TY - JOUR A1 - Eisold, Ursula A1 - Kupstat, Annette A1 - Klier, Dennis Tobias A1 - Primus, Philipp-A. A1 - Pschenitza, Michael A1 - Niessner, Reinhard A1 - Knopp, Dietmar A1 - Kumke, Michael Uwe T1 - Probing the physicochemical interactions of 3-hydroxy-benzo[a]pyrene with different monoclonal and recombinant antibodies by use of fluorescence line-narrowing spectroscopy JF - Analytical & bioanalytical chemistry N2 - Characterization of interactions between antigens and antibodies is of utmost importance both for fundamental understanding of the binding and for development of advanced clinical diagnostics. Here, fluorescence line-narrowing (FLN) spectroscopy was used to study physicochemical interactions between 3-hydroxybenzo[a]pyrene (3OH-BaP, as antigen) and a variety of solvent matrices (as model systems) or anti-polycyclic aromatic hydrocarbon antibodies (anti-PAH). We focused the studies on the specific physicochemical interactions between 3OH-BaP and different, previously obtained, monoclonal and recombinant anti-PAH antibodies. Control experiments performed with non-binding monoclonal antibodies and bovine serum albumin (BSA) indicated that nonspecific interactions did not affect the FLN spectrum of 3OH-BaP. The spectral positions and relative intensities of the bands in the FLN spectra are highly dependent on the molecular environment of the 3OH-BaP. The FLN bands correlate with different vibrational modes of 3OH-BaP which are affected by interactions with the molecular environment (pi-pi interactions, H-bonding, or van-der-Waals forces). Although the analyte (3OH-BaP) was the same for all the antibodies investigated, different binding interactions could be identified from the FLN spectra on the basis of structural flexibility and conformational multiplicity of the antibodies' paratopes. KW - FLNS KW - Antibody KW - Paratope KW - Hapten KW - Polycyclic aromatic hydrocarbons Y1 - 2014 U6 - https://doi.org/10.1007/s00216-013-7584-8 SN - 1618-2642 SN - 1618-2650 VL - 406 IS - 14 SP - 3387 EP - 3394 PB - Springer CY - Heidelberg ER - TY - JOUR A1 - Eisold, Ursula A1 - Sellrie, Frank A1 - Memczak, Henry A1 - Andersson, Anika A1 - Schenk, Jörg A. A1 - Kumke, Michael Uwe T1 - Dye tool box for a fluorescence enhancement immunoassay JF - Bioconjugate chemistry N2 - Immunochemical analytical methods are very successful in clinical diagnostics and are nowadays also emerging in the control of food as well as monitoring of environmental issues. Among the different immunoassays, luminescence based formats are characterized by their outstanding sensitivity making this format especially attractive for future applications. The need for multiparameter detection capabilities calls for a tool box of dye labels in order to transduce the biochemical reaction into an optically detectable signal. Here, in a multiparameter approach each analyte may be detected by a different dye with a unique emission color (covering the blue to red spectral range) or a unique luminescence decay kinetics. In the case of a competitive immunoassay format for each of the different dye labels an individual antibody would be needed. In the present paper a slightly modified approach is presented using a 7-aminocoumarin unit as the basic antigen against which highly specific antibodies were generated. Leaving the epitope region in the dyes unchanged but introducing a side group in positon 3 of the coumarin system allowed us to tune the optical properties of the coumarin dyes without the necessity of new antibody generation. Upon modification of the parent coumarin unit the full spectral range from blue to deep red was accessed. In the manuscript the photophysical characterization of the coumarin derivatives and their corresponding immunocomplexes with two highly specific antibodies is presented. The coumarin dyes and their immunocomplexes were characterized by steady-state and time-resolved absorption as well as emission spectroscopy. Moreover, fluorescence depolarization measurements were carried out to complement the data stressing the different binding modes of the two antibodies. The binding modes were evaluated using the photophysics of 7-aminocoumarins and how it was affected in the respective immunocomplexes, namely, the formation of the intramolecular charge transfer (ICT) as