UP Transfer GmbH

Potsdam, Germany

UP Transfer GmbH

Potsdam, Germany
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Sarauli D.,Wildau University of Applied Sciences | Peters K.,Ludwig Maximilians University of Munich | Xu C.,UP Transfer GmbH | Schulz B.,University of Potsdam | And 2 more authors.
ACS Applied Materials and Interfaces | Year: 2014

We report on the fabrication of a complex electrode architecture for efficient direct bioelectrocatalysis. In the developed procedure, the redox enzyme pyrroloquinoline quinone-dependent glucose dehydrogenase entrapped in a sulfonated polyaniline [poly(2-methoxyaniline-5-sulfonic acid)-co-aniline] was immobilized on macroporous indium tin oxide (macroITO) electrodes. The use of the 3D-conducting scaffold with a large surface area in combination with the conductive polymer enables immobilization of large amounts of enzyme and its efficient communication with the electrode, leading to enhanced direct bioelectrocatalysis. In the presence of glucose, the fabricated bioelectrodes show an exceptionally high direct bioelectrocatalytical response without any additional mediator. The catalytic current is increased more than 200-fold compared to planar ITO electrodes. Together with a high long-term stability (the current response is maintained for >90% of the initial value even after 2 weeks of storage), the transparent 3D macroITO structure with a conductive polymer represents a valuable basis for the construction of highly efficient bioelectronic units, which are useful as indicators for processes liberating glucose and allowing optical and electrochemical transduction. © 2014 American Chemical Society.

Sarauli D.,Wildau University of Applied Sciences | Xu C.,UP Transfer GmbH | Dietzel B.,A-D Technologies | Schulz B.,University of Potsdam | Lisdat F.,Wildau University of Applied Sciences
Journal of Materials Chemistry B | Year: 2014

A feasible approach to construct multilayer films of sulfonated polyanilines-PMSA1 and PABMSA1-containing different ratios of aniline, 2-methoxyaniline-5-sulfonic acid (MAS) and 3-aminobenzoic acid (AB), with the entrapped redox enzyme pyrroloquinoline quinone-dependent glucose dehydrogenase (PQQ-GDH) on Au and ITO electrode surfaces, is described. The formation of layers has been followed and confirmed by electrochemical impedance spectroscopy (EIS), which demonstrates that the multilayer assembly can be achieved in a progressive and uniform manner. The gold and ITO electrodes subsequently modified with PMSA1:PQQ-GDH and PABMSA1 films are studied by cyclic voltammetry (CV) and UV-Vis spectroscopy which show a significant direct bioelectrocatalytical response to the oxidation of the substrate glucose without any additional mediator. This response correlates linearly with the number of deposited layers. Furthermore, the constructed polymer/enzyme multilayer system exhibits a rather good long-term stability, since the catalytic current response is maintained for more than 60% of the initial value even after two weeks of storage. This verifies that a productive interaction of the enzyme embedded in the film of substituted polyaniline can be used as a basis for the construction of bioelectronic units, which are useful as indicators for processes liberating glucose and allowing optical and electrochemical transduction. © the Partner Organisations 2014.

Sarauli D.,Wildau University of Applied Sciences | Xu C.,UP Transfer GmbH | Dietzel B.,A-D Technologies | Schulz B.,University of Potsdam | Lisdat F.,Wildau University of Applied Sciences
Acta Biomaterialia | Year: 2013

Sulfonated polyanilines have become promising building blocks in the construction of biosensors, and therefore we use here differently substituted polymer forms to investigate the role of their structural composition and properties in achieving a direct electron transfer with the redox enzyme pyrroloquinoline quinone-dependent glucose dehydrogenase (PQQ-GDH). To this end, new copolymers containing different ratios of 2-methoxyaniline-5-sulfonic acid (MAS), 3-aminobenzenesulfonic acid (ABS) and 3-aminobenzoic acid (AB) units have been chemically synthesized. All polymers have been studied with respect to their ability to react directly with PQQ-GDH. This interaction has been monitored initially in solution, and subsequently on electrode surfaces. The results show that only copolymers with MAS and aniline units can directly react with PQQ-GDH in solution; the background can be mainly ascribed to the emeraldine salt redox state of the polymer, allowing rather easy reduction. However, when polymers and the enzyme are immobilized on the surface of carbon nanotube-containing electrodes, direct bioelectrocatalysis is also feasible in the case of copolymers composed of ABS/AB and MAS/AB units, existing initially in pernigraniline base form. This verifies that a productive interaction of the enzyme with differently substituted polymers is feasible when the electrode potential can be used to drive the reaction towards the oxidation of the substrate-reduced enzyme. These results clearly demonstrate that enzyme electrodes based on sulfonated polyanilines and direct bioelectrocatalysis can be successfully constructed. © 2013 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Sarauli D.,Wildau University of Applied Sciences | Tanne J.,Wildau University of Applied Sciences | Xu C.,UP Transfer GmbH | Schulz B.,University of Potsdam | And 2 more authors.
Physical Chemistry Chemical Physics | Year: 2010

The multilayer formation of two different sulfonated polyanilines with cytochrome c is presented and mechanistic aspects of the contributions of the polyelectrolytes' properties to the characteristics of the assemblies are discussed. These two modified polymers, PASA1 and PASA2 are chemically synthesized and differ in the grade of sulfonation, substitution, and the chain length of the polymer. The influence of these properties on the multilayer assembly with cytochrome c is studied in detail by Quartz Crystal Microbalance (QCM) technique and Cyclic Voltammetry (CV). It is shown that the multilayer formation is successful, however, the redox activity of polyanilines itself has to be taken into account. In the case of a strong redox activity (PASA2) voltammetric analysis allows the separation of redox processes addressed to the polyelectrolyte and cyt c. For multilayers with PASA1 as building block electroactivity can be predominantly attributed to cyt c ensuring a high amount of electroactive protein and a low probability for interfering redox reaction, making this system suitable for biosensor applications. © the Owner Societies 2010.

Inal S.,University of Potsdam | Kolsch J.D.,University of Potsdam | Sellrie F.,UP Transfer GmbH | Schenk J.A.,UP Transfer GmbH | And 4 more authors.
Journal of Materials Chemistry B | Year: 2013

We present two thermoresponsive water soluble copolymers prepared via free radical statistical copolymerization of N-isopropylacrylamide (NIPAm) and of oligo(ethylene glycol) methacrylates (OEGMAs), respectively, with a solvatochromic 7-(diethylamino)-3-carboxy-coumarin (DEAC)-functionalized monomer. In aqueous solutions, the NIPAm-based copolymer exhibits characteristic changes in its fluorescence profile in response to a change in solution temperature as well as to the presence of a specific protein, namely an anti-DEAC antibody. This polymer emits only weakly at low temperatures, but exhibits a marked fluorescence enhancement accompanied by a change in its emission colour when heated above its cloud point. Such drastic changes in the fluorescence and absorbance spectra are observed also upon injection of the anti-DEAC antibody, attributed to the specific binding of the antibody to DEAC moieties. Importantly, protein binding occurs exclusively when the polymer is in the well hydrated state below the cloud point, enabling a temperature control on the molecular recognition event. On the other hand, heating of the polymer-antibody complexes releases a fraction of the bound antibody. In the presence of the DEAC-functionalized monomer in this mixture, the released antibody competitively binds to the monomer and the antibody-free chains of the polymer undergo a more effective collapse and inter-aggregation. In contrast, the emission properties of the OEGMA-based analogous copolymer are rather insensitive to the thermally induced phase transition or to antibody binding. These opposite behaviours underline the need for a carefully tailored molecular design of responsive polymers aimed at specific applications, such as biosensing. © 2013 The Royal Society of Chemistry.

Stech M.,Fraunhofer Institute for Biomedical Engineering | Merk H.,RiNA GmbH | Schenk J.A.,UP Transfer GmbH | Schenk J.A.,Hybrotec GmbH | And 7 more authors.
Journal of Biotechnology | Year: 2013

Cell-free protein synthesis is of increasing interest for the rapid and high-throughput synthesis of many proteins, in particular also antibody fragments. In this study, we present a novel strategy for the production of single chain antibody fragments (scFv) in a eukaryotic in vitro translation system. This strategy comprises the cell-free expression, isolation and label-free interaction analysis of a model antibody fragment synthesized in two differently prepared insect cell lysates. These lysates contain translocationally active microsomal structures derived from the endoplasmic reticulum (ER), allowing for posttranslational modifications of cell-free synthesized proteins. Both types of these insect cell lysates enable the synthesis and translocation of scFv into ER-derived vesicles. However, only the one that has a specifically adapted redox potential yields functional active antibody fragments. We have developed a new methodology for the isolation of functional target proteins based on the translocation of cell-free produced scFv into microsomal structures and subsequent collection of protein-enriched vesicles. Antibody fragments that have been released from these vesicles are shown to be well suited for label-free binding studies. Altogether, these results show the potential of insect cell lysates for the production, purification and selection of antibody fragments in an easy-to-handle and time-saving manner. © 2012 Elsevier B.V.

Ahnert K.,University of Potsdam | Ahnert K.,Ambrosys GmbH | Abel M.,University of Potsdam | Abel M.,Ambrosys GmbH | And 3 more authors.
Journal of Materials Chemistry | Year: 2011

Wave energy harvesting could be a substantial renewable energy source without impact on the global climate and ecology, yet practical attempts have struggled with the problems of wear and catastrophic failure. An innovative technology for ocean wave energy harvesting was recently proposed, based on the use of soft capacitors. This study presents a realistic theoretical and numerical model for the quantitative characterization of this harvesting method. Parameter regions with optimal behavior are found, and novel material descriptors are determined, which dramatically simplify analysis. The characteristics of currently available materials are evaluated, and found to merit a very conservative estimate of 10 years for raw material cost recovery. This journal is © The Royal Society of Chemistry.

Kuhne M.,BAM Federal Institute of Materials Research and Testing | Dippong M.,BAM Federal Institute of Materials Research and Testing | Dippong M.,University of Potsdam | Flemig S.,BAM Federal Institute of Materials Research and Testing | And 6 more authors.
Journal of Immunological Methods | Year: 2014

A novel method that optimizes the screening for antibody-secreting hapten-specific hybridoma cells by using flow cytometry is described. Cell clones specific for five different haptens were analyzed. We selectively double stained and analyzed fixed hybridoma cells with fluorophore-labeled haptens to demonstrate the target-selectivity, and with a fluorophore-labeled anti-mouse IgG antibody to characterize the level of surface expression of membrane-bound IgGs. ELISA measurements with the supernatants of the individual hybridoma clones revealed that antibodies from those cells, which showed the highest fluorescence intensities in the flow cytometric analysis, also displayed the highest affinities for the target antigens. The fluorescence intensity of antibody-producing cells corresponded well with the produced antibodies' affinities toward their respective antigens. Immunohistochemical staining verified the successful double labeling of the cells. Our method makes it possible to perform a high-throughput screening for hybridoma cells, which have both an adequate IgG production rate and a high target affinity. © 2014 Elsevier B.V.

Sellrie F.,UP Transfer GmbH | Graser E.,AJ Innuscreen GmbH | Lenz C.,UP Transfer GmbH | Hillebrand T.,AJ Innuscreen GmbH | And 2 more authors.
Biosensors and Bioelectronics | Year: 2013

First homogenous immunoassay for sequence-specific nucleic acid detection is developed. The assay bases on our finding that a fluorophore inserted into a DNA probe instead of one of the internal nucleotides may get protected from fluorescence quenching caused by an anti-fluorophore antibody, if the probe is hybridized with the target sequence. This ensures a positive signal in the antibody presence. The assay enables quantitative detection and may have potential for development of homogenous high-throughput platforms. © 2012 Elsevier B.V.

Tan C.,Fraunhofer Institute for Cell Therapy and Immunology | Schenk J.A.,HybroTec GmbH | Schenk J.A.,UP Transfer GmbH | Gajovic-Eichelmann N.,Fraunhofer Institute for Cell Therapy and Immunology | And 2 more authors.
Talanta | Year: 2015

A new homogeneous immunoassay for the detection of progesterone was developed to measure its concentration in human serum. We utilized the weak cross-reactivity of a monoclonal anti-progesterone antibody to an analog molecule (in this case β-estradiol) to create a mixture, in which the fluorescence-labeled antibody (AbF) and quencher-labeled BSA-estradiol (eBSAq) were at optimized equilibrium. At this stage, most antibodies were bound to eBSAq and the fluorescence of AbF was quenched. After adding samples containing free progesterone to the system, these would replace the eBSAq at the antigen-binding site. The fluorescence would be released. In contrast to conventional competitive immunoassays, the fluorescence signal increases with increasing progesterone concentration, greatly simplifying detection and calibration. The performance of the assay was very simple; there was only one mixing step; and other hormones like testosterone, estradiol or dehydroepiandrosterone (DHEA) do not interfere the assay. A wide linear range from 0.1 μg/L to 100 μg/L was achieved in buffer, with a LOD of 0.1 μg/L. In human serum the LOD was 5 μg/L, and the linear range was 5-500 μg/L. For this assay it is important to find the right combination of antibody and cross-reactive antigen. If such a combination could be defined, it is conceivable to apply this assay to a wide range of analytes. © 2014 Elsevier B.V. All rights reserved.

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