<?xml version="1.0" encoding="utf-8"?>
<export-example>
  <doc>
    <id>983</id>
    <completedYear>2017</completedYear>
    <publishedYear/>
    <thesisYearAccepted/>
    <language>eng</language>
    <pageFirst/>
    <pageLast/>
    <pageNumber/>
    <edition/>
    <issue>3</issue>
    <volume>4</volume>
    <type>article</type>
    <publisherName/>
    <publisherPlace/>
    <creatingCorporation/>
    <contributingCorporation/>
    <belongsToBibliography>1</belongsToBibliography>
    <completedDate>--</completedDate>
    <publishedDate>--</publishedDate>
    <thesisDateAccepted>--</thesisDateAccepted>
    <title language="eng">In Vitro Evaluation of Glycoengineered RSV-F in the Human Artificial Lymph Node Reactor</title>
    <abstract language="eng">Subunit vaccines often require adjuvants to elicit sustained immune activity. Here, a method is described to evaluate the efficacy of single vaccine candidates in the preclinical stage based on cytokine and gene expression analysis. As a model, the recombinant human respiratory syncytial virus (RSV) fusion protein (RSV-F) was produced in CHO cells. For comparison, wild-type and glycoengineered, afucosylated RSV-F were established. Both glycoprotein vaccines were tested in a commercial Human Artificial Lymph Node in vitro model (HuALN®). The analysis of six key cytokines in cell culture supernatants showed well-balanced immune responses for the afucosylated RSV-F, while immune response of wild-type RSV-F was more Th1 accentuated. In particular, stronger and specific secretion of interleukin-4 after each round of re-stimulation underlined higher potency and efficacy of the afucosylated vaccine candidate. Comprehensive gene expression analysis by nCounter gene expression assay confirmed the stronger onset of the immunologic reaction in stimulation experiments with the afucosylated vaccine in comparison to wild-type RSV-F and particularly revealed prominent activation of Th17 related genes, innate immunity, and comprehensive activation of humoral immunity. We, therefore, show that our method is suited to distinguish the potency of two vaccine candidates with minor structural differences.</abstract>
    <parentTitle language="eng">Bioengineering</parentTitle>
    <identifier type="issn">2306-5354</identifier>
    <identifier type="urn">urn:nbn:de:kobv:526-opus4-9831</identifier>
    <enrichment key="SourceTitle">Radke, L.; Sandig, G.; Lubitz, A.; Schließer, U.; von Horsten, H.H.; Blanchard, V.; Keil, K.; Sandig, V.; Giese, C.; Hummel, M.; Hinderlich, S.; Frohme, M.	In Vitro Evaluation of Glycoengineered RSV-F in the Human Artificial Lymph Node Reactor. Bioengineering 2017, 4, 70.</enrichment>
    <enrichment key="DOI_VoR">https://doi.org/10.3390/bioengineering4030070</enrichment>
    <licence>Creative Commons - CC BY - Namensnennung 4.0 International</licence>
    <author>Lars Radke</author>
    <author>Grit Sandig</author>
    <author>Annika Lubitz</author>
    <author>Ulrike Schließer</author>
    <author>Hans Henning von Horsten</author>
    <author>Véronique Blanchard</author>
    <author>Karolin Keil</author>
    <author>Volker Sandig</author>
    <author>Christoph Giese</author>
    <author>Michael Hummel</author>
    <author>Stephan Hinderlich</author>
    <author>Marcus Frohme</author>
    <subject>
      <language>eng</language>
      <type>uncontrolled</type>
      <value>glycoengineering</value>
    </subject>
    <subject>
      <language>eng</language>
      <type>uncontrolled</type>
      <value>fucosylation</value>
    </subject>
    <subject>
      <language>eng</language>
      <type>uncontrolled</type>
      <value>RSV</value>
    </subject>
    <subject>
      <language>eng</language>
      <type>uncontrolled</type>
      <value>F-protein</value>
    </subject>
    <subject>
      <language>eng</language>
      <type>uncontrolled</type>
      <value>NanoString</value>
    </subject>
    <collection role="ddc" number="570">Biowissenschaften; Biologie</collection>
    <collection role="institutes" number="">Fachbereich Ingenieur- und Naturwissenschaften</collection>
    <collection role="open_access" number="">open_access</collection>
    <collection role="green_open_access" number="1">Gold Open Access</collection>
    <thesisPublisher>Technische Hochschule Wildau</thesisPublisher>
    <file>https://opus4.kobv.de/opus4-th-wildau/files/983/bioengineering4030070.pdf</file>
  </doc>
  <doc>
    <id>962</id>
    <completedYear>2017</completedYear>
    <publishedYear/>
    <thesisYearAccepted/>
    <language>eng</language>
    <pageFirst/>
    <pageLast/>
    <pageNumber/>
    <edition/>
    <issue>2</issue>
    <volume>4</volume>
    <type>article</type>
    <publisherName/>
    <publisherPlace/>
    <creatingCorporation/>
    <contributingCorporation/>
    <belongsToBibliography>1</belongsToBibliography>
    <completedDate>--</completedDate>
    <publishedDate>--</publishedDate>
    <thesisDateAccepted>--</thesisDateAccepted>
    <title language="eng">Engineering of CHO Cells for the Production of Recombinant Glycoprotein Vaccines with Xylosylated N-glycans</title>
