TY - JOUR A1 - Völzke, Jule L. A1 - Smatty, Sarah A1 - Döring, Sarah A1 - Ewald, Shireen A1 - Oelze, Marcus A1 - Fratzke, Franziska A1 - Flemig, Sabine A1 - Konthur, Zoltán A1 - Weller, Michael G. T1 - Efficient Purification of Polyhistidine-Tagged Recombinant Proteins Using Functionalized Corundum Particles N2 - 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. N2 - Die immobilisierte Metallaffinitätschromatographie (IMAC) ist eine weit verbreitete Technik zur Reinigung von rekombinanten Proteinen mit Polyhistidin-Markierung. Sie hat jedoch oft praktische Einschränkungen, die komplexe Optimierungen und zusätzliche Schritte für die Aufreinigung erfordern. In dieser Studie stellen wir funktionalisierte Korundpartikel als neuartige, effiziente und wirtschaftliche Methode zur Reinigung rekombinanter Proteine in einem säulenfreien Format vor. Die Korundoberfläche wird mit dem Aminosilan APTES und anschließend mit EDTA-Dianhydrid modifiziert und dann mit Nickelionen beladen. Wir haben den Kaiser-Test zur Überwachung des Modifizierungsprozesses und ICP-MS zur Quantifizierung der Metallbindungskapazität verwendet. Zur Charakterisierung des Systems verwendeten wir His-markiertes Protein A/G (PAG) in Kombination mit Rinderserumalbumin (BSA). Die Korundpartikel wiesen eine Bindungskapazität von etwa 3 mg Protein pro Gramm Korund oder 2,4 mg pro 1 ml Korundsuspension auf. Als Beispiel für eine komplexe Matrix untersuchten wir auch Zytoplasma, das aus verschiedenen E. coli-Stämmen gewonnen wurde. Die Variation der Imidazolkonzentration in den Lade- und Waschpuffern zeigte, dass höhere Konzentrationen beim Laden die Reinheit verbesserten. Selbst bei einem Probenvolumen von bis zu einem Liter konnten wir rekombinante Proteine von 1 µg/mL isolieren. Beim Vergleich des Korundmaterials mit Standard-Ni-NTA-Agarose-Beads stellten wir einen höheren Reinheitsgrad mit Korund fest. Wir reinigten erfolgreich His6-MBP-mSA2 aus E. coli-Zytoplasma, ein Fusionsprotein, das aus monomerem Streptavidin und Maltose-bindendem Protein besteht. Wir reinigten auch SARS-CoV-2-S-RBD-His8, das in humanen Expi293F-Zellen exprimiert wurde, und bestätigten damit die Eignung des Materials für Zellkulturüberstände von Säugetieren. Die Materialkosten für nickelbeladenen Korund (ohne Regenerierung) werden auf weniger als 30 Cent pro Gramm funktionalisierten Trägers oder 10 Cent pro Milligramm isolierten Proteins geschätzt. Ein weiterer Vorteil dieses Systems ist die außergewöhnliche physikalische und chemische Stabilität der Korundpartikel. Insgesamt haben wir gezeigt, dass dieses neuartige Material eine effiziente, robuste und kostengünstige Reinigungsplattform für His-markierte Proteine bietet, selbst bei schwierigen, komplexen Matrices und großen Probenmengen mit niedrigen Produktkonzentrationen. Diese Methode könnte sowohl in kleinen Labors als auch in der Großindustrie eingesetzt werden. KW - Aluminum oxide KW - Sapphire KW - Ethylenediaminetetraacetic acid KW - Nickel chelate KW - EDTAD KW - HexaHis-Tag KW - His6 KW - 6xHis KW - His8 KW - Bioseparation KW - IMAC purification KW - Immunocapture KW - Affinity chromatography KW - Carrier KW - Nickel KW - Recombinant protein KW - Escherichia coli KW - Bacterial lysates KW - Cytoplasm KW - Polishing KW - Downstream processing PY - 2023 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:b43-575672 VL - 12 IS - 2 SP - 1 EP - 18 PB - MDPI CY - Basel, Schweiz AN - OPUS4-57567 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Karafiludis, Stephanos A1 - de Oliveira Guilherme Buzanich, Ana A1 - Kochovski, Z. A1 - Feldmann, Ines A1 - Emmerling, Franziska A1 - Stawski, Tomasz T1 - Ni- and Co-Struvites: Revealing Crystallization Mechanisms and Crystal Engineering toward Applicational Use of Transition Metal Phosphates N2 - 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. KW - Crystallization KW - Struvite KW - Nickel KW - Cobalt KW - Phosphorous recovery KW - Up-cycling KW - Aqueous synthesis PY - 2022 U6 - https://doi.org/10.1021/acs.cgd.2c00284 VL - 22 IS - 7 SP - 4305 EP - 4315 PB - ACS Publications CY - Washington D.C. AN - OPUS4-55286 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Straße, Anne A1 - Gumenyuk, Andrey A1 - Rethmeier, Michael T1 - Study on Duplex Stainless Steel Powder Compositions for the Coating of thick Plates of Laser Beam Welding N2 - 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. KW - Duplex AISI 2205 KW - Laser metal deposition KW - Laser beam welding KW - Nickel KW - Stainless steels PY - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:b43-547024 SN - 1438-1656 SP - 1 EP - 11 PB - Wiley-VCH CY - Weinheim AN - OPUS4-54702 