Publikationen
Hormonelle Aktivität in Wasser – Bestimmung hormoneller Aktivitäten in der aquatischen Umwelt mittels Arxula adeninivorans-Hefezellenassays
A. adeninivorans-Hefezellenassays sind nicht nur für die Beurteilung der Qualität eines Oberflächengewässers oder zur Einleiterkontrolle geeignet, sondern z.B. auch zur Überwachung einer Kläranlage und zur Beurteilung einer vierten Reinigungsstufe. Die unkomplizierten Assays sind besonders für kleine Labore interessant, die keine aufwändige Zellkultivierung leisten können. Mit transgenen Varianten der Hefe A. adeninivorans können hormonell aktive Substanzen bestimmt werden. (LABO 1-2/2018) Link zur Publikation
Effect-based trigger values for in vitro and in vivo bioassays performed on surface water extracts supporting the environmental quality standards (EQS) of the European Water Framework Directive
In ‘Science of the Total Environment 628–629 (2018) 748–765’ wird ein Verfahren zur Bestimmung der Effect-Based Trigger Values (EBT) für zahlreiche in vitro Bio-Assays vorgestellt. Auch für den A-YES® konnte mit dem beschriebenen Modell ein EBT für die Estrogenität berechnet werden. Effect-based trigger values for in vitro and in vivo bioassays performed on surface water extracts supporting the environmental quality standards (EQS) of the European Water Framework Directive Abstract Effect-based methods including cell-based bioassays, reporter gene assays and whole-organism assays have been applied for decades in water quality monitoring and testing of enriched solid-phase extracts. There is no common [...]
Validation of Arxula Yeast Estrogen Screen assay for detection of estrogenic activity in water samples: Results of an international interlaboratory study
In ‚Science of The Total Environment‘ werden in der April Ausgabe 2018 die Ergebnisse des Ringversuchs zur Validierung des Verfahrens A-YES® nach dem neuen Standard ISO19040-2 veröffentlicht. Abstract: Endocrine-active substances can adversely impact the aquatic ecosystems. A special emphasis is laid, among others, on the effects of estrogens and estrogen mimicking compounds. Effect-based screening methods like in vitro bioassays are suitable tools to detect and quantify endocrine activities of known and unknown mixtures. This study describes the validation of the Arxula-Yeast Estrogen Screen (A-YES®) assay, an effect-based method for the detection of the estrogenic potential of water and waste [...]
Separation and identification of hormone-active compounds using a combination of chromatographic separation and yeast-based reporter assay
Alexandre Chamas, Ha Thi Minh Pham, Martin Jähne, Karina Hettwer, Linda Gehrmann, JochenTuerk, Steffen Uhlig, Kirsten Simon, Kim Baronian, Gotthard Kunze Arxula adeninivorans-based yeast cell assays for the detection of steroid hormones demonstrated their efficiency for the determination of total hormone activity in a variety of samples using a microtiter plate format. In this study, a preliminary chromatographic separation using thin-layer chromatography plates is introduced in order to allow a rapid identification of the compounds responsible for this hormonal activity. The yeast whole cell assay can then be performed on the plate, producing a detectable signal where a [...]
Simultaneous detection of three sex steroid hormone classes using a novel yeast-based biosensor
Alexandre Chamas, Ha Thi Minh Pham, Martin Jähne, Karina Hettwer, Steffen Uhlig, Kirsten Simon, Almuth Einspanier, Kim Baronian, Gotthard Kunze A biosensor detecting estrogens, progestogens, and androgens in complex samples and in a single step is described. Three Arxula adeninivorans yeast strains were created, each strain producing a different recombinant human hormone receptor and a different fluorescent reporter protein. These strains were then mixed to create G1212/YRC102-hHR-fluo, the biological component of the biosensor. During incubation with G1212/YRC102-hHR-fluo, hormones present in a sample bind to their target receptor, which leads to the production of a specific fluorescent protein. Three [...]
LABO Fachartikel November 2016
Hier können Sie unseren kürzlich veröffentlichten Fachbeitrag zu Wasser-/Abwasseranalytik in der LABO lesen. Bei Fragen oder Interesse könne Sie sich jederzeit an uns wenden. Ihr new_diagnostics Team
The determination of pharmaceuticals in wastewater using a recombinant Arxula adeninivorans whole cell biosensor
Ha Thi Minh Pham, Martin Giersberg, Linda Gehrmann, Karina Hettwer, Jochen Tuerk, Steffen Uhlig, Gerold Hanke, Peter Weisswange, Kirsten Simon, Keith Baronian, Gotthard Kunze An Arxula adeninivorans based microbial biosensor has been developed for the determination of pharmaceuticals and chemicals such as omeprazole, lansoprazole, β-naphtoflavone and methylcholanthrene within 5 h using biochemical detection and 4 h and 10 min using amperometric detection. The biosensor consists of genetically modifiedA. adeninivorans G1212/YRC102-hAhR-hARNT-phyK (hAhR – human arylhydrocarbon receptor; hARNT – human arylhydrocarbon receptor nuclear translocator; inducible phyK– derived from Klebsiella sp. ASR1) as the biological component and coupled with either a biochemical or an amperometric detection method. The combination between hAhreceptor gene, hARNT and the [...]
Development of a recombinant Arxula adeninivorans cell bioassay for the detection of molecules with progesterone activity in wastewater
Chamas A, Nieter A, Pham HT, Giersberg M, Hettwer K, Uhlig S, Simon K, Baronian K, Kunze G This study describes the development of a bioassay to detect the presence of progesterone and progesterone-like molecules in wastewater samples. The basis of the bioassay is the integration of the human progesterone receptor gene into the yeast Arxula adeninivorans for the constitutive synthesis of the receptor. After incubation, binding of the analyte to the receptor induces the production of a reporter protein. Two reporter proteins were compared for detection parameters such as half-maximal activity (EC50), limit of detection (LoD) and limit [...]
Kinetic Analyses of Data from a Human Serum Albumin Assay Using the liSPR System
Anja Henseleit, Carolin Pohl, Hans-Michael Kaltenbach, Karina Hettwer, Kirsten Simon, Steffen Uhlig, Natalie Haustein, Thomas Bley and Elke Boschke We used the interaction between human serum albumin (HSA) and a high-affinity antibody to evaluate binding affinity measurements by the bench-top liSPR system (capitalis technology GmbH). HSA was immobilized directly onto a carboxylated sensor layer, and the mechanism of interaction between the antibody and HSA was investigated. The bivalence and heterogeneity of the antibody caused a complex binding mechanism. Three different interaction models (1:1 binding, heterogeneous analyte, bivalent analyte) were compared, and the bivalent analyte model best fit the curves [...]
Development and assessment of a novel Arxula adeninivorans androgen screen (A-YAS) assay and its application in analysis of cattle urine
Gerlach T, Knaust J, Kaiser C, Körner M, Hettwer K, Uhlig S, Simon K, Baronian K, Kunze G The novel A-YAS assay for the detection of androgenic activity in liquid samples such as urine has been developed and assessed. The assay is based on transgenic Arxula adeninivorans yeast cells as the bio-component. The cells were engineered to co-express the human androgen receptor (hAR) gene and the inducible phytase reporter gene (phyK, derived from Klebsiella sp. ASR1), under the control of an Arxula derived glucoamylase (GAA) promoter, which had been modified by the insertion of hormone-responsive elements (HREs). The Arxula transformation/ expression platformXplor®2 was used to select stablemitotic [...]
Projekte
Einführung eines QM-Systems nach ISO 9001
Wir haben es uns zur Aufgabe gemacht ein Qualitätsmanagementsystem einzuführen und zu etablieren, um uns für unsere Produkte und Dienstleistungen nach dem anerkannten Standard ISO 9001:2015 zu zertifizieren. Mit diesem Ziel erfolgt gleichzeitig die Vorbereitung für die Akkreditierung unseres Labors für den Einsatz unserer Bioassays als Prüfverfahren entsprechend den Anforderungen aus der ISO 17025:2018.
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