well as the twisted intramolecular charge transfer (TICT). In contrast to other antibody-dye pairs reported a distinct fluorescence enhancement upon formation of the antibody-dye complex up to a factor of SO was found. Because of the easy emission color tuning by tailoring the coumarin substitution for the antigen binding in nonrelevant position 3 of the parent molecule, a dye tool box is on hand which can be used in the construction of competitive multiparameter fluorescence enhancement immunoassays (FenIA). Y1 - 2018 U6 - https://doi.org/10.1021/acs.bioconjchem.7b00731 SN - 1043-1802 VL - 29 IS - 1 SP - 203 EP - 214 PB - American Chemical Society CY - Washington ER - TY - JOUR A1 - Eisold, Ursula A1 - Sellrie, Frank A1 - Schenk, Jörg A. A1 - Lenz, Christine A1 - Stöcklein, Walter F. M. A1 - Kumke, Michael Uwe T1 - Bright or dark immune complexes of anti-TAMRA antibodies for adapted fluorescence-based bioanalysis JF - Analytical & bioanalytical chemistry N2 - Fluorescence labels, for example fluorescein or rhodamin derivatives, are widely used in bioanalysis applications including lateral-flow assays, PCR, and fluorescence microscopy. Depending on the layout of the particular application, fluorescence quenching or enhancement may be desired as the detection principle. Especially for multiplexed applications or high-brightness requirements, a tunable fluorescence probe can be beneficial. The alterations in the photophysics of rhodamine derivatives upon binding to two different anti-TAMRA antibodies were investigated by absorption and fluorescence-spectroscopy techniques, especially determining the fluorescence decay time and steady-state and time-resolved fluorescence anisotropy. Two monoclonal anti-TAMRA antibodies were generated by the hybridoma technique. Although surface-plasmon-resonance measurements clearly proved the high affinity of both antibodies towards 5-TAMRA, the observed effects on the fluorescence of rhodamine derivatives were very different. Depending on the anti-TAMRA antibody either a strong fluorescence quenching (G71-DC7) or a distinct fluorescence enhancement (G71-BE11) upon formation of the immune complex was observed. Additional rhodamine derivatives were used to gain further information on the binding interaction. The data reveal that such haptens as 5-TAMRA could generate different paratopes with equal binding affinities but different binding interactions, which provide the opportunity to adapt bioanalysis methods including immunoassays for optimized detection principles for the same hapten depending on the specific requirements. KW - mAb KW - Fluorescence KW - Anisotropy KW - Exciplex KW - Energy-transfer probe Y1 - 2015 U6 - https://doi.org/10.1007/s00216-015-8538-0 SN - 1618-2642 SN - 1618-2650 VL - 407 IS - 12 SP - 3313 EP - 3323 PB - Springer CY - Heidelberg ER - TY - GEN A1 - Engelhard, Sonja A1 - Kumke, Michael Uwe A1 - Löhmannsröben, Hans-Gerd T1 - OPQS – optical process and quality sensing : exemplary applications in the beerbrewing and polyurethane foaming processes N2 - Optical methods play an important role in process analytical technologies (PAT). Four examples of optical process and quality sensing (OPQS) are presented, which are based on three important experimental techniques: near-infrared absorption, luminescence quenching, and a novel method, photon density wave (PDW) spectroscopy. These are used to evaluate four process and quality parameters related to beer brewing and polyurethane (PU) foaming processes: the ethanol content and the oxygen (O2) content in beer, the biomass in a bioreactor, and the cellular structures of PU foam produced in a pilot production plant. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - paper 004 KW - process analytical technology KW - beer KW - biomass KW - foam analysis KW - NIR spectroscopy KW - fluorescence quenching KW - photon density wave spectroscopy Y1 - 2006 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-12191 ER - TY - JOUR A1 - Engelhard, Sonja A1 - Kumke, Michael Uwe A1 - Löhmannsröben, Hans-Gerd T1 - Examples of the application of optical process and quality sensing (OPQS) to beer brewing and polyurethane foaming processes N2 - Optical methods play an important role in process analytical technologies (PAT). Four examples of optical process and quality sensing (OPQS) are presented, which are based on three important experimental techniques: near- infrared absorption, luminescence quenching, and a novel method, photon density wave (PDW) spectroscopy. These are used to evaluate four process and quality parameters related to beer brewing and polyurethane (PU) foaming processes: the ethanol content and the oxygen (O-2) content in beer, the biomass in a bioreactor, and the cellular structures of PU foam produced in a pilot production plant Y1 - 2006 UR - http://www.springerlink.com/content/100417 U6 - https://doi.org/10.1007/s00216-005-3364-4 SN - 1618-2642 ER - TY - GEN A1 - Frimmel, Fritz Hartmann A1 - Kumke, Michael Uwe T1 - Optische Parameter zur Stoffcharakterisierung vom Trinkwasser bis zum Abwasser N2 - Die Anwendung von optischen Parametern zur Stoffcharakterisierung wird diskutiert. Dabei ist der Schwerpunkt der Diskussion auf absorptions- und fluoreszenzspektroskopische Methoden gesetzt. Beide Methoden können schnell und zuverlässig – auch im on-line Betrieb – eingesetzt werden. Der Beitrag soll einen Überblick über die grundlegenden Möglichkeiten der Anwendung beider Methoden geben. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - paper 025 KW - Absorptionsspektroskopie KW - SAK KW - Fluoreszenzspektroskopie KW - Summenparameter KW - Huminstoffe KW - polyzyklische aromatische Kohlenwasserstoffe Y1 - 1998 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-13088 ER - TY - JOUR A1 - Gehne, Sören A1 - Flehr, Roman A1 - Kienzler, Andrea Altevogt Nee A1 - Berg, Maik A1 - Bannwarth, Willi A1 - Kumke, Michael Uwe T1 - Dye dynamics in three-color FRET samples JF - The journal of physical chemistry : B, Condensed matter, materials, surfaces, interfaces & biophysical chemistry N2 - Time-resolved emission data (fluorescence decay and fluorescence depolarization) of two three-color Forster resonance energy transfer (tc-FRET) systems consisting of a carbostyril donor (D), a ruthenium complex (Ru) as relay dye, and a Cy5 derivative (Cy) or, optionally, an anthraquinone quencher (Q) were carefully analyzed using advanced distribution analysis models. Thereby, it is possible to get information on the flexibility and mobility of the chromophores which are bound to double stranded (ds) DNA. Especially the distance distribution based on the analysis of the fluorescence depolarization is an attractive approach to complement data of fluorescence decay time analysis. The distance distributions extracted from the experimental data were in excellent agreement with those determined from accessible volume (AV) simulations. Moreover, the study showed that for tc-FRET systems the combination of dyes emitting on different time scales (e.g., nanoseconds vs microseconds) is highly beneficial in the distribution analysis of time-resolved luminescence data in cases where macromolecules such as DNA are involved. Here, the short lifetimes can yield information on the rotation of the dye molecule itself and the long lifetime can give insight in the overall dynamics of the macromolecule. Y1 - 2012 U6 - https://doi.org/10.1021/jp3064273 SN - 1520-6106 VL - 116 IS - 35 SP - 10798 EP - 10806 PB - American Chemical Society CY - Washington ER - TY - JOUR A1 - Gehne, Sören A1 - Sydow, Karl A1 - Dathe, Margitta A1 - Kumke, Michael Uwe T1 - Characterization of cell-penetrating lipopeptide micelles by spectroscopic methods JF - The journal of physical chemistry : B, Condensed matter, materials, surfaces, interfaces & biophysical chemistry N2 - The transport of bioactive compounds to the site of action is a great challenge. A promising approach to overcome application-related problems is the development of targeting colloidal transport systems, such as micelles which are equipped with uptake mediating moieties. Here, we investigated a set of novel lipopeptides which exhibit a surfactant-like structure due to attachment of two palmitoyl chains to the Nterminus of cationic or anionic amino acid sequences. We analyzed the association behavior of these lipopeptides by using 5(6)-carboxyfluorescein (CF)-labeled derivatives as a fluorescent probe and different spectroscopic methods such as fluorescence anisotropy and fluorescence correlation spectroscopy (FCS). The photophysical properties as well as the diffusion and rotational movements of the CF-labeled lipopeptides were exploited to determine the cmc and the size of the micelles consisting of lipopeptides. We could distinguish cationic and anionic lipopeptides by their association behavior and by studying the interactions with mouse brain capillary endothelial cells (b.end3). The cationic derivatives turned out to be very strong surfactants with a very low cmc in the micromolar range (0.5-14 mu M). The unique combination of micelle-forming property and cell-penetrating ability can pave the road for the development of a novel class of efficient drug carrier systems. Y1 - 2013 U6 - https://doi.org/10.1021/jp406053g SN - 1520-6106 VL - 117 IS - 46 SP - 14215 EP - 14225 PB - American Chemical Society CY - Washington ER - TY - JOUR A1 - Grunzel, Petra A1 - Pilarek, Maciej A1 - Steinbrueck, Doerte A1 - Neubauer, Antje A1 - Brand, Eva A1 - Kumke, Michael Uwe A1 - Neubauer, Peter A1 - Krause, Mirja T1 - Mini-scale cultivation method enables expeditious plasmid production in Escherichia coli JF - Biotechnology journal : systems & synthetic biology, nanobiotech, medicine N2 - The standard procedure in the lab for plasmid isolation usually involves a 2-mL, 16 h over-night cultivation in 15-mL bioreaction tubes in LB medium. This is time consuming, and not suitable for high-throughput applications. This study shows that it is possible to produce plasmid DNA (pDNA) in a 1.5-mL microcentrifuge tube with only 100 L cultivation volume in less than 7 h with a simple protocol. Compared with the standard LB cultivation for pDNA production reaching a final pDNA concentration range of 1.5-4 mu g mL(-1), a 6- to 10-fold increase in plasmid concentration (from 10 up to 25 mu g mL(-1) cultivation volume) is achieved using an optimized medium with an internal substrate delivery system (EnBase (R)). Different strains, plasmids, and the applicability of different inoculation tools (i.e. different starting ODs) were compared, demonstrating the robustness of the system. Additionally, dissolved oxygen was monitored in real time online, indicating that under optimized conditions oxygen limitation can be avoided. We developed a simple protocol with a significantly decreased procedure time, enabling simultaneous handling of more samples, while a consistent quality and a higher final pDNA concentration are ensured. KW - Escherichia coli KW - High-cell-density culture KW - Miniaturized cultivations KW - Optical oxygen sensor KW - Plasmid DNA production Y1 - 2014 U6 - https://doi.org/10.1002/biot.201300177 SN - 1860-6768 SN - 1860-7314 VL - 9 IS - 1 SP - 128 EP - 136 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Haubitz, Toni A1 - Drobot, Björn A1 - Tsushima, Satoru A1 - Steudtner, Robin A1 - Stumpf, Thorsten A1 - Kumke, Michael Uwe T1 - Quenching mechanism of uranyl(VI) by chloride and bromide in aqueous and non-aqueous solutions JF - The journal of physical chemistry : A, Molecules, spectroscopy, kinetics, environment & general theory N2 - A major hindrance in utilizing uranyl(VI) luminescence as a standard analytical tool, for example, in environmental monitoring or nuclear industries, is quenching by other ions such as halide ions, which are present in many relevant matrices of uranyl(VI) speciation. Here, we demonstrate through a combination of time-resolved laser-induced fluorescence spectroscopy, transient absorption spectroscopy, and quantum chemistry that coordinating solvent molecules play a crucial role in U(VI) halide luminescence quenching. We show that our previously suggested quenching mechanism based on an internal redox reaction of the 1:2-uranyl-halide-complex holds also true for bromide-induced quenching of uranyl(VI). By adopting specific organic solvents, we were able to suppress the separation of the oxidized halide ligand X-2(center dot-) and the formed uranyl(V) into fully solvated ions, thereby "reigniting" U(VI) luminescence. Time-dependent density functional theory calculations show that quenching occurs through the outer-sphere complex of U(VI) and halide in water, while the ligand-to-metal charge transfer is strongly reduced in acetonitrile. Y1 - 2021 U6 - https://doi.org/10.1021/acs.jpca.1c02487 SN - 1089-5639 SN - 1520-5215 VL - 125 IS - 20 SP - 4380 EP - 4389 PB - American Chemical Society CY - Washington ER -