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Straße, Anne A1 - Gumenyuk, Andrey A1 - Rethmeier, Michael T1 - Study on Duplex Stainless Steel Powder Compositions forthe Coating of Thick Plates for Laser Beam Welding N2 - 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. KW - Duplex AISI 2205 KW - Stainless Steel KW - Laser Beam Welding KW - Nickel KW - Laser Metal Deposition PY - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:b43-554663 SN - 1438-1656 VL - 24 IS - 6 SP - 1 EP - 11 PB - Wiley-VCH CY - Weinheim AN - OPUS4-55466 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Genga, R. M. A1 - Rokebrand, P. A1 - Cornish, L. A. A1 - Nelwalani, N. A1 - Brandt, Guido A1 - Kelling, Norbert A1 - Woydt, Mathias A1 - Janse van Vuuren, A. A1 - Polese, C. T1 - High-temperature sliding wear, elastic modulus and transverse rupture strength of Ni bonded NbC and WC cermets N2 - 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. KW - Pulse electric current sintering KW - Niobium carbide KW - Nickel KW - Molybdenum carbide KW - Titanium carbide PY - 2020 U6 - https://doi.org/10.1016/j.ijrmhm.2019.105143 SN - 0263-4368 VL - 87 SP - 105143 PB - Elsevier Ltd. AN - OPUS4-51473 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Hahn, Marc Benjamin T1 - Temperature in micromagnetism: Cell size and scaling effects of the stochastic Landau-Lifshitz equation N2 - 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. KW - Micromagnetism KW - LLG KW - LL equation KW - Landau Lifshitz equation KW - Landau Lifshitz Gilbert equation KW - Stochastic Landau Lifshitz equation KW - Stochastic Landau Lifshitz Gilbert equation KW - Curie temperature KW - Magnetic Nanoparticles KW - Thin film systems KW - Temeprature scaling KW - Phase transition KW - Magnet coupling KW - Ferromagnetism KW - Superparamagnetism KW - Paramagnetism KW - Ni KW - Co KW - Fe KW - Steel KW - Nickel KW - Cobalt KW - Iron KW - Temperature effects KW - Cell size KW - Damping factor KW - Gamma KW - Alpha KW - Spin KW - Magnetic moment KW - Magnetic interacion KW - Magnetization dynamics KW - Domain wall KW - Exchange length KW - temeprature dependent exchange length KW - Bloch wall KW - Neel wall KW - Exchange interaction KW - Magnetic anisotropy KW - Simulation KW - OOMMF KW - Object oriented micromagnetic framework PY - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:b43-484610 VL - 3 IS - 7 SP - 075009-1 EP - 075009-8 PB - IOPscience CY - England AN - OPUS4-48461 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Woydt, Mathias A1 - Huang, S. A1 - Vleugels, J. A1 - Mohrbacher, H. A1 - Cannizza, E. T1 - Potentials of niobium carbide (NbC) as cutting tools and for wear protection N2 - 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. KW - Niobium carbide KW - Friction KW - Wear KW - NbC KW - Hard metals KW - Machining KW - Nickel PY - 2018 U6 - https://doi.org/10.1016/j.ijrmhm.2018.01.009 SN - 0263-4368 VL - 72 SP - 380 EP - 387 PB - Elsevier Ltd. AN - OPUS4-44166 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Woydt, Mathias T1 - Machining with niobium carbide based tools N2 - 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. T2 - 59. Tribologie-Fachtagung CY - Goettingen, Germany DA - 24.09.2018 KW - Hard tungsten KW - Semi-finishing KW - Niobium carbide KW - Friction KW - Wear KW - NbC KW - Hard metal KW - Cobalt KW - Nickel KW - Fe3Al PY - 2018 SN - 978-3-9817451-3-9 SP - 48/1 EP - 48/11 AN - OPUS4-46170 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Woydt, Mathias T1 - Tailoring the functional profile of niobium carbide (NbC) as cutting tool materials and for wear protection N2 - 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. T2 - WORLDPM2018 CY - Beijing, China DA - 16.09.2018 KW - Niobium carbide KW - NbC KW - Machining KW - Hard turning KW - Cobalt KW - Nickel PY - 2018 AN - OPUS4-46198 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Vleugels, J. A1 - Cannizza, E. A1 - Woydt, Mathias A1 - Mohrbacher, H. T1 - Influence of sintering temperature and carbide additions on the microstructure and mechanical properties of NbC-Ni based cermets N2 - This study reports on the effect of the sintering temperature and VC, Mo2C and TiC as well as the combination of Mo+VC on the densification, NbC grain growth, microstructure evolution as well as mechanical properties of novel NbC-Ni based cermets. KW - Cermet KW - Niobium carbide KW - Nickel KW - Sintering KW - Mechanical properties PY - 2018 U6 - https://doi.org/10.3969/j.issn.1003-7292.2018.02.001 SN - 1003-7292 VL - 35 IS - 2 SP - 69 EP - 78 PB - Yingzhi Hejin Bianjibu CY - Zhuzhou AN - OPUS4-46136 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -