@misc{BrinkmannOberrathAwakowiczetal., author = {Brinkmann, Ralf Peter and Oberrath, Jens and Awakowicz, Peter and Lapke, Martin and Musch, Thomas and Mussenbrock, Thomas and Rolfes, Ilona and Schulz, Christian and Storch, Robert and Styrnoll, Tim and Zietz, Christian}, title = {Device and Use of the Device for Measuring the Density and/or the Electron Temperature and/or the Collision Frequency of a Plasma}, language = {en} } @misc{SchulzSchneiderKohlbecheretal., author = {Schulz, Christian M. and Schneider, Erich and Kohlbecher, Stefan and Hapfelmeier, Alexander and Heuser, Fabian}, title = {The influence of anaesthetists' experience on workload, performance and visual attention during simulated critical incidents}, series = {Journal of Clinical Monitoring and Computing}, volume = {28}, journal = {Journal of Clinical Monitoring and Computing}, number = {5}, issn = {1387-1307}, doi = {10.1007/s10877-013-9443-8}, pages = {475 -- 480}, language = {en} } @misc{LehnenHeuserSağlametal., author = {Lehnen, Nadine and Heuser, Fabian and Sağlam, Murat and Schulz, Christian M. and Wagner, Klaus J. and Taki, Masakatsu and Kochs, Eberhard F. and Jahn, Klaus and Schneider, Erich}, title = {Opioid-induced nausea involves a vestibular problem preventable by head-rest}, series = {PLoS one}, volume = {10}, journal = {PLoS one}, number = {8}, issn = {1932-6203}, doi = {10.1371/journal.pone.0135263}, pages = {e0135263}, language = {en} } @misc{SchulzSkrzypczakSchneideretal., author = {Schulz, Christian M. and Skrzypczak, M. and Schneider, Erich and Hapfelmeier, Alexander and Martin, J. and Kochs, Eberhard F. and Schneider, G.}, title = {Assessment of subjective workload in an anaesthesia simulator environment: reliability and validity}, series = {European Journal of Anaesthesiology}, volume = {28}, journal = {European Journal of Anaesthesiology}, number = {7}, issn = {0265-0215}, pages = {502 -- 505}, language = {en} } @misc{HeuserSchulzSağlametal., author = {Heuser, Fabian and Schulz, Christian and Sağlam, Murat and Ramaioli, Cecilia and Heuberger, Maria and Wagner, Klaus J. and Jahn, Klaus and Schneider, Erich and Brandt, Thomas and Glasauer, Stefan and Lehnen, Nadine}, title = {Preventing opioid-induced nausea and vomiting: Rest your head and close your eyes?}, series = {PloS one}, volume = {12}, journal = {PloS one}, number = {3}, issn = {1932-6203}, doi = {10.1371/journal.pone.0173925}, pages = {e0173925}, language = {en} } @incollection{SchmidtSchulz, author = {Schmidt, Sindy and Schulz, Christian}, title = {Entwicklung eines Steuerungs- und Regelungsprogrammes f{\"u}r einen Pr{\"u}fstand zur Pr{\"u}fung von Feuerwehrpumpen}, series = {Virtuelle Instrumente in der Praxis 2015 : Begleitband zum 20. VIP-Kongress}, booktitle = {Virtuelle Instrumente in der Praxis 2015 : Begleitband zum 20. VIP-Kongress}, publisher = {VDE Verlag GmbH}, address = {Berlin}, isbn = {978-3-8007-3669-0}, pages = {79 -- 81}, abstract = {Kurzdokumentation {\"u}ber die Programmierung einer Software, f{\"u}r die Anwendung automatischer Pr{\"u}froutinen, zur Pr{\"u}fung von Feuerwehrpumpen, Armaturen und Saugschl{\"a}uchen, unter Verwendung von LabVIEW als grafische Programmieroberfl{\"a}che.}, language = {de} } @misc{PrillSinghSeeberetal., author = {Prill, Robert and Singh, Jasvinder A. and Seeber, Gesine H. and Mai Nielsen, Sabrina and Goodman, Susan and Michel, Sven and Kopkow, Christian and Schulz, Robert and Choong, Peter and Hommel, Hagen}, title = {Patient, physiotherapist and surgeon endorsement of the core domain set for total hip and total knee replacement in Germany: a study protocol for an OMERACT initiative}, series = {BMJ open}, volume = {10}, journal = {BMJ open}, number = {6}, issn = {2044-6055}, doi = {10.1136/bmjopen-2019-035207}, pages = {8}, language = {en} } @misc{SchulzKruegerGengeLendleinetal., author = {Schulz, Christian and Kr{\"u}ger-Genge, Anne and Lendlein, Andreas and K{\"u}pper, Jan-Heiner and Jung, Friedrich}, title = {Potential Effects of Nonadherent on Adherent Human Umbilical Venous Endothelial Cells in Cell Culture}, series = {International Journal of Molecular Science}, volume = {22}, journal = {International Journal of Molecular Science}, number = {3}, doi = {10.3390/ijms22031493}, pages = {15}, abstract = {The adherence and shear-resistance of human umbilical venous endothelial cells (HUVEC) on polymers is determined in vitro in order to qualify cardiovascular implant materials. In these tests, variable fractions of HUVEC do not adhere to the material but remain suspended in the culture medium. Nonadherent HUVEC usually stop growing, rapidly lose their viability and can release mediators able to influence the growth and function of the adherent HUVEC. The aim of this study was the investigation of the time dependent behaviour of HUVEC under controlled nonadherent conditions, in order to gain insights into potential influences of these cells on their surrounding environment in particular adherent HUVEC in the context of in vitro biofunctionality assessment of cardiovascular implant materials. Data from adherent or nonadherent HUVEC growing on polystyrene-based cell adhesive tissue culture plates (TCP) or nonadhesive low attachment plates (LAP) allow to calculate the number of mediators released into the culture medium either from adherent or nonadherent cells. Thus, the source of the inflammatory mediators can be identified. For nonadherent HUVEC, a time-dependent aggregation without further proliferation was observed. The rate of apoptotic/dead HUVEC progressively increased over 90\% within two days. Concomitant with distinct blebbing and loss of membrane integrity over time, augmented releases of prostacyclin (PGI2, up to 2.91 ± 0.62 fg/cell) and platelet-derived growth factor BB (PDGF-BB, up to 1.46 ± 0.42 fg/cell) were detected. The study revealed that nonadherent, dying HUVEC released mediators, which can influence the surrounding microenvironment and thereby the results of in vitro biofunctionality assessment of cardiovascular implant materials. Neglecting nonadherent HUVEC bears the risk for under- or overestimation of the materials endothelialization potential, which could lead to the loss of relevant candidates or to uncertainty with regard to their suitability for cardiac applications. One approach to minimize the influence from nonadherent endothelial cells could be their removal shortly after observing initial cell adhesion. However, this would require an individual adaptation of the study design, depending on the properties of the biomaterial used.}, language = {en} } @misc{PrillSinghSeeberetal., author = {Prill, Robert and Singh, Jasvinder A. and Seeber, Gesine H. and Mai Nielsen, Sabrina and Goodman, Susan and Michel, Sven and Kopkow, Christian and Schulz, Robert and Choong, Peter and Hommel, Hagen}, title = {Endorsement des OMERACT Core Domain Sets f{\"u}r H{\"u}ft- und Kniegelenkersatz: ein Survey unter Patienten, Physiotherapeuten und Orthop{\"a}den in Deutschland - ein Studienprotokoll}, series = {4. Forschungssymposium Physiotherapie, FSPT2019 Abstractband}, journal = {4. Forschungssymposium Physiotherapie, FSPT2019 Abstractband}, pages = {45 -- 46}, language = {de} } @misc{SchulzJungKuepper, author = {Schulz, Christian and Jung, Friedrich and K{\"u}pper, Jan-Heiner}, title = {Inhibition of phase-1 biotransformation and cytostatic effects of diphenyleneiodonium on hepatoblastoma cell line HepG2 and a CYP3A4-overexpressing HepG2 cell clone}, series = {Clinical Hemorheology and Microcirculation}, volume = {79}, journal = {Clinical Hemorheology and Microcirculation}, number = {1}, issn = {1875-8622}, doi = {10.3233/CH-219117}, pages = {231 -- 243}, abstract = {Cell-based in vitro liver models are an important tool in the development and evaluation of new drugs in pharmacological and toxicological drug assessment. Hepatic microsomal enzyme complexes, consisting of cytochrome P450 oxidoreductase (CPR) and cytochrome P450 monooxygenases (CYPs), play a decisive role in catalysing phase-1 biotransformation of pharmaceuticals and xenobiotics. For a comprehensive understanding of the phase-1 biotransformation of drugs, the availability of well-characterized substances for the targeted modulation of in vitro liver models is essential. In this study, we investigated diphenyleneiodonium (DPI) for its ability to inhibit phase-1 enzyme activity and further its toxicological profile in an in vitro HepG2 cell model with and without recombinant expression of the most important drug metabolization enzyme CYP3A4. Aim of the study was to identify effective DPI concentrations for CPR/CYP activity modulation and potentially associated dose and time dependent hepatotoxic effects. The cells were treated with DPI doses up to 5,000nM (versus vehicle control) for a maximum of 48 h and subsequently examined for CYP3A4 activity as well as various toxicological relevant parameters such as cell morphology, integrity and viability, intracellular ATP level, and proliferation. Concluding, the experiments revealed a time- and concentration-dependent DPI mediated partial and complete inhibition of CYP3A4 activity in CYP3A4 overexpressing HepG2-cells (HepG2-CYP3A4). Other cell functions, including ATP synthesis and consequently the proliferation were negatively affected in both in vitro cell models. Since neither cell integrity nor cell viability were reduced, the effect of DPI in HepG2 can be assessed as cytostatic rather than cytotoxic.}, language = {en} } @misc{KruegerGengeKoehlerLaubeetal., author = {Kr{\"u}ger-Genge, Anne and K{\"o}hler, Susanne and Laube, Markus and Haileka, Vanessa and Lemm, Sandy and Majchrzak, Karolina and Kammerer, Sarah and Schulz, Christian and Storsberg, Joachim and Pietzsch, Jens and K{\"u}pper, Jan-Heiner and Jung, Friedrich}, title = {Anti-Cancer Prodrug Cyclophosphamide Exerts Thrombogenic Effects on Human Venous Endothelial Cells Independent of CYP450 Activation—Relevance to Thrombosis}, series = {Cells}, volume = {12}, journal = {Cells}, number = {15}, issn = {2073-4409}, doi = {10.3390/cells12151965}, abstract = {Cancer patients are at a very high risk of serious thrombotic events, often fatal. The causes discussed include the detachment of thrombogenic particles from tumor cells or the adverse effects of chemotherapeutic agents. Cytostatic agents can either act directly on their targets or, in the case of a prodrug approach, require metabolization for their action. Cyclophosphamide (CPA) is a widely used cytostatic drug that requires prodrug activation by cytochrome P450 enzymes (CYP) in the liver. We hypothesize that CPA could induce thrombosis in one of the following ways: (1) damage to endothelial cells (EC) after intra-endothelial metabolization; or (2) direct damage to EC without prior metabolization. In order to investigate this hypothesis, endothelial cells (HUVEC) were treated with CPA in clinically relevant concentrations for up to 8 days. HUVECs were chosen as a model representing the first place of action after intravenous CPA administration. No expression of CYP2B6, CYP3A4, CYP2C9 and CYP2C19 was found in HUVEC, but a weak expression of CYP2C18 was observed. CPA treatment of HUVEC induced DNA damage and a reduced formation of an EC monolayer and caused an increased release of prostacyclin (PGI2) and thromboxane (TXA) associated with a shift of the PGI2/TXA balance to a prothrombotic state. In an in vivo scenario, such processes would promote the risk of thrombus formation.}, language = {en} } @misc{SchulzKammererKuepper, author = {Schulz, Christian and Kammerer, Sarah and K{\"u}pper, Jan-Heiner}, title = {NADPH-cytochrome P450 reductase expression and enzymatic activity in primary-like human hepatocytes and HepG2 cells for in vitro biotransformation studies}, series = {Clinical Hemorheology and Microcirculation}, volume = {73}, journal = {Clinical Hemorheology and Microcirculation}, number = {1}, issn = {1386-0291}, doi = {10.3233/CH-199226}, pages = {249 -- 260}, language = {en} } @incollection{SchmidtHeiseSchulz, author = {Schmidt, Sindy and Heise, Philipp and Schulz, Christian}, title = {Entwicklung eines Steuerungs- und Regelungsprogrammes f{\"u}r einen Pr{\"u}fstand zur Pr{\"u}fung von Feuerwehrpumpen}, series = {Institut f{\"u}r Umwelttechnik und Recycling Senftenberg e.V. : Res{\"u}mee und ausgew{\"a}hlte Projekte 2012-2017 : 20 Jahre IURS e.V.}, booktitle = {Institut f{\"u}r Umwelttechnik und Recycling Senftenberg e.V. : Res{\"u}mee und ausgew{\"a}hlte Projekte 2012-2017 : 20 Jahre IURS e.V.}, edition = {1. Auflage}, publisher = {IURS e.V.}, address = {Senftenberg}, isbn = {978-3-00-058468-8}, pages = {86 -- 89}, abstract = {Kurzdokumentation {\"u}ber die Programmierung einer Software, f{\"u}r die Anwendung automatischer Pr{\"u}froutinen, zur Pr{\"u}fung von Feuerwehrpumpen, Armaturen und Saugschl{\"a}uchen, unter Verwendung von LabVIEW als grafische Programmieroberfl{\"a}che.}, language = {de} } @misc{SteinbrechtPfeiferHerzogetal., author = {Steinbrecht, Susanne and Pfeifer, Nadine and Herzog, Natalie and Katzenberger, Nadine and Schulz, Christian and Kammerer, Sarah and K{\"u}pper, Jan-Heiner}, title = {HepG2-1A2 C2 and C7: Lentivirus vector-mediated stable and functional overexpression of cytochrome P450 1A2 in human hepatoblastoma cells}, series = {Toxicology Letters}, volume = {319}, journal = {Toxicology Letters}, issn = {0378-4274}, doi = {10.1016/j.toxlet.2019.11.006}, pages = {155 -- 159}, abstract = {Novel HepG2 cell clones 1A2 C2 and 1A2 C7 were independently generated by lentiviral transduction to functionally overexpress cytochrome P450 1A2 (CYP1A2). We found similar and stable CYP1A2 transcript and protein levels in both cell clones leading to specific enzyme activities of about 370 pmol paracetamol x min-1 x mg-1 protein analyzed by phenacetin conversion. Both clones showed dramatically increased sensitivity to the hepatotoxic compound aflatoxin B1 (EC50<100 nM) when compared to parental HepG2 cells (EC50 ∼5 μM). Thus, newly established cell lines are an appropriate tool to study metabolism and toxicity of substances depending on conversion by CYP1A2.}, language = {en} } @misc{SchulzKruegerGengeJungetal., author = {Schulz, Christian and Kr{\"u}ger-Genge, Anne and Jung, Friedrich and Lendlein, Andreas}, title = {Aptamer supported in vitro endothelialization of poly(ether imide) films}, series = {Clinical Hemorheology and Microcirculation}, volume = {75}, journal = {Clinical Hemorheology and Microcirculation}, number = {2}, issn = {1875-8622}, doi = {10.3233/CH-190775}, pages = {201 -- 217}, abstract = {Implantation of synthetic small-diameter vascular bypass grafts is often associated with an increased risk of failure, due to thrombotic events or late intimal hyperplasia. As one of the causes an insufficient hemocompatibility of the artificial surface is discussed. Endothelialization of synthetic grafts is reported to be a promising strategy for creating a self-renewing and regulative anti-thrombotic graft surface. However, the establishment of a shear resistant cell monolayer is still challenging. In our study, cyto- and immuno-compatible poly(ether imide) (PEI) films were explored as potential biomaterial for cardiovascular applications. Recently, we reported that the initial adherence of primary human umbilical vein endothelial cells (HUVEC) was delayed on PEI-films and about 9 days were needed to establish a confluent and almost shear resistant HUVEC monolayer. To accelerate the initial adherence of HUVEC, the PEI-film surface was functionalized with an aptamer-cRGD peptide based endothelialization supporting system. With this functionalization the initial adherence as well as the shear resistance of HUVEC on PEI-films was considerable improved compared to the unmodified polymer surface. The in vitro results confirm the general applicability of aptamers for an efficient functionalization of substrate surfaces.}, language = {en} } @misc{SchulzSchneiderFritzetal., author = {Schulz, Christian M. and Schneider, Erich and Fritz, L. and Vockeroth, Johannes and Hapfelmeier, Alexander and Wasmaier, M.}, title = {Eye tracking for assessment of workload: a pilot study in an anaesthesia simulator environment}, series = {British journal of anaesthesia}, volume = {106}, journal = {British journal of anaesthesia}, number = {1}, issn = {1471-6771}, doi = {10.1093/bja/aeq307}, pages = {44 -- 50}, language = {en} } @misc{SchulzSchneiderFritzetal., author = {Schulz, Christian M. and Schneider, Erich and Fritz, L. and Vockeroth, Johannes and Hapfelmeier, Alexander and Brandt, Thomas}, title = {Visual attention of anaesthetists during simulated critical incidents}, series = {British Journal of Anaesthesia}, volume = {106}, journal = {British Journal of Anaesthesia}, number = {6}, issn = {1471-6771}, doi = {10.1093/bja/aer087}, pages = {807 -- 813}, language = {en} } @inproceedings{BauerDreyerKahleetal., author = {Bauer, Monika and Dreyer, Christian and Kahle, Olaf and Schuldt, U. and Uhlig, C. and Schulze, K. and Schulz, Stefan E. and Uhlig, M. and Geßner, Thomas}, title = {Comparison of Fluorinated and Non-Fluorinated Novel Low-k Polycyanurates for Integrated Circuit (IC) Metallization}, language = {en} } @incollection{HohbergElmerRusselletal., author = {Hohberg, Karin and Elmer, Michael and Russell, David J. and Christian, Axel and Schulz, Hans-J{\"u}rgen and Lehmitz, Ricarda and Wanner, Manfred}, title = {First five years of soil food-web development in Chicken Creek catchment}, series = {The artificial catchment Chicken Creek - initial ecosystem development 2005-2010}, booktitle = {The artificial catchment Chicken Creek - initial ecosystem development 2005-2010}, editor = {Elmer, Michael and Schaaf, Wolfgang and Biemelt, Detlef and Gerwin, Werner and H{\"u}ttl, Reinhard F.}, publisher = {FZLB}, address = {Cottbus}, pages = {93 -- 114}, language = {en} } @misc{SchulzHerzogKubicketal., author = {Schulz, Christian and Herzog, Natalie and Kubick, Stefan and Jung, Friedrich and K{\"u}pper, Jan-Heiner}, title = {Stable Chinese Hamster Ovary Suspension Cell Lines Harboring Recombinant Human Cytochrome P450 Oxidoreductase and Human Cytochrome P450 Monooxygenases as Platform for In Vitro Biotransformation Studies}, series = {Cells}, volume = {12}, journal = {Cells}, number = {17}, issn = {2073-4409}, doi = {10.3390/cells12172140}, abstract = {In the liver, phase-1 biotransformation of drugs and other xenobiotics is largely facilitated by enzyme complexes consisting of cytochrome P450 oxidoreductase (CPR) and cytochrome P450 monooxygenases (CYPs). Generated from human liver-derived cell lines, recombinant in vitro cell systems with overexpression of defined phase-1 enzymes are widely used for pharmacological and toxicological drug assessment and laboratory-scale production of drug-specific reference metabolites. Most, if not all, of these cell lines, however, display some background activity of several CYPs, making it difficult to attribute effects to defined CYPs. The aim of this study was to generate cell lines with stable overexpression of human phase-1 enzymes based on Chinese hamster ovary (CHO) suspension cells. Cells were sequentially modified with cDNAs for human CPR in combination with CYP1A2, CYP2B6, or CYP3A4, using lentiviral gene transfer. In parallel, CYP-overexpressing cell lines without recombinant CPR were generated. Successful recombinant expression was demonstrated by mRNA and protein analyses. Using prototypical CYP-substrates, generated cell lines proved to display specific enzyme activities of each overexpressed CYP while we did not find any endogenous activity of those CYPs in parental CHO cells. Interestingly, cell lines revealed some evidence that the dependence of CYP activity on CPR could vary between CYPs. This needs to be confirmed in further studies. Recombinant expression of CPR was also shown to enhance CYP3A4-independent metabolisation of testosterone to androstenedione in CHO cells. We propose the novel serum-free CHO suspension cell lines with enhanced CPR and/or defined CYP activity as a promising "humanised" in vitro model to study the specific effects of those human CYPs. This could be relevant for toxicology and/or pharmacology studies in the pharmaceutical industry or medicine.}, language = {en} } @misc{RoggenbuckGoihlHanacketal., author = {Roggenbuck, Dirk and Goihl, Alexander and Hanack, Katja and Holzl{\"o}hner, Pamela and Hentschel, Christian and Veiczi, Miklos and Schierack, Peter and Reinhold, Dirk and Schulz, Hans-Ulrich}, title = {Serological diagnosis and prognosis of severe acute pancreatitis by analysis of serum glycoprotein 2}, series = {Clinical Chemistry and Laboratory Medicine}, volume = {55}, journal = {Clinical Chemistry and Laboratory Medicine}, number = {6}, issn = {1437-4331}, doi = {10.1515/cclm-2016-0797}, pages = {854 -- 864}, language = {en} } @misc{JurischkaDinterEfimovaetal., author = {Jurischka, Christoph and Dinter, Franziska and Efimova, Anastasia and Weiss, Romano and Schiebel, Juliane and Schulz, Christian and Fayziev, Bekzodjon and Schierack, Peter and Fischer, Thomas and R{\"o}diger, Stefan}, title = {An explorative study of polymers for 3D printing of bioanalytical test systems}, series = {Clinical Hemorheology and Microcirculation}, volume = {75}, journal = {Clinical Hemorheology and Microcirculation}, number = {1}, issn = {1875-8622}, doi = {10.3233/CH-190713}, pages = {57 -- 84}, abstract = {Background: The 3D printing is relevant as a manufacturing technology of functional models for forensic, pharmaceutical and bioanalytical applications such as drug delivery systems, sample preparation and point-of-care tests. Objective: Melting behavior and autofluorescence of materials are decisive for optimal printing and applicability of the product which are influenced by varying unknown additives. Methods: We have produced devices for bioanalytical applications from commercially available thermoplastic polymers using a melt-layer process. We characterized them by differential scanning calorimetry, fluorescence spectroscopy and functional assays (DNA capture assay, model for cell adhesion, bacterial adhesion and biofilm formation test). Results: From 14 tested colored, transparent and black materials we found only deep black acrylonitrile-butadiene-styrene (ABS) and some black polylactic acid (PLA) useable for fluorescence-based assays, with low autofluorescence only in the short-wave range of 300-400 nm. PLA was suitable for standard bioanalytical purposes due to a glass transition temperature of approximately 60°C, resistance to common laboratory chemicals and easy print processing. For temperature-critical methods, such as hybridization reactions up to 90°C, ABS was better suited. Conclusions: Autofluorescence was not a disadvantage per se but can also be used as a reference signal in assays. The rapid development of individual protocols for sample processing and analysis required the availability of a material with consistent quality over time. For fluorescence-based assays, the use of commercial standard materials did not seem to meet this requirement.}, language = {en} } @incollection{ElmerHohbergRusselletal., author = {Elmer, Michael and Hohberg, Karin and Russell, D. and Christian, Axel and Schulz, H.- J. and Wanner, Manfred}, title = {Succession of the soil faunal community during initial ecosystem development}, series = {Initial development of the artificial catchment "Chicken Creek" : monitoring program and survey 2005-2008}, booktitle = {Initial development of the artificial catchment "Chicken Creek" : monitoring program and survey 2005-2008}, editor = {Schaaf, Wolfgang and Biemelt, Detlef and H{\"u}ttl, Reinhard F.}, publisher = {Univ. of Technology, Research Center Landscape Development and Mining Landscapes}, address = {Cottbus}, pages = {97 -- 118}, language = {en} } @misc{MahmoodScharobaSchorlemeretal., author = {Mahmood, Safdar and Scharoba, Stefan and Schorlemer, Jonas and Schulz, Christian and H{\"u}bner, Michael and Reichenbach, Marc}, title = {Detecting Improvised Land-mines using Deep Neural Networks on GPR Image Dataset targeting FPGAs }, series = {IEEE Nordic Circuits and Systems Conference (NORCAS), 25-26 October 2022, Oslo, Norway}, journal = {IEEE Nordic Circuits and Systems Conference (NORCAS), 25-26 October 2022, Oslo, Norway}, publisher = {IEEE}, address = {Piscataway, NJ}, isbn = {979-8-3503-4550-6}, doi = {10.1109/norcas57515.2022.9934735}, pages = {1 -- 7}, language = {en} } @misc{KnauerSchulzZemellaetal., author = {Knauer, Jan Felix and Schulz, Christian and Zemella, Anne and W{\"u}stenhagen, Doreen A. and Walter, Ruben Magnus and K{\"u}pper, Jan-Heiner and Kubick, Stefan}, title = {Synthesis of mono Cytochrome P450 in a modified CHO-CPR cell-free protein production platform}, series = {Scientific Reports}, volume = {14}, journal = {Scientific Reports}, issn = {2045-2322}, doi = {10.1038/s41598-024-51781-6}, pages = {14}, abstract = {AbstractCytochromes P450 (CYPs) are a group of monooxygenases that can be found in almost all kinds of organisms. For CYPs to receive electrons from co-substrate NADPH, the activity of NADPH-Cytochrome-P450-oxidoreductase (CPR) is required as well. In humans, CYPs are an integral part of liver-based phase-1 biotransformation, which is essential for the metabolization of multiple xenobiotics and drugs. Consequently, CYPs are important players during drug development and therefore these enzymes are implemented in diverse screening applications. For these applications it is usually advantageous to use mono CYP microsomes containing only the CYP of interest. The generation of mono-CYP containing mammalian cells and vesicles is difficult since endogenous CYPs are present in many cell types that contain the necessary co-factors. By obtaining translationally active lysates from a modified CHO-CPR cell line, it is now possible to generate mono CYPs in a cell-free protein synthesis process in a straightforward manner. As a proof of principle, the synthesis of active human CYPs from three different CYP450 gene families (CYP1A2, CYP2B6 and CYP3A4), which are of outstanding interest in industry and academia was demonstrated. Luciferase based activity assays confirm the activity of the produced CYPs and enable the individual adaptation of the synthesis process for efficient cell-free enzyme production. Furthermore, they allow for substrate and inhibitor screenings not only for wild-type CYPs but also for mutants and further CYP isoforms and variants. As an example, the turnover of selected CYP substrates by cell-free synthesized CYPs was demonstrated via an indirect luciferase assay-based screening setup.}, language = {en} }