TY - BOOK A1 - Wollenberger, Ursula A1 - Renneberg, Reinhard A1 - Bier, Frank Fabian A1 - Scheller, Frieder W. T1 - Analytische Biochemie : eine praktische Einführung in das Messen mit Biomolekülen Y1 - 2003 SN - 3-527-30166-6 PB - John Wiley & Sons CY - Hoboken ER - TY - JOUR A1 - Warmt, Christian A1 - Fenzel, Carolin Kornelia A1 - Henkel, Jörg A1 - Bier, Frank Fabian T1 - Using Cy5-dUTP labelling of RPA-amplicons with downstream microarray analysis for the detection of antibiotic resistance genes JF - Scientific reports N2 - In this report we describe Cy5-dUTP labelling of recombinase-polymerase-amplification (RPA) products directly during the amplification process for the first time. Nucleic acid amplification techniques, especially polymerase-chain-reaction as well as various isothermal amplification methods such as RPA, becomes a promising tool in the detection of pathogens and target specific genes. Actually, RPA even provides more advantages. This isothermal method got popular in point of care diagnostics because of its speed and sensitivity but requires pre-labelled primer or probes for a following detection of the amplicons. To overcome this disadvantages, we performed an labelling of RPA-amplicons with Cy5-dUTP without the need of pre-labelled primers. The amplification results of various multiple antibiotic resistance genes indicating great potential as a flexible and promising tool with high specific and sensitive detection capabilities of the target genes. After the determination of an appropriate rate of 1% Cy5-dUTP and 99% unlabelled dTTP we were able to detect the bla(CTX-M15) gene in less than 1.6E-03 ng genomic DNA corresponding to approximately 200 cfu of Escherichia coli cells in only 40 min amplification time. Y1 - 2021 U6 - https://doi.org/10.1038/s41598-021-99774-z SN - 2045-2322 VL - 11 IS - 1 PB - Macmillan Publishers Limited, part of Springer Nature CY - [London] ER - TY - JOUR A1 - Tanne, Johannes A1 - Jeoung, Jae-Hun A1 - Peng, Lei A1 - Yarman, Aysu A1 - Dietzel, Birgit A1 - Schulz, Burkhard A1 - Schad, Daniel A1 - Dobbek, Holger A1 - Wollenberger, Ursula A1 - Bier, Frank Fabian A1 - Scheller, Frieder W. T1 - Direct Electron Transfer and Bioelectrocatalysis by a Hexameric, Heme Protein at Nanostructured Electrodes JF - Electroanalysis : an international journal devoted to fundamental and practical aspects of electroanalysis N2 - A nanohybrid consisting of poly(3-aminobenzenesulfonic acid-co-aniline) and multiwalled carbon nanotubes [MWCNT-P(ABS-A)]) on a gold electrode was used to immobilize the hexameric tyrosine-coordinated heme protein (HTHP). The enzyme showed direct electron transfer between the heme group of the protein and the nanostructured surface. Desorption of the noncovalently bound heme from the protein could be excluded by control measurements with adsorbed hemin on aminohexanthiol-modified electrodes. The nanostructuring and the optimised charge characteristics resulted in a higher protein coverage as compared with MUA/MU modified electrodes. The adsorbed enzyme shows catalytic activity for the cathodic H2O2 reduction and oxidation of NADH. KW - HTHP KW - Nanohybrid KW - Poylaniline KW - Multiwalled carbon nanotube Y1 - 2015 U6 - https://doi.org/10.1002/elan.201500231 SN - 1040-0397 SN - 1521-4109 VL - 27 IS - 10 SP - 2262 EP - 2267 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Stöcklein, Walter F. M. A1 - Makower, Alexander A1 - Bier, Frank Fabian A1 - Scheller, Frieder W. T1 - Enzyme sensors and enzyme amplifification systems Y1 - 1997 ER - TY - JOUR A1 - Steffen, Jenny A1 - von Nickisch-Rosenegk, Markus A1 - Bier, Frank Fabian T1 - In vitro transcription of a whole gene on a surface-coupled template N2 - An artificial gene was constructed combining the T7 promoter and terminator with the EGFP-gene from the plasmid pEGFP. The functionality of the construct was shown by in vitro translation. The gene-construct was immobilised on a planar glass surface. The transcription was performed on the immobilised gene and mRNA was determined by RT-PCR. Multiple use of the immobilised gene was demonstrated