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 - JOUR A1 - Schenk, Jörg A. A1 - Fettke, Jörg A1 - Lenz, Christine A1 - Albers, Katharina A1 - Mallwitz, Frank A1 - Gajovic-Eichelmann, Nenad A1 - Ehrentreich-Förster, Eva A1 - Kusch, Emely A1 - Sellrie, Frank T1 - Secretory leukocyte protease inhibitor (SLPI) might contaminate murine monoclonal antibodies after purification on protein G JF - Journal of biotechnology N2 - The large scale production of a monoclonal anti-progesterone antibody in serum free medium followed by affinity chromatography on protein G lead to a contamination of the antibody sample with a protein of about 14 kDa. This protein was identified by mass spectrometry as secretory leukocyte protease inhibitor (SLPI). This SLPI contamination lead to a failure of the fiber-optic based competitive fluorescence assay to detect progesterone in milk. Purification of the monoclonal antibody using protