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

WOS: 000333246400034PubMed ID: 24233543Supermacroporous 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 inter-connected pores of 10-100 mu m 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

Development of amino functionalized carbon coated magnetic nanoparticles and their application to electrochemical detection of hybridization of nucleic acids

WOS: 000393244400026PubMed ID: 28107914In our study, the development of amino functionalized carbon coated magnetic nanoparticles (NH2-CC-MNPS) and their usage for electrochemical detection of hybridization of nucleic acids have been aimed. Firstly, NH2CC-MNPS were prepared by coating of pristine Fe3O4 nanoparticles with two layers via caramelization and silanization processes respectively. After the morphological characterization with scanning electron microscopy (SEM) it was seen that NH2-CC-MNPs was spherical shaped and in 28 nm sized. Investigation of chemical composition with the help of scanning electron microscopy/energy dispersive X-ray spectroscopy (SEM/EDX) and fourier transform infrared spectroscopy (FTIR) was showed incorporation of carbon and APTES to the structure of NH2-CC-MNPs. Magnetic property of NH2-CC-MNPs after two layered coatings was demonstrated with electron spin resonance (ESR) technique and gfactor was calculated as 2.6. In the second part of this study, optimization studies have carried out onto the surface of NH2-CC-MNPS prepared in saltless phosphate-tween 20 buffer (PBTw) for the analysis of DNA hybridization. The thiol linked DNA probe sequence representing to the Hepatitis B virus (HBV) concentration, target DNA sequence concentration, the most productive hybridization time and the selection of the nanoparticle surfaces have been researched. The electrochemical detection of DNA hybridization was investigated using PGE in combination with differential pulse voltammetry (DPV) technique by measuring the guanine oxidation signal. The detection limit was calculated in the linear target DNA concentration range of 5-25 rho/mL and it was found to be 1.15 mu g/mL (20 pmol in 110 mu L solution). It has been intended to be more reproducible, more sensitive and faster results with developed biosensor technology.Ege University Scientific Research Project CoordinationEge University [13/FBE/010]; Turkish Academy of Sciences (TUBA)Turkish Academy of SciencesA.E acknowledges the financial support from Ege University Scientific Research Project Coordination (Project no.13/FBE/010), and she also would like to express her gratitude to the Turkish Academy of Sciences (TUBA) as the Principal member for its partial support

Polymeric amylase nanoparticles as a new semi-synthetic enzyme system for hydrolysis of starch

WOS: 000317528700009PubMed ID: 23498211alpha-Amylase (EC 3.2.1.1; alpha-D-1,4,glucan glucanohydrolase) catalyzes the hydrolysis of alpha-D-(1,4)-glucosidic linkages in starch, glycogen, and various malto-oligosaccharides, by releasing alpha-anomeric products. In this study, a novel method has been developed to prepare nanoprotein particles that carry alpha-amylase as a monomer by using a photosensitive microemulsion polymerization process. The nanostructured alpha-amylase with photosensitive features have been characterized by fluorescence spectroscopy, transmission electron microscopy (TEM) and Zeta Sizer. The fluorescence intensity of amylase nanoparticles was determined to be 658 a.u. at 610 nm and the average particle size of nanoamylase was found to be about 71.8 nm. Both free alpha-amylase and nanoparticles were used in the hydrolysis of starch under varying reaction conditions such as pH and temperature that affect enzyme activity and the results were compared to each other. Km values were 026 and 0.87 mM and V-max values were 0.36 IU mg(-1) and 22.32 IU mg(-1) for nanoenzyme and free enzyme, respectively. Then, thermal stability, storage stability and reusability were investigated and according to the results, activity was preserved 60% at 60 degrees C; 20% at 70-80 degrees C temperature values and 80% after 105 days storage. Finally after 10 cycles, the activity was preserved 90% and this novel enzymatic polymeric amylase nanoparticle has showed considerable potential as reusable catalyst. (C) 2012 Elsevier B.V. All rights reserved