purification.

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 × 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.

Simultaneous depletion of immunoglobulin G and albumin from human plasma using novel monolithic cryogel columns.

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.

Monosize microbeads for pseudo-affinity adsorption of human insulin.

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.