    <abstract language="eng">Xylose is a general component of O-glycans in mammals. Core-xylosylation of N-glycans is only found in plants and helminth. Consequently, xylosylated N-glycans cause immunological response in humans. We have used the F-protein of the human respiratory syncytial virus (RSV), one of the main causes of respiratory tract infection in infants and elderly, as a model protein for vaccination. The RSV-F protein was expressed in CHO-DG44 cells, which were further modified by co-expression of β1,2-xylosyltransferase from Nicotiana tabacum. Xylosylation of RSV-F N-glycans was shown by monosaccharide analysis and MALDI-TOF mass spectrometry. In immunogenic studies with a human artificial lymph node model, the engineered RSV-F protein revealed improved vaccination efficacy.</abstract>
    <parentTitle language="deu">Bioengineering</parentTitle>
    <identifier type="issn">2306-5354</identifier>
    <identifier type="urn">urn:nbn:de:kobv:526-opus4-9620</identifier>
    <enrichment key="SourceTitle">Sandig, G.; von Horsten, H.H.; Radke, L.; Blanchard, V.; Frohme, M.; Giese, C.; Sandig, V.; Hinderlich, S. Engineering of CHO Cells for the Production of Recombinant Glycoprotein Vaccines with Xylosylated N-glycans. Bioengineering 2017, 4, 38.</enrichment>
    <enrichment key="DOI_VoR">https://doi.org/10.3390/bioengineering4020038</enrichment>
    <licence>Creative Commons - CC BY - Namensnennung 4.0 International</licence>
    <author>Grit Sandig</author>
    <author>Hans Henning von Horsten</author>
    <author>Lars Radke</author>
    <author>Véronique Blanchard</author>
    <author>Marcus Frohme</author>
    <author>Christoph Giese</author>
    <author>Volker Sandig</author>
    <author>Stephan Hinderlich</author>
    <subject>
      <language>eng</language>
      <type>uncontrolled</type>
      <value>CHO</value>
    </subject>
    <subject>
      <language>eng</language>
      <type>uncontrolled</type>
      <value>glycoengineering</value>
    </subject>
    <subject>
      <language>eng</language>
      <type>uncontrolled</type>
      <value>respiratory syncytial virus</value>
    </subject>
    <subject>
      <language>eng</language>
      <type>uncontrolled</type>
      <value>vaccine</value>
    </subject>
    <subject>
      <language>eng</language>
      <type>uncontrolled</type>
      <value>xylose</value>
    </subject>
    <collection role="ddc" number="660">Chemische Verfahrenstechnik</collection>
    <collection role="institutes" number="">Fachbereich Ingenieur- und Naturwissenschaften</collection>
    <collection role="open_access" number="">open_access</collection>
    <collection role="green_open_access" number="1">Gold Open Access</collection>
    <thesisPublisher>Technische Hochschule Wildau</thesisPublisher>
    <file>https://opus4.kobv.de/opus4-th-wildau/files/962/bioengineering4020038.pdf</file>
  </doc>
  <doc>
    <id>115</id>
    <completedYear>2012</completedYear>
    <publishedYear>2012</publishedYear>
    <thesisYearAccepted/>
    <language>deu</language>
    <pageFirst>9</pageFirst>
    <pageLast>14</pageLast>
    <pageNumber/>
    <edition/>
    <issue/>
    <volume>16</volume>
    <type>articlewildau</type>
    <publisherName/>
    <publisherPlace/>
    <creatingCorporation/>
    <contributingCorporation/>
    <belongsToBibliography>1</belongsToBibliography>
    <completedDate>--</completedDate>
    <publishedDate>--</publishedDate>
    <thesisDateAccepted>--</thesisDateAccepted>
    <title language="deu">Einfluss der Kryokonservierung auf die Immunantwort von Leukozyten</title>
    <abstract language="deu">Die Messung von Zytokinen in Stimulationsexperimenten zur Bestimmung von Stärke und Umfang einer Immunreaktion werden in der Praxis häufig an kryokonserviertem Zellmaterial durchgeführt. Bisherige Untersuchungen zum Einfluss der Kryokonservierung auf die Zytokinexpression sind widersprüchlich. In den hier durchgeführten Experimenten wurden die Genexpression und/ oder Sekretion der Zytokine IL-2, IL-4, IL-5, IL-10, IL-12, IL-13, GM-CSF, IFN-γ und TNF-α in frisch isolierten und kryokonservierten Immunzellen aus Blut (PBMC) eines gesunden Spenders bestimmt. Diese wurden mit den Immunsystem-aktivierenden Stoffen OKT-3 und Concanavalin A (Con A) stimuliert und für 4 bzw. 24 h kultiviert. Ein Vergleich der frisch-isolierten und kryokonservierten PBMC in Stimulationsexperimenten zeigt, (1) dass die Messung der Genexpression genauere Einblicke zu Beginn der einsetzenden Immunreaktion verschafft, als die Messung der ausgeschütteten Zytokine, (2) dass durch Einfrieren die Immunreaktion insbesondere zu diesem Zeitpunkt beeinflusst wird und (3) dass zu späteren Zeitpunkten die Konzentrationsbestimmung der Zytokine im Zellkulturüberstand das Mittel der Wahl ist.</abstract>
    <abstract language="eng">The measurement of cytokines in stimulation experiments with the aim to determine the strength and the complexitiy of an immune reaction is commonly carried out on cryopreserved cells. Previous studies, investigating the impact of cryopreservation on the cytokine expression, are inconsistent in their findings. Experiments conducted in this study determine the gene expression and/or secretion of IL-2, IL-4, IL-5, IL-10, IL-12, IL-13, GM-CSF, IFN-γ and TNF-α in freshly isolated and cryopreserved PBMC of a healthy donor. The cells were stimulated with the substances OKT-3 and Concanavalin A (Con A) and cultured for 4h and 24h. The comparison of freshly isolated and cryopreserved cells in stimulation experiments showed (1) that the measurement of the gene expression offers a more accurate insight into the immune-reaction at early timepoints than the measurement of the secreted cytokines; (2) that freezing of the cells affect the immune response at this early stage and (3) that at later points the measurement of secreted proteins is the method of choice.</abstract>
