TY - JOUR A1 - Knödler, Matthias A1 - Rühl, Clemens A1 - Emonts, Jessica A1 - Buyel, Johannes Felix T1 - Seasonal weather changes affect the yield and quality of recombinant proteins produced in transgenic tobacco plants in a greenhouse setting JF - Frontiers in Plant Science N2 - Transgenic plants have the potential to produce recombinant proteins on an agricultural scale, with yields of several tons per year. The cost-effectiveness of transgenic plants increases if simple cultivation facilities such as greenhouses can be used for production. In such a setting, we expressed a novel affinity ligand based on the fluorescent protein DsRed, which we used as a carrier for the linear epitope ELDKWA from the HIV-neutralizing antibody 2F5. The DsRed-2F5-epitope (DFE) fusion protein was produced in 12 consecutive batches of transgenic tobacco (Nicotiana tabacum) plants over the course of 2 years and was purified using a combination of blanching and immobilized metal-ion affinity chromatography (IMAC). The average purity after IMAC was 57 ± 26% (n = 24) in terms of total soluble protein, but the average yield of pure DFE (12 mg kg−1) showed substantial variation (± 97 mg kg−1, n = 24) which correlated with seasonal changes. Specifically, we found that temperature peaks (>28°C) and intense illuminance (>45 klx h−1) were associated with lower DFE yields after purification, reflecting the loss of the epitope-containing C-terminus in up to 90% of the product. Whereas the weather factors were of limited use to predict product yields of individual harvests conducted for each batch (spaced by 1 week), the average batch yields were well approximated by simple linear regression models using two independent variables for prediction (illuminance and plant age). Interestingly, accumulation levels determined by fluorescence analysis were not affected by weather conditions but positively correlated with plant age, suggesting that the product was still expressed at high levels, but the extreme conditions affected its stability, albeit still preserving the fluorophore function. The efficient production of intact recombinant proteins in plants may therefore require adequate climate control and shading in greenhouses or even cultivation in fully controlled indoor farms. KW - batch reproducibility KW - environmental correlation KW - fluorescent protein carrier KW - greenhouse cultivation KW - plant molecular farming Y1 - 2019 U6 - https://doi.org/10.3389/fpls.2019.01245 SN - 1664-462X (online-ressource) IS - 10 PB - Frontiers Media CY - Lausanne ER - TY - JOUR A1 - Bernau, C. R. A1 - Knödler, Matthias A1 - Emonts, Jessica A1 - Jäpel, Ronald Colin A1 - Buyel, Johannes Felix T1 - The use of predictive models to develop chromatography-based purification processes JF - Frontiers in Bioengineering and Biotechnology N2 - Chromatography is the workhorse of biopharmaceutical downstream processing because it can selectively enrich a target product while removing impurities from complex feed streams. This is achieved by exploiting differences in molecular properties, such as size, charge and hydrophobicity (alone or in different combinations). Accordingly, many parameters must be tested during process development in order to maximize product purity and recovery, including resin and ligand types, conductivity, pH, gradient profiles, and the sequence of separation operations. The number of possible experimental conditions quickly becomes unmanageable. Although the range of suitable conditions can be narrowed based on experience, the time and cost of the work remain high even when using high-throughput laboratory automation. In contrast, chromatography modeling using inexpensive, parallelized computer hardware can provide expert knowledge, predicting conditions that achieve high purity and efficient recovery. The prediction of suitable conditions in silico reduces the number of empirical tests required and provides in-depth process understanding, which is recommended by regulatory authorities. In this article, we discuss the benefits and specific challenges of chromatography modeling. We describe the experimental characterization of chromatography devices and settings prior to modeling, such as the determination of column porosity. We also consider the challenges that must be overcome when models are set up and calibrated, including the cross-validation and verification of data-driven and hybrid (combined data-driven and mechanistic) models. This review will therefore support researchers intending to establish a chromatography modeling workflow in their laboratory. KW - biopharmaceutical production process KW - Data-driven models KW - downstream processing design KW - experiment quality KW - hybrid model validation Y1 - 2022 U6 - https://doi.org/10.3389/fbioe.2022.1009102 SN - 2296-4185 (online-ressource) IS - 10 PB - Frontiers Media CY - Lausanne ER - TY - JOUR A1 - Emonts, Jessica A1 - Buyel, Johannes Felix T1 - An overview of descriptors to capture protein properties – Tools and perspectives in the context of QSAR modeling JF - Computational and Structural Biotechnology Journal N2 - Proteins are important ingredients in food and feed, they are the active components of many pharmaceutical products, and they are necessary, in the form of enzymes, for the success of many technical processes. However, production can be challenging, especially when using heterologous host cells such as bacteria to express and assemble recombinant mammalian proteins. The manufacturability of proteins can be hindered by low solubility, a tendency to aggregate, or inefficient purification. Tools such as in silico protein engineering and models that predict separation criteria can overcome these issues but usually require the complex shape and surface properties of proteins to be represented by a small number of quantitative numeric values known as descriptors, as similarly used to capture the features of small molecules. Here, we review the current status of protein descriptors, especially for application in quantitative structure activity relationship (QSAR) models. First, we describe the complexity of proteins and the properties that descriptors must accommodate. Then we introduce descriptors of shape and surface properties that quantify the global and local features of proteins. Finally, we highlight the current limitations of protein descriptors and propose strategies for the derivation of novel protein descriptors that are more informative. KW - Prediction of molecular features KW - Protein structure complexity KW - Quantitative structure activity relationship KW - Scalar parameters KW - Shape and surface properties Y1 - 2023 U6 - https://doi.org/10.1016/j.csbj.2023.05.022 SN - 2001-0370 (online-ressource) IS - 21 SP - 3234 EP - 3247 PB - Research Network of Computational and Structural Biotechnology CY - Gotenburg ER - TY - JOUR A1 - Rake, Heinrich A1 - Enning, Manfred A1 - Kurth, Johannes A1 - Schröder, Walter T1 - Automatic uncoupler completes automation at the hump JF - RGI - Railway Gazette International Y1 - 1994 SN - 0373-5346 VL - 150 IS - 6 SP - 371 EP - 374 PB - Reed Business CY - Sutton ER - TY - JOUR A1 - Rake, Heinrich A1 - Schwanhäußer, Wulf A1 - Frederich, Fritz A1 - Enning, Manfred T1 - Automatisierung von Ablaufanlagen mit dem Kupplungsroboter JF - ETR - Eisenbahntechnische Rundschau Y1 - 1993 SN - 0013-2845 IS - 4 SP - 249 EP - 254 PB - DVV Media Group CY - Hamburg ER - TY - JOUR A1 - Manfred, Enning A1 - May, Jörg A1 - Sutter, Stefan T1 - Innovation am Güterwagen am Beispiel der Automatischen Bremsprobe JF - ETR - Eisenbahntechnische Rundschau N2 - Mit der Digitalen Automatischen Kupplung beginnt ein neues Kapitel des Schienengüterverkehrs, in dem zusammengestellte Wagen sich automatisch in wenigen Minuten abfahrbereit machen, ohne dass der Mensch eingreifen muss. Eines des größten Hemmnisse der umweltfreundlichen Schiene wird dann entfallen. Notwendig ist jetzt eine Diskussion über den Umfang und die Systemgrenzen der Automatischen Bremsprobe. Y1 - 2020 SN - 0013-2845 IS - 12 SP - 14 EP - 19 PB - DVV Media Group CY - Hamburg ER - TY - JOUR A1 - Enning, Manfred A1 - Schmidt, Bernd A1 - Wilbring, Daniela T1 - Auf dem Weg zur autonomen Anschlussbedienung JF - Privatbahn-Magazin Y1 - 2019 SN - 1865-0163 IS - 3 SP - 40 EP - 43 PB - Bahn Media CY - Suhlendorf ER - TY - JOUR A1 - Enning, Manfred A1 - Pfaff, Raphael T1 - Güterwagen 4.0 - Mehr als nur technischer Fortschritt JF - Privatbahn-Magazin N2 - Lokomotiven sind dank modernster Konzepte der Antriebstechnik heute energiesparend und umweltfreundlich. Eine Ausrüstung mit Telematik und Assistenzfunktionen ist Standard. Auf der Strecke zeigt sich moderne Technik in Form elektronischer Stellwerke und Zugsicherungssysteme und in Rangier- und Abstellanlagen als EOW-Technik. Am Güterwagen hingegen ist der technische Fortschritt komplett vorbeigegangen. Auch beim modernsten Wagen (Abb. 1) ist die einzige „Automatik“-Funktion die zentral über die Hauptluftleitung (HL) versorgte und betätigte Luftbremse. Y1 - 2017 SN - 1865-0163 IS - 2 SP - 21 EP - 25 PB - Bahn Media CY - Suhlendorf ER - TY - JOUR A1 - Enning, Manfred A1 - Pfaff, Raphael T1 - Digitalisierung bringt mehr Güter auf die Schiene JF - Sonderprojekte ATM/MTZ Y1 - 2016 U6 - https://doi.org/10.1007/s41491-016-0570-7 SN - 2509-4610 VL - 21 IS - 6 (suppl.) SP - 34 EP - 37 PB - Springer Fachmedien CY - Wiesbaden ER - TY - JOUR A1 - Eichler, Fabian A1 - Balc, Nicolae A1 - Bremen, Sebastian A1 - Nink, Philipp T1 - Investigation of laser powder bed fusion parameters with respect to their influence on the thermal conductivity of 316L samples JF - Journal of Manufacturing and Materials Processing N2 - The thermal conductivity of components manufactured using Laser Powder Bed Fusion (LPBF), also called Selective Laser Melting (SLM), plays an important role in their processing. Not only does a reduced thermal conductivity cause residual stresses during the process, but it also makes subsequent processes such as the welding of LPBF components more difficult. This article uses 316L stainless steel samples to investigate whether and to what extent the thermal conductivity of specimens can be influenced by different LPBF parameters. To this end, samples are set up using different parameters, orientations, and powder conditions and measured by a heat flow meter using stationary analysis. The heat flow meter set-up used in this study achieves good reproducibility and high measurement accuracy, so that comparative measurements between the various LPBF influencing factors to be tested are possible. In summary, the series of measurements show that the residual porosity of the components has the greatest influence on conductivity. The degradation of the powder due to increased recycling also appears to be detectable. The build-up direction shows no detectable effect in the measurement series. KW - Additive manufacturing KW - LPBF KW - SLM KW - Thermal conductivity KW - 316L Y1 - 2024 U6 - https://doi.org/10.3390/jmmp8040166 SN - 2504-4494 N1 - Corresponding author: Fabian Eichler VL - 8 IS - 4 PB - MDPI CY - Basel ER -