Protein A Baskılanmış Süpermakrogözenekli Poli (Hidroksietil-Metakrilat) Kriyojeller

Protein A can be combined with Fc receptors of certain immunoglobulin (IgG1, IgG2, IgG4), and thus can be antifagositer and anticomplementer activity. In addition, this protein was used as a nonspecific carrier with S. aureus coaglutination constitutes the basis of tests. Protein A is the most important ligand of IgG purification, due to the connection of specific IgG. Therefore Protein A is great importance commercially.Molecular imprinting is a technology to create recognition sites in a macromolecular matrix using a template molecule. Molecularly imprinted polymers (MIP) are easy to prepare, stable, inexpensive and capable of molecular recognition. MIPs can be considered as affinity separation media.Cryogels are gel matrices that are formed in moderately frozen solutions with radicalic polymerization. The unique structure of cryogels, in combination with their osmotic, chemical and mechanical stability, makes them attractive matrices forchromatography of biological nanoparticlesProtein A bazı immunglobulinlerin (IgG1, IgG2, IgG4) Fc reseptörleri ile birleşebilmekte, böylelikle antifagositer ve antikomplementer etkinlik gösterebilmektedir. Ayrıca bu protein S. aureus un nonspesifik taşıyıcı olarak kullanıldığı koaglutinasyon testlerinin esasını oluşturmaktadır. Protein A IgG ye spesifik olarak bağlanması nedeniyle IgG saflaştırılmasında kullanılan en önemli liganttır. Bu nedenle protein A ticari olarak büyük bir öneme sahiptir.Moleküler baskılama, moleküler tanıma temelinde çok çeşitli maddelerin seçici olarak bağlanması temeline dayanan bir yöntemdir. Moleküler baskılanmış polimerler hazırlanması kolay, dayanıklı, ucuz ve moleküler tanıma yeteneği olan malzemelerdir. Moleküler baskılanmış polimerler afinite ayırma araçları olarak düşünülebilir.Kriyojeller, kısmen donmuş ortamda radikalik polimerizasyon ile hazırlanmaktadır. Kriyojellerin ozmotik, kimyasal ve mekanik kararlılık ile eşsiz yapıları birleştiğindebiyolojik nanopartiküller (plazmidler, virüsle

Preparation of Staphylococcal Protein A Imprinted Supermacroporous Cryogel Beads

Protein A is the most commonly used ligand in IgG purification due to its specific binding to the Fc receptor of most immunoglobulins, making it commercially important. Molecular imprinting is a method based on the selective recognition of various molecules. Molecular imprinted polymers are materials that are easy to prepare, durable, cheap and have molecular recognition capability. Cryogels are prepared by radical polymerization in a partially frozen environment. The unique structure of cryogels combined with osmotic, chemical and mechanical stability make them attractive chromatography matrices for a variety of biological compounds/specimens (plasmids, pathogens, cells). In this protocol, protein A imprinted supermacroporous poly(2-hydroxyethyl methacrylate) cryogels were prepared in spherical form for protein A purification. The characterization of the prepared cryogels were made by swelling test, scanning electron microscopy (SEM), Fourier transform infrared spectrophotometer (FTIR), and Brunauer–Emmett–Teller (BET) surface area analysis. After characterization, optimum conditions for protein A adsorption were determined in the batch system. The maximum protein A adsorption capacity was determined after optimization of the imprinted cryogels. Protein A relative selectivity coefficients of imprinted cryogels were examined for both Fc and protein G. Protein A was isolated from the bacterial cell wall using fast performance liquid chromatography (FPLC). The separated protein A was determined by sodium dodecyl sulfate gel electrophoresis (SDS-PAGE). In the last stage, the reusability of the cryogel was examined

Combined protein A imprinting and cryogelation for production of spherical affinity material

Cryogels have been demonstrated to be efficient when applied for protein isolation. Owing to their macroporous structure, cryogels can also be used for treating particle-containing material, e.g. cell homogenates. Another challenging development in protein purification technology is the use of molecularly imprinted polymers (MIPs). These MIPs are robust and can be used repeatedly. The paper presents a new technology that combine the formation of cryogel beads concomitantly with making imprints of a protein. Protein A was chosen as the print molecule which was also be the target in the purification step. The present paper describes a new method to produce protein-imprinted cryogel beads. The protein-imprinted material was characterized and the separation properties were evaluated with regard to both the target protein and whole cells with target protein exposed on the cell surface. The maximum protein A adsorption was 18.1 mg/g of wet cryogel beads. The selectivity coefficient of protein A-imprinted cryogel beads for protein A was 5.44 and 12.56 times greater than for the Fc fragment of IgG and protein G, respectively

Upgrading of bio-separation and bioanalysis using synthetic polymers : Molecularly imprinted polymers (MIPs), cryogels, stimuli-responsive polymers

Bio-separation plays a crucial role in many areas. Different polymers are suitable for bio-separation and are useful for applications in applications in both science and technology. Besides biopolymers, there are a broad spectrum of synthetic polymers with tailor-made properties. The synthetic polymers are characterized by their charges, solubility, hydrophilicity/hydrophobicity, sensitivity to environmental conditions and stability. Furthermore, ongoing developments are of great interest on biodegradable polymers for the treatment of diseases. Smart polymers have gained great attention due to their unique characteristics especially emphasizing simultaneously changing their chemical and physical property upon exposure to changes in environmental conditions. In this review, methodologies applied in bio-separation using synthetic polymers are discussed and efficient candidates are focused for the construction of synthetic polymers

Recent Advances in Optical Sensing for the Detection of Microbial Contaminants

Microbial contaminants are responsible for several infectious diseases, and they have been introduced as important potential food- and water-borne risk factors. They become a global burden due to their health and safety threats. In addition, their tendency to undergo mutations that result in antimicrobial resistance makes them difficult to treat. In this respect, rapid and reliable detection of microbial contaminants carries great significance, and this research area is explored as a rich subject within a dynamic state. Optical sensing serving as analytical devices enables simple usage, low-cost, rapid, and sensitive detection with the advantage of their miniaturization. From the point of view of microbial contaminants, on-site detection plays a crucial role, and portable, easy-applicable, and effective point-of-care (POC) devices offer high specificity and sensitivity. They serve as advanced on-site detection tools and are pioneers in next-generation sensing platforms. In this review, recent trends and advances in optical sensing to detect microbial contaminants were mainly discussed. The most innovative and popular optical sensing approaches were highlighted, and different optical sensing methodologies were explained by emphasizing their advantages and limitations. Consequently, the challenges and future perspectives were considered

Protein C recognition by ion-coordinated imprinted monolithic cryogels

WOS: 000399782900021PubMed: 28195421The protein C imprinted monolithic cryogel was synthesized using 2-hydroxyethyl methacrylate by redox cryo-polymerization method. The prepared monolithic cryogel was characterized by Fourier transform infrared spectroscopy, swelling test, surface area measurements, and scanning electron microscopy. The nonimprinted cryogel was prepared as well for control. Adsorption of protein C from aqueous solutions was investigated in a continuous mode and several parameters affecting adsorption performance were optimized. The maximum protein C adsorption amount was 30.4 mg/g. The selectivity studies were performed by monolithic column studies and fast protein liquid chromatography, using hemoglobin and human serum albumin as competing proteins. The relative selectivity coefficients were 2.37 and 8.89 for hemoglobin and human serum albumin, respectively. Reusability was tested for ten consecutive adsorption-desorption cycles, and no significant change in adsorption capacity was recorded. A pseudo-second-order model was suitable to interpret kinetic data, and the Langmuir model suited the adsorption isotherms well

Molecular Imprinted Nanocomposites for Green Chemistry

Nanocomposite materials which are considered ‘green’ refer to non-toxic, biodegradable and renewable nanocomposites. The reasons of preferring green nanocomposites much more could be explained by environmental friendly, fully degradability, renewability and sustainability in all respects. Furthermore, the production of green nanocomposites should not be based on toxic chemicals. When their functions are definitely completed, they can be easily destroyed without harming the environment. The challenge with green composites arises from the difficulty of producing green nano-polymers to be applied as matrices in the construction of basic composites. Molecularly imprinted polymers (MIPs) have been extensively synthesized from various functional monomers. In green chemistry principles, elimination of toxic reagents in the analytical process, the use of reagents from a renewable source are performed. To date, there are some publications pointing out the utilization of harmless chemicals for the design of MIPs. It has been a great opportunity that a novel research area has emerged considering the combination of environmentally friendly reagents and traditional organic monomers for MIP synthesis. In this chapter, the recent advances in the field of both green synthesis and green applications by focusing the molecular imprinting technology are summarized, and the developments in green strategies are highlighted

Bacterial cellulose nanofibers for separation, drug delivery, wound dressing, and tissue engineering applications

Bacterial cellulose (BC) nanofibers with high purity, high surface/volume ratio, high biocompatibility, high porosity, high tensile strength, and high water holding capacity are emerging attractive biomaterial. BC nanofibers can be prepared simply via bacterial incubation withalow cost inadetermined shape owing to its in-situ moldability. BC nanofibers due to its structural and morphological properties have demonstrated great potential asanative form or with various modifications using polymers, micro/nanoparticles, and small molecules for diverse application areas. This review aims to discuss the applicability and efficiency of the BC nanofibers and its composites focusing on the unique characteristics of BC in the fields of separation, drug delivery, wound dressing, and tissue engineering. It overviews the recent developments about the fabrication methods of BC that was used asaseparation medium, adrug delivery vehicle, awound dressing material, andascaffold