    <parentTitle language="deu">Wissenschaftliche Beiträge 2012</parentTitle>
    <identifier type="urn">urn:nbn:de:kobv:526-opus-1363</identifier>
    <identifier type="opus3-id">136</identifier>
    <identifier type="doi">10.15771/0949-8214_2012_1_1</identifier>
    <identifier type="issn">0949-8214</identifier>
    <enrichment key="ZSTiteliD">16238</enrichment>
    <licence>Creative Commons - CC BY-NC-ND 3.0 DE - Namensnennung - Nicht-kommerziell - Keine Bearbeitung 3.0 Deutschland</licence>
    <author>Lars Radke</author>
    <author>Annika Lubitz</author>
    <author>Christoph Giese</author>
    <author>Marcus Frohme</author>
    <collection role="ddc" number="620">Ingenieurwissenschaften und zugeordnete Tätigkeiten</collection>
    <collection role="institutes" number="">Fachbereich Ingenieurwesen / Wirtschaftsingenieurwesen (bis 8/2014)</collection>
    <collection role="Publikationen_der_TH_Wildau" number="">Wissenschaftliche Beiträge</collection>
    <collection role="open_access" number="">open_access</collection>
    <thesisPublisher>Technische Hochschule Wildau</thesisPublisher>
    <file>https://opus4.kobv.de/opus4-th-wildau/files/115/WB2012_01_Frohme_Radke.pdf</file>
  </doc>
  <doc>
    <id>1416</id>
    <completedYear>2019</completedYear>
    <publishedYear/>
    <thesisYearAccepted/>
    <language>deu</language>
    <pageFirst/>
    <pageLast/>
    <pageNumber>86</pageNumber>
    <edition/>
    <issue/>
    <volume/>
    <type>doctoralthesis</type>
    <publisherName/>
    <publisherPlace/>
    <creatingCorporation/>
    <contributingCorporation/>
    <belongsToBibliography>1</belongsToBibliography>
    <completedDate>--</completedDate>
    <publishedDate>--</publishedDate>
    <thesisDateAccepted>2019-06-23</thesisDateAccepted>
    <title language="deu">Evaluierung der Potenz von glykosylierten Impfstoffen am Beispiel des RSV-F Proteins</title>
    <abstract language="deu">Trotz intensiver Forschung und immensen Bedarfs steht heutzutage kein RSV-Impfstoff zur Verfügung. Moderne Subunit-Vakzine bedürfen einer Adjuvantierung, um eine ausreichend starke Immunantwort zu generieren. Mit der Veränderung von Glykoslierungsmustern des RSV-F Proteins wird eine kovalent gekoppelte Adjuvantierung hergestellt, die über die Aktivierung des angeborenen Immunsystems eine umfassende Immunantwort generiert. In dieser Arbeit wurden rekombinant hergestellte Varianten des RSV-F Proteins mit defukosylierten oder xylosylierten N-Glykanen auf ihr Potential, das Immunsystem zu aktivieren, untersucht. Dazu wurden Zytokine aus Zellkulturüberständen quantifiziert und Methoden zur Genexpressionsanalyse stimulierter Zellen etabliert. In Vorbereitung auf Stimulationsexperimente mit PBMCs wurde der Einfluss der Kryokonservierung auf Zellen untersucht und die Zytokinexpression auf Gen- und Proteinebene korreliert. Im Rahmen der Stimulationsexperimente mit komplexen Zellgemischen (PBMC und mDC) wurden die stabil exprimierten Referenzgene SDHA, TBP und PPIA für die Etablierung einer validen Genexpressionsanalyse identifiziert. Dabei wurde eine mathematische Konvertierung quantitativer Transkriptzahlen aus dem Sonden-basierten direkten RNA-Nachweis (NanoString®) und der relativen Quantifizierung mittels qPCR entwickelt. Die Zytokinmuster in Zellkulturüberständen aus Langzeitstimulationsexperimenten, welche mit PBMC, mDC und den beiden Impfstoffvarianten in einem komplexen Perfusionsreaktorsystem (HuALN®) durchgeführt worden waren, wurden mittels automatisierter Bead-basierter Multiplexanalytik quantifiziert. Im Zusammenhang mit der Untersuchung umfangreicher Genpanels konnten verstärkte Immunreaktionen durch die glykosidisch veränderten RSV-F Proteine nachgewiesen werden. Die Defukosylierung führt zu einer breiten Aktivierung des zellulären und humoralen adaptiven Immunsystems. Das xylosylierte RSV-F aktiviert zusätzlich Komponenten des angeborenen Immunsystems. Zudem gibt es Hinweise darauf, dass durch den Grad der Defukosylierung das Verhältnis zwischen Th1 und Th2 basierter Antwort modifiziert werden kann. Mit den verwendeten und etablierten Methoden ist es möglich, die Potenz von Impfstoffkandidaten mit geringen strukturellen Veränderungen zu unterscheiden. Um die komplexen Stimulationsexperimente in einem höheren Durchsatz durchführen zu können, wurde an der Entwicklung eines miniaturisierten Bioreaktors mitgewirkt. Die Untersuchungen der Dichtheit des Reaktors mittels Fluoreszenzmikroskopie und der Biokompatibilität der einzelnen Komponenten hatten direkten Einfluss auf das Design des Reaktors, dessen Aufbau- und Verbindungstechnik sowie die Wahl der Materialien. Die Funktionalität konnte in verschiedenen 2D und 3D Kultivierungsansätzen über eine langanhaltende Zellviabilität bestätigt werden. Die verstärkte Modulierung der Immunantwort, hervorgerufen durch modifizierte Glykanstrukturen, zeugt von einer aussichtsreichen Methode, die auf andere Pathogene angewandt werden kann. Das höhere Durchsatzpotential des miniaturisierten Perfusionsreaktors (mehr Zeitpunkte, mehr Replikate) kann zukünftig einen präziseren Einblick in die immunologischen Vorgänge gewähren.</abstract>
    <abstract language="eng">Despite intense research and immense demand there is no RSV vaccine available today. Modern subunit vaccines require adjuvants to generate a sufficiently strong immune response. By editing the glycosylation pattern of the RSV-F Protein covalently coupled adjuvants are produced, which generate comprehensive immune responses via the activation of the innate immune system. In this work recombinantly produced variants of the RSV-F protein with defucosylated or xylosylated Nglycans were tested for their potential to activate the immune system. For that, cytokines from cell culture supernatants were quantified and methods for gene expression analysis of stimulated cells were established. In preliminary PBMC stimulation experiments the influence of cryopreservation on cells was examined and the expression of cytokines on gene and protein level was correlated. Within the scope of stimulation experiments with complex cell mixtures (PBMC and DC) the reference genes SDHA, TBP and PPIA were identified as stably expressed in order to establish a valid gene expression analysis. A mathematical conversion of quantitative transcript numbers from a probe-based direct RNA detection assay (NanoString®) and the relative quantification via qPCR was developed. The cytokine pattern in cell culture supernatants from long-term experiments performed with PBMC, mDC and each vaccine candidate in a highly complex perfusion reactor system (HuALN®) were quantified by automated bead-based multiplex analytics. In connection with the analysis of broad gene panels increased immune responses could be detected caused by the glycosidically altered RSV-F proteins. Defucosylation leads to a comprehensive activation of the immune system. Xylosylated RSV-F additionally activates components of the innate immune system. Furthermore, there is evidence that the ratio of the Th1 and Th2 based immune response can be modified by the grade of the defucosylation. The applied and established methods are suited to distinguish the potency of vaccine candidates with minor structural differences. In order to perform stimulation experiments in a higher throughput, this work contributed to the establishment of a miniaturized bioreactor. Leakage testing of the reactor via fluorescence microscopy and the examination of the biocompatibility of all single components directly influenced the reactor design, its assembly and connection techniques as well as the choice of materials. The reactors functionality was confirmed in different 2D and 3D cultivation approaches by means of a long-lasting high cell viability. The stronger modulated immune response evoked by modified glycosidic structures provides a promising method that can be applied to other pathogens, too. By its potential for higher throughput (more time points, more replicas) the miniaturized perfusion reactor could be able to provide a more precise insight into the immunological processes.</abstract>
    <identifier type="urn">urn:nbn:de:kobv:526-opus4-14162</identifier>
    <enrichment key="opus.source">publish</enrichment>
    <enrichment key="DOI_VoR">https://doi.org/10.17169/refubium-2606</enrichment>
    <enrichment key="SourceTitle">Radke, L. (2019). Evaluierung der Potenz von glykosylierten Impfstoffen am Beispiel des RSV-F Proteins. Berlin: Charité - Universitätsmedizin Berlin. doi:10.17169/refubium-2606</enrichment>
    <licence>Creative Commons - CC BY - Namensnennung 4.0 International</licence>
    <author>Lars Radke</author>
    <subject>
      <language>eng</language>
      <type>uncontrolled</type>
      <value>RSV-F</value>
    </subject>
    <subject>
      <language>eng</language>
      <type>uncontrolled</type>
      <value>Glycosylation</value>
    </subject>
    <subject>
      <language>eng</language>
      <type>uncontrolled</type>
      <value>vaccine</value>
    </subject>
    <subject>
      <language>eng</language>
      <type>uncontrolled</type>
      <value>xylosylation</value>
    </subject>
    <subject>
      <language>eng</language>
      <type>uncontrolled</type>
      <value>defucosylation</value>
    </subject>
    <collection role="ddc" number="610">Medizin und Gesundheit</collection>
    <collection role="institutes" number="">Fachbereich Ingenieur- und Naturwissenschaften</collection>
    <collection role="open_access" number="">open_access</collection>
    <thesisPublisher>Technische Hochschule Wildau</thesisPublisher>
    <thesisGrantor>Charité - Universitätsmedizin Berlin</thesisGrantor>
    <file>https://opus4.kobv.de/opus4-th-wildau/files/1416/Dissertation_RADKE_LARS.pdf</file>
  </doc>
  <doc>
    <id>352</id>
    <completedYear>2015</completedYear>
    <publishedYear/>
    <thesisYearAccepted/>
    <language>deu</language>
    <pageFirst>15</pageFirst>
    <pageLast>19</pageLast>
    <pageNumber/>
    <edition/>
    <issue/>
    <volume>19</volume>
    <type>articlewildau</type>
    <publisherName/>
    <publisherPlace/>
    <creatingCorporation/>
    <contributingCorporation/>
    <belongsToBibliography>1</belongsToBibliography>
    <completedDate>--</completedDate>
    <publishedDate>2015-03-23</publishedDate>
    <thesisDateAccepted>--</thesisDateAccepted>
    <title language="deu">Automatisierung des Bio-Plex Pro Analyseverfahrens</title>
    <abstract language="deu">Für den simultanen Nachweis mehrerer Analyten innerhalb einer Probe ist die Bead-basierte Multiplexanalytik ein häufig verwendetes Verfahren und wird beispielsweise zur Quantifizierung von Proteinen genutzt. Die aufwendige Durchführung der Assays soll durch Automatisierung einerseits dem Anwender abgenommen werden, während andererseits die Genauigkeit und Reproduzierbarkeit der Analyse gesteigert wird. Die Automatisierung des Bio-Plex Pro™ Assays ist mit dem Pipettierroboter Tecan Freedom EVO 200 umgesetzt worden. Es ist ein Skript mit der Software Freedom EVOware® entwickelt worden, welches die Probenvorbereitung des Assays vollständig übernimmt. Für einen Vergleich der manuellen und automatisierten Methode sind die humanen Zytokine IL-2, IL-4, IL-10, GM-CSF, IFN-γ und TNF-α in einer achtstufigen Standardverdünnungsreihe und in unterschiedlich konzentrierten Proben gemessen worden. Die Berechnung der Streuungen (Standardabweichung und Variationskoeffizient) der einzelnen Standardverdünnungsreihen sowie der Vergleich von gemessenen und erwarteten Konzentrationen der automatisierten und manuellen Methode zeigen, dass die Automatisierung neben der zeitlichen Optimierung auch die Genauigkeit und Reproduzierbarkeit der Analyse verbessert.</abstract>
    <abstract language="eng">Bead-based multpiplex analysis is frequently used for the simultaneous detection of multiple analytes within a sample. Such assays are commonly used to quantify proteins. The automation of the process relieves the user from the complex assay conductance and on the other hand increases the accuracy and reproducibility of the analysis. The automation of the Bio-Plex Pro™ assay has been successfully implemented with the pipetting-robot Tecan Freedom EVO 200. A script has been developed, using the Freedom EVOware® which has the ability to perform the complete assay procedure. For a comparison between manual and automated methods, the human cytokines IL-2, IL-4, IL-10, GM-CSF, IFN-γ and TNF-α were analysed by an eight-point standard dilution series and with samples of different concentrations. The calculation of the variances (standard deviation and coefficient of variation) of the single standard dilution series as well as the comparison of the observed and expected concentrations of the manual or automated method show the higher precision and reproducibility of the automated process in addition to its time-saving nature.</abstract>
    <parentTitle language="deu">Wissenschaftliche Beiträge 2015</parentTitle>
    <identifier type="doi">10.15771/0949-8214_2015_1_2</identifier>
    <identifier type="urn">urn:nbn:de:kobv:526-opus4-3524</identifier>
    <identifier type="issn">0949-8214</identifier>
    <enrichment key="review.accepted_by">2</enrichment>
    <enrichment key="ZSTiteliD">16238</enrichment>
    <licence>Creative Commons - CC BY-NC-ND 3.0 DE - Namensnennung - Nicht-kommerziell - Keine Bearbeitung 3.0 Deutschland</licence>
    <author>Karolin Keil</author>
    <author>Lars Radke</author>
    <author>Ulrich M. Tillich</author>
    <author>Marcus Frohme</author>
    <collection role="ddc" number="572">Biochemie</collection>
    <collection role="ddc" number="629">Andere Fachrichtungen der Ingenieurwissenschaften</collection>
    <collection role="Publikationen_der_TH_Wildau" number="">Wissenschaftliche Beiträge</collection>
    <collection role="institutes" number="">Fachbereich Ingenieur- und Naturwissenschaften</collection>
    <collection role="open_access" number="">open_access</collection>
    <collection role="green_open_access" number="3">Diamond Open Access</collection>
    <thesisPublisher>Technische Hochschule Wildau</thesisPublisher>
    <file>https://opus4.kobv.de/opus4-th-wildau/files/352/02_Automatisierung-des-Bio-Plex-Pro.pdf</file>
  </doc>
  <doc>
    <id>60</id>
    <completedYear>2010</completedYear>
    <publishedYear>2010</publishedYear>
    <thesisYearAccepted/>
    <language>deu</language>
    <pageFirst>6</pageFirst>
    <pageLast>12</pageLast>
    <pageNumber/>
    <edition/>
    <issue/>
    <volume>14</volume>
    <type>articlewildau</type>
    <publisherName/>
    <publisherPlace/>
    <creatingCorporation/>
    <contributingCorporation/>
    <belongsToBibliography>1</belongsToBibliography>
    <completedDate>--</completedDate>
    <publishedDate>--</publishedDate>
    <thesisDateAccepted>--</thesisDateAccepted>
    <title language="deu">Etablierung verschiedener Bead-basierter Multiplexmethoden mit einem Suspensions Array-System für molekulardiagnostische Zwecke</title>
    <abstract language="deu">Die simultane Bestimmung mehrerer Analyten und die Erstellung komplexer Parameterprofile erlangt immer größere Bedeutung in der heutigen Labordiagnostik. Die Bead-basierte Multiplexanalytik bietet hier eine flexible, schnelle und einfache Methode zur Erstellung individueller Analysen. Aufgrund der Vielzahl von Anwendungsmöglichkeiten, die diese moderne Nachweismethode im Bereich molekularbiologischer Fragestellungen bietet, ist die Etablierung der Beadbasierten Multiplexanalytik im Labor für Molekularbiologie und funktionelle Genomik der Technischen Hochschule Wildau von großem Nutzen. Zur Einarbeitung in das Testsystem wurden die Konzentrationen der Zytokine IL-2, IL-4, IFN-γ und TNF-α in Zellkulturüberständen mit kommerziellen Fertigsystemen gemessen und mit mRNA-Expressionsraten der gleichen Proben verglichen. Des Weiteren wurde ein Testsystem zum Nachweis von humanen Antikörpern der Klassen IgG und IgM sowie deren antigen-spezifischer Anteil in Zellkulturüberständen entwickelt. Außerdem konnte durch die erfolgreiche Detektion von DNA-gekoppelten Beads mittels markierter Oligonukleotidsequenz die Kopplung und die Anwendbarkeit der Methode auf Bindungsexperimente mit Nukleotidsequenzen gezeigt werden.</abstract>
    <abstract language="eng">The simultaneous determination of multiple analytes and the generation of complex parameter profiles gains increasing importance in today’s laboratory diagnostics. The bead-based multiplex assay is offering a flexible, rapid and easy to handle method for the creation of individual analyses. Because of the multitude of applications this modern detection system offers, the establishment of the bead-based multiplex technique is of great benefit for the Laboratory for Molecular Biology and Functional Genomics of the Technical University of Applied Sciences Wildau. To familiarize with the testing system the concentrations of the cytokines IL-2, IL-4, IFN-γ and TNF-α in cell culture supernatant were analysed with commercially available assays and compared with mRNA expression ratios of the same samples. Furthermore a custom testing system was developed for the detection of human IgG and IgM antibodies and also antigen specific antibodies in cell culture supernatants. The assignability of the method to binding experiments with nucleotide sequences could be shown by the successful detection of DNA-modified beads by conjugated oligonucleotides.</abstract>
    <parentTitle language="deu">Wissenschaftliche Beiträge 2009/2010</parentTitle>
    <identifier type="issn">0949-8214</identifier>
    <identifier type="urn">urn:nbn:de:kobv:526-opus-802</identifier>
    <identifier type="opus3-id">80</identifier>
    <identifier type="doi">10.15771/0949-8214_2010_1_1</identifier>
    <enrichment key="ZSTiteliD">16238</enrichment>
    <licence>Creative Commons - CC BY-NC-ND 3.0 DE - Namensnennung - Nicht-kommerziell - Keine Bearbeitung 3.0 Deutschland</licence>
    <author>Lars Radke</author>
    <author>Diana López-Hemmerling</author>
    <author>Annika Lubitz</author>
    <author>Christoph Giese</author>
    <author>Franz-Xaver Wildenauer</author>
    <author>Marcus Frohme</author>
    <collection role="ddc" number="571">Physiologie und verwandte Themen</collection>
    <collection role="ddc" number="572">Biochemie</collection>
    <collection role="institutes" number="">Fachbereich Ingenieurwesen / Wirtschaftsingenieurwesen (bis 8/2014)</collection>
    <collection role="Publikationen_der_TH_Wildau" number="">Wissenschaftliche Beiträge</collection>
    <collection role="open_access" number="">open_access</collection>
    <thesisPublisher>Technische Hochschule Wildau</thesisPublisher>
    <file>https://opus4.kobv.de/opus4-th-wildau/files/60/TH_WB2009_2010_S006_012_Radke_Frohme.pdf</file>
  </doc>
  <doc>
    <id>530</id>
    <completedYear>2016</completedYear>
    <publishedYear>2016</publishedYear>
    <thesisYearAccepted/>
    <language>deu</language>
    <pageFirst>23</pageFirst>
    <pageLast>29</pageLast>
    <pageNumber/>
    <edition/>
    <issue/>
    <volume>20</volume>
    <type>articlewildau</type>
    <publisherName/>
    <publisherPlace/>
    <creatingCorporation/>
    <contributingCorporation/>
    <belongsToBibliography>1</belongsToBibliography>
    <completedDate>2016-03-21</completedDate>
    <publishedDate>--</publishedDate>
    <thesisDateAccepted>--</thesisDateAccepted>
    <title language="deu">ECS-Komplex – ein neuer Biomarker bei Wachstumshormonstörungen?</title>
    <abstract language="deu">Störungen im Metabolismus des Wachstumshormons können sich als Wachstumshormonmangel oder -überschuss (Akromegalie) äußern. In beiden Fällen handelt es sich um seltene Krankheiten, die aufwendig behandelt werden müssen, da jeder Patient anders auf seine jeweilige Medikation anspricht. Zur Bestimmung des individuellen Ansprechverhaltens auf die medikamentöse Behandlung wird Insulin-like Growth Factor 1 verwendet, das jedoch als unzuverlässig für die Therapiekontrolle gilt. Um die Diagnostik und Therapie von Wachstumshormonstörungen zu verbessern, wurde in dieser Pilotstudie der Nutzen des neuen potenziellen Biomarkers Elongin B/C-Cullin5-Socs-box Komplex (ECS-Komplex) überprüft. Die fünf Proteine des Komplexes üben zusammen einen negativen Feedback-Mechanismus auf den Wachstumshormonrezeptor aus und regulieren ihn in Abhängigkeit vom Wachstumshormonspiegel im Blut. Für diese Pilotstudie wurden vier Patienten mit Wachstumshormondefizienz und 15 Patienten mit Akromegalie rekrutiert. Die Messung von Unterschieden in der Expression der Gene auf RNA-Ebene in Blutproben der Patienten erlaubt erste Aussagen über deren Eignung als therapeutische Marker für diese Krankheiten.</abstract>
    <abstract language="eng">Growth hormone (GH) dysfunctions can occur as a GH-deficiency (GHD) or an overproduction of GH, leading to acromegaly. Both are rare diseases, which have to be treated for many years before the correct individual dosage is found and a mitigation of symptoms can be achieved. Current medical therapy is determined by the levels of the insulin-like growth factor-1, which is considered to be an unreliable theranostic tool. In order to improve the diagnosis and therapy of patients with GH-dysfunctions, we investigated the benefits of the novel potential biomarker Elongin B/C-Cullin5-Socs-box complex (ECS-complex). Together, these proteins regulate the growth hormone receptor levels according to the blood GH concentration through a negative feedback loop. For this study, we were able to recruit four patients with a GHD and 15 patients&#13;
with acromegaly.&#13;
&#13;
The detected differential expression of the ECS-complex in patients with growth hormone dysfunctions allows for first conclusions about the potential of those proteins as predictive biomarker molecules for individualized therapies.</abstract>
    <parentTitle language="deu">Wissenschaftliche Beiträge 2016</parentTitle>
    <identifier type="issn">0949-8214</identifier>
    <identifier type="doi">10.15771/0949-8214_2016_1_3</identifier>
    <identifier type="urn">urn:nbn:de:kobv:526-opus4-5308</identifier>
    <enrichment key="ZSTiteliD">16238</enrichment>
    <enrichment key="DataCiteUrl">https://commons.datacite.org/doi.org/10.15771/0949-8214_2016_1_3</enrichment>
    <licence>Creative Commons - CC BY-NC-ND 3.0 DE - Namensnennung - Nicht-kommerziell - Keine Bearbeitung 3.0 Deutschland</licence>
    <author>Wilhelm Gossing</author>
    <author>Lars Radke</author>
    <author>Marcus Frohme</author>
    <author>Henrik Biering</author>
    <collection role="ddc" number="612">Humanphysiologie</collection>
    <collection role="ddc" number="616">Krankheiten</collection>
    <collection role="Publikationen_der_TH_Wildau" number="">Wissenschaftliche Beiträge</collection>
    <collection role="institutes" number="">Fachbereich Ingenieur- und Naturwissenschaften</collection>
    <collection role="open_access" number="">open_access</collection>
    <collection role="green_open_access" number="3">Diamond Open Access</collection>
    <thesisPublisher>Technische Hochschule Wildau</thesisPublisher>
    <file>https://opus4.kobv.de/opus4-th-wildau/files/530/ECS-Komplex.pdf</file>
  </doc>
  <doc>
    <id>1330</id>
    <completedYear>2020</completedYear>
    <publishedYear/>
    <thesisYearAccepted/>
    <language>eng</language>
    <pageFirst/>
    <pageLast/>
    <pageNumber/>
    <edition/>
    <issue>6</issue>
    <volume>8</volume>
    <type>article</type>
    <publisherName/>
    <publisherPlace/>
    <creatingCorporation/>
    <contributingCorporation/>
    <belongsToBibliography>1</belongsToBibliography>
    <completedDate>--</completedDate>
    <publishedDate>--</publishedDate>
    <thesisDateAccepted>--</thesisDateAccepted>
    <title language="eng">Biomarkers for Liquid Biopsies of Pituitary Neuroendocrine Tumors</title>
    <abstract language="eng">Pituitary neuroendocrine tumors (PitNET) do not only belong to the most common intracranial neoplasms but seem to be generally more common than has been thought. Minimally invasive liquid biopsies have the potential to improve their early screening efficiency as well as monitor prognosis by facilitating the diagnostic procedures. This review aims to assess the potential of using liquid biopsies of different kinds of biomarker species that have only been obtained from solid pituitary tissues so far. Numerous molecules have been associated with the development of a PitNET, suggesting that it often develops from the cumulative effects of many smaller genetic or epigenetic changes. These minor changes eventually pile up to switch critical molecules into tumor-promoting states, which may be the key regulatory nodes representing the most potent marker substances for a diagnostic test. Drugs targeting these nodes may be superior for the therapeutic outcome and therefore the identification of such pituitary-specific cellular key nodes will help to accelerate their application in medicine. The ongoing genetic degeneration in pituitary adenomas suggests that repeated tumor profiling via liquid biopsies will be necessary for personalized and effective treatment solutions.</abstract>
    <parentTitle language="eng">Biomedicines</parentTitle>
    <identifier type="issn">2227-9059</identifier>
    <identifier type="urn">urn:nbn:de:kobv:526-opus4-13307</identifier>
    <enrichment key="SourceTitle">Gossing, W.; Frohme, M.; Radke, L. Biomarkers for Liquid Biopsies of Pituitary Neuroendocrine Tumors. Biomedicines 2020, 8, 148.</enrichment>
    <enrichment key="DOI_VoR">https://doi.org/10.3390/biomedicines8060148</enrichment>
    <licence>Creative Commons - CC BY - Namensnennung 4.0 International</licence>
    <author>Wilhelm Gossing</author>
    <author>Marcus Frohme</author>
    <author>Lars Radke</author>
    <subject>
      <language>eng</language>
      <type>uncontrolled</type>
      <value>pituitary</value>
    </subject>
    <subject>
      <language>eng</language>
      <type>uncontrolled</type>
      <value>biomarker</value>
    </subject>
    <subject>
      <language>eng</language>
      <type>uncontrolled</type>
      <value>liquid biopsy</value>
    </subject>
    <subject>
      <language>eng</language>
      <type>uncontrolled</type>
      <value>cfRNA</value>
    </subject>
    <subject>
      <language>eng</language>
      <type>uncontrolled</type>
      <value>ctDNA</value>
    </subject>
    <subject>
      <language>eng</language>
      <type>uncontrolled</type>
      <value>miRNA</value>
    </subject>
    <subject>
      <language>eng</language>
      <type>uncontrolled</type>
      <value>epigenetics</value>
    </subject>
    <subject>
      <language>eng</language>
      <type>uncontrolled</type>
      <value>lncRNA</value>
    </subject>
    <collection role="ddc" number="570">Biowissenschaften; Biologie</collection>
    <collection role="institutes" number="">Fachbereich Ingenieur- und Naturwissenschaften</collection>
    <collection role="open_access" number="">open_access</collection>
    <collection role="Funding" number="">DFG-geförderter Publikationsfonds</collection>
    <collection role="green_open_access" number="1">Gold Open Access</collection>
    <thesisPublisher>Technische Hochschule Wildau</thesisPublisher>
    <file>https://opus4.kobv.de/opus4-th-wildau/files/1330/biomedicines-08-00148.pdf</file>
  </doc>
  <doc>
    <id>1380</id>
    <completedYear>2021</completedYear>
    <publishedYear/>
    <thesisYearAccepted/>
    <language>eng</language>
    <pageFirst/>
    <pageLast/>
    <pageNumber/>
    <edition/>
    <issue>2</issue>
    <volume>9</volume>
    <type>article</type>
    <publisherName>MDPI</publisherName>
    <publisherPlace/>
    <creatingCorporation/>
    <contributingCorporation/>
    <belongsToBibliography>1</belongsToBibliography>
    <completedDate>--</completedDate>
    <publishedDate>2021-02-17</publishedDate>
    <thesisDateAccepted>--</thesisDateAccepted>
    <title language="eng">The ElonginB/C-Cullin5-SOCS-Box-Complex Is a Potential Biomarker for Growth Hormone Disorders</title>
    <abstract language="eng">Insulin-like growth factor 1 (IGF-1) is the standard biochemical marker for the diagnosis and treatment control of acromegaly and growth hormone deficiency (GHD). However, its limitations necessitate the screening for new specific and sensitive biomarkers. The elonginB/C-cullin5-SOCS-box-complex (ECS-complex) (an intracellular five-protein complex) is stimulated by circulating growth hormone (GH) and regulates GH receptor levels through a negative feedback loop. It mediates the cells’ sensitivity for GH and therefore, represents a potent new biomarker for those diseases. In this study, individual ECS-complex proteins were measured in whole blood samples of patients with acromegaly (n = 32) or GHD (n = 12) via ELISA and compared to controls. Hierarchical clustering of the results revealed that by combining the three ECS-complex proteins suppressor of cytokine signaling 2 (SOCS2), cullin-5 and ring-box protein 2 (Rbx-2), 93% of patient samples could be separated from controls, despite many patients having a normal IGF-1 or not receiving medical treatment. SOCS2 showed the best individual diagnostic performance with an overall accuracy of 0.93, while the combination of the three proteins correctly identified all patients and controls. This resulted in perfect sensitivity and specificity for all patient groups, which demonstrates potential benefits of the ECS-complex proteins as clinical biomarkers for the diagnostics of GH-related diseases and substantiates their important role in GH metabolism.</abstract>
    <parentTitle language="eng">Biomedicines</parentTitle>
    <identifier type="issn">2227-9059</identifier>
    <identifier type="urn">urn:nbn:de:kobv:526-opus4-13802</identifier>
    <enrichment key="opus.import.date">2021-03-03T00:39:07+00:00</enrichment>
    <enrichment key="opus.source">sword</enrichment>
    <enrichment key="opus.import.user">sword</enrichment>
    <enrichment key="opus.import.file">attachment; filename=deposit.zip</enrichment>
    <enrichment key="opus.import.checksum">5f9612610681c311e8884bade974ab20</enrichment>
    <enrichment key="SourceTitle">Gossing, W.; Radke, L.; Biering, H.; Diederich, S.; Mai, K.; Frohme, M. The ElonginB/C-Cullin5-SOCS-Box-Complex Is a Potential Biomarker for Growth Hormone Disorders. Biomedicines 2021, 9, 201. https://doi.org/10.3390/biomedicines9020201</enrichment>
    <enrichment key="DOI_VoR">https://doi.org/10.3390/biomedicines9020201</enrichment>
    <licence>Creative Commons - CC BY - Namensnennung 4.0 International</licence>
    <author>Wilhelm Gossing</author>
    <author>Lars Radke</author>
    <author>Henrik Biering</author>
    <author>Sven Diederich</author>
    <author>Knut Mai</author>
    <author>Marcus Frohme</author>
    <subject>
      <language>eng</language>
      <type>uncontrolled</type>
      <value>acromegaly</value>
    </subject>
    <subject>
      <language>eng</language>
      <type>uncontrolled</type>
      <value>growth hormone deficiency</value>
    </subject>
    <subject>
      <language>eng</language>
      <type>uncontrolled</type>
      <value>biomarker</value>
    </subject>
    <subject>
      <language>eng</language>
      <type>uncontrolled</type>
      <value>SOCS2</value>
    </subject>
    <subject>
      <language>eng</language>
      <type>uncontrolled</type>
      <value>liquid biopsy</value>
    </subject>
    <subject>
      <language>eng</language>
      <type>uncontrolled</type>
      <value>pituitary</value>
    </subject>
    <collection role="ddc" number="570">Biowissenschaften; Biologie</collection>
    <collection role="institutes" number="">Fachbereich Ingenieur- und Naturwissenschaften</collection>
    <collection role="open_access" number="">open_access</collection>
    <collection role="Import" number="import">Import</collection>
    <collection role="Funding" number="">Publikationsfonds der TH Wildau</collection>
    <collection role="green_open_access" number="1">Gold Open Access</collection>
    <thesisPublisher>Technische Hochschule Wildau</thesisPublisher>
    <file>https://opus4.kobv.de/opus4-th-wildau/files/1380/biomedicines-09-00201-v2.pdf</file>
  </doc>
</export-example>