Y1 - 2005 SN - 1473-0189 ER - TY - JOUR A1 - Stech, Marlitt A1 - Merk, Helmut A1 - Schenk, Jörg A. A1 - Stöcklein, Walter F. M. A1 - Wüstenhagen, Doreen Anja A1 - Micheel, Burkhard A1 - Duschl, Claus A1 - Bier, Frank Fabian A1 - Kubick, Stefan T1 - Production of functional antibody fragments in a vesicle-based eukaryotic cell-free translation system JF - Journal of biotechnology N2 - 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. KW - Cell-free KW - In vitro translation KW - Single chain antibody (scFv) KW - Insect lysate KW - Surface plasmon resonance Y1 - 2012 U6 - https://doi.org/10.1016/j.jbiotec.2012.08.020 SN - 0168-1656 VL - 164 IS - 2 SP - 220 EP - 231 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Stanke, Sandra A1 - Wenger, Christian A1 - Bier, Frank Fabian A1 - Hölzel, Ralph T1 - AC electrokinetic immobilization of influenza virus JF - Electrophoresis : microfluids & proteomics N2 - The use of alternating current (AC) electrokinetic forces, like dielectrophoresis and AC electroosmosis, as a simple and fast method to immobilize sub-micrometer objects onto nanoelectrode arrays is presented. Due to its medical relevance, the influenza virus is chosen as a model organism. One of the outstanding features is that the immobilization of viral material to the electrodes can be achieved permanently, allowing subsequent handling independently from the electrical setup. Thus, by using merely electric fields, we demonstrate that the need of prior chemical surface modification could become obsolete. The accumulation of viral material over time is observed by fluorescence microscopy. The influences of side effects like electrothermal fluid flow, causing a fluid motion above the electrodes and causing an intensity gradient within the electrode array, are discussed. Due to the improved resolution by combining fluorescence microscopy with deconvolution, it is shown that the viral material is mainly drawn to the electrode edge and to a lesser extent to the electrode surface. Finally, areas of application for this functionalization technique are presented. KW - AC electrokinetics KW - AC electroosmosis KW - dielectrophoresis KW - influenza virus KW - nanoelectrodes Y1 - 2022 U6 - https://doi.org/10.1002/elps.202100324 SN - 0173-0835 SN - 1522-2683 VL - 43 IS - 12 SP - 1309 EP - 1321 PB - Wiley-Blackwell CY - Weinheim ER - TY - JOUR A1 - Stanke, Sandra A1 - Bier, Frank Fabian A1 - Hoezel, Ralph T1 - Fluid streaming above interdigitated electrodes in dielectrophoresis experiments JF - Electrophoresis : microfluidics, nanoanalysis & proteomics N2 - For the investigation of alternating current electrokinetic effects, a system is presented that allows for the simultaneous observation of fluid flow above and around microelectrodes in all three directions in space. Beside the usual microscopical view from top, lateral observation through the same objective is made possible by two small mirrors that are placed next to the electrodes. Fluid flow and movement of fluorescent nanoparticles above interdigitated electrodes are monitored by fluorescence microscopy and digital imaging and are further analysed by image processing. Field frequencies are varied from 10 Hz to 1 GHz at up to 10V(rms). Electrical conductivity of the fluid is monitored in situ in the actual measuring chamber. KW - Dielectrophoresis KW - Fluid streaming KW - Nanobead Y1 - 2011 U6 - https://doi.org/10.1002/elps.201100096 SN - 0173-0835 VL - 32 IS - 18 SP - 2448 EP - 2455 PB - Wiley-Blackwell CY - Malden ER - TY - GEN A1 - Stanke, S. A1 - Wenger, C. A1 - Bier, Frank Fabian A1 - Hölzel, Ralph T1 - Dielectrophoretic functionalization of nanoelectrode arrays for the detection of influenza viruses T2 - European biophysics journal : with biophysics letters ; an international journal of biophysics Y1 - 2017 SN - 0175-7571 SN - 1432-1017 VL - 46 SP - S337 EP - S337 PB - Springer CY - New York ER - TY - JOUR A1 - Song, Min Ik A1 - Bier, Frank Fabian A1 - Scheller, Frieder W. T1 - A method to detect superoxide radicals using teflon membrane and superoxide dismutase Y1 - 1995 ER -