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Immobilized metal affinity chromatography (IMAC) is a widely used technique for purifying polyhistidine-tagged recombinant proteins. However, it often has practical limitations that require complex optimizations and additional steps for purification. In this study, we introduce functionalized corundum particles as a novel, efficient, and economical method for purifying recombinant proteins in a column-free format. The corundum surface is modified with amino silane APTES, followed by EDTA dianhydride, and then loaded with nickel ions. We used the Kaiser test to monitor the modification process and ICP-MS to quantify the metal-binding capacity. To evaluate the system, we used His-tagged protein A/G (PAG) mixed with bovine serum albumin (BSA). The corundum particles exhibited a binding capacity of approximately 3 mg of protein per gram of corundum or 2.4 mg per 1 mL of corundum suspension. We also examined cytoplasm obtained from different E. coli strains as an example of a complex matrix. Varying the imidazole concentration in the loading and washing buffers showed that higher concentrations during loading improved purity. Even with sample volumes as large as one liter, we successfully isolated recombinant proteins down to a concentration of 1 µg/mL. We found higher purity levels with corundum when comparing the corundum material to standard Ni–NTA agarose beads. We successfully purified His6-MBP-mSA2, a fusion protein comprising monomeric streptavidin and maltose-binding protein, from E. coli cytoplasm, demonstrating the method's applicability. We also purified SARS-CoV-2-S-RBD-His8 expressed in human Expi293F cells, confirming its suitability for mammalian cell culture supernatants. The material cost of the nickel-loaded corundum material (without regeneration) is estimated to be less than 30 cents per gram of functionalized support or 10 cents per milligram of isolated protein. Another advantage of this system is the exceptional physical and chemical stability of corundum particles. Overall, we have demonstrated that this novel material offers an efficient, robust, and cost-effective purification platform for His-tagged proteins, even in challenging, complex matrices and large sample volumes with low product concentrations. This method has potential applications in both small laboratories and large-scale industrial settings.
Industrial and agricultural waste streams (waste water, sludges, tailings, etc.) which contain high concentrations of NH4+, PO43–, and transition metals are environmentally harmful and toxic pollutants. At the same time, phosphorous and transition metals constitute highly valuable resources. Typically, separate pathways have been considered to extract hazardous transition metals or phosphate independently from each other. Investigations on the simultaneous removal of multiple components have been carried out only to a limited extent. Here, we report the synthesis routes for Ni- and Co-struvites (NH4MPO4·6H2O, M = Ni2+ and Co2+), which allow for P, ammonia, and metal co-precipitation. By evaluating different reaction parameters, the phase and stability of transition metal struvites as well as their crystal morphologies and sizes could be optimized. Ni-struvite is stable in a wide reactant concentration range and at different metal/phosphorus (M/P) ratios, whereas Co-struvite only forms at low M/P ratios. Detailed investigations of the precipitation process using ex situ and in situ techniques provided insights into the crystallization mechanisms/crystal engineering of these materials. M-struvites crystallize via intermediate colloidal amorphous nanophases, which subsequently aggregate and condense to final crystals after extended reaction times. However, the exact reaction kinetics of the formation of a final crystalline product varies significantly depending on the involved metal cation in the precipitation process: several seconds (Mg) to minutes (Ni) to hours (Co). The achieved level of control over the morphology and size makes precipitation of transition metal struvites a promising method for direct metal recovery and binding them in the form of valuable phosphate raw materials. Under this paradigm, the crystals can be potentially up-cycled as precursor powders for electrochemical or (electro)catalytic applications, which require transition metal phosphates.
Duplex stainless steels combine the positive properties of its two phases, austenite and ferrite. Due to its good corrosion resistance, high tensile strength, and good ductility, it has multiple applications. But laser beam welding of duplex steels changes the balanced phase distribution in favor of ferrite. This results in a higher vulnerability to corrosion and a lower ductility. Herein, different powder combinations consisting of duplex and nickel for coating layers by laser metal deposition (LMD) are investigated. Afterward, laser tracks are welded, and the temperature cycles are measured. The ferrite content of the tracks is analyzed by feritscope, metallographic analysis, and electron backscatter diffraction. The goal is the development of a powder mixture allowing for a duplex microstructure in a two-step process, where first the edges of the weld partners are coated with the powder mixture by LMD and second those edges are laser beam welded. The powder mixture identified by the pretests is tested in the two-step process and analyzed by metallographic analysis, energy-dispersive X-ray spectroscopy, and
Vickers hardness tests. The resulting weld seams show a balanced duplex microstructure with a homogenous nickel distribution and a hardness of the weld seam similar to the base material.
Duplex stainless steels combine the positive properties of its two phases, austenite and ferrite. Due to its good corrosion resistance, high tensile strength and good ductility it has multiple applications. But laser beam welding of duplex steels changes the balanced phase distribution in favor of ferrite. This results in a higher vulnerability to corrosion and a lower ductility. In this study different powder combinations consisting of duplex and nickel for coating layers by laser metal deposition are investigated. Afterwards laser tracks are welded, and the temperature cycles measured. The ferrite content of the tracks are analyzed by feritscope, metallographic analysis and Electron Backscatter Diffraction. The goal is the development of a powder mixture allowing for a duplex microstructure in a two-step process, where firstly the edges of the weld partners are coated with the powder mixture by LMD and secondly those edges are laser beam welded. The powder mixture identified by the pretests is tested in the two-step process and analyzed by metallographic analysis, energy dispersive X-ray spectroscopy and Vickers hardness tests. The resulting weld seams show a balanced duplex microstructure with a homogenous nickel distribution and a hardness of the weld seam similar to the base material.
The effects of rapid pulse electric current sintering (PECS), substitution of WC by NbC and Co by Ni, and carbide additives (TiC and Mo2C) on the microstructure, elastic modulus, B3B transverse rupture strength (TRS) and high temperature sliding wear on WC-Co, WC-Ni, NbC-Co and NbC-Ni cermets were studied. High temperature sliding wear tests were carried out using a ball-on-disk tribometer, with a 10 N force, at a sliding speed of 1.34 m/s for 0.8 km (10 min) and 2.4 km (30 min), using 100Cr6 (AISI 52100) steel balls at 400 °C and 0% humidity. Additions of TiC and Mo2C to NbC-12Ni improved the sliding wear resistance, with TiC having the greater effect, reducing the sample wear rate by over 30% from 15.1×10−6mm3/N·m to 9.4×10−6mm3/N·m after sliding distance of 2.4 km. Generally, the LPS samples had lower wear volumes than the corresponding SPS samples, due to higher K1c and TRS.
The movement of the macroscopic magnetic moment in ferromagnetic systems can be described by the Landau-Lifshitz (LL) or Landau-Lifshitz-Gilbert (LLG) equation. These equations are strictly valid only at absolute zero temperature. To include temperature effects a stochastic version of the LL or LLG equation for a spin density of one per unit cell can be used instead. To apply the stochastic LL to micromagnetic simulations, where the spin density per unit cell is generally higher, a conversion regarding simulation cell size and temperature has to be established. Based on energetic considerations, a conversion for ferromagnetic bulk and thin film systems is proposed. The conversion is tested in micromagnetic simulations which are performed with the Object Oriented Micromagnetic Framework (OOMMF). The Curie temperatures of bulk Nickel, Cobalt and Iron systems as well as Nickel thin-film systems with thicknesses between 6.3 mono layer (ML) and 31ML are determined from micromagnetic simulations. The results show a good agreement with experimentally determined Curie temperatures of bulk and thin film systems when temperature scaling is performed according to the presented model.
Research and development of WC grades is 90 years ahead of niobium carbide-based hard materials, which showed already as light-weight and unexplored material a very high potential for many technical applications, especially for wear protection and machining. NbC evoluted recently from lab scale to pilot scale, especially in areas where established WC-based materials are causing economic, environmental or technical concerns. Benchmark trials of NbC-based hard metals indicated already a favorable performance in machining of steel components.
The present work deals with Niobcarbid (NbC). Strategic reflections on tungsten carbide and more and more stringent toxicological restrictions for cobalt associated for both with spiraling stock market prices have attracted recently some attention for Niobium carbide as a substitute for tungsten carbide in machining.
The present work deals with Niobcarbid (NbC). Strategic reflections on tungsten carbide and more and more stringent toxicological restrictions for cobalt associated for both with spiraling stock market prices have attracted recently some attention for Niobium carbide as a substitute for tungsten carbide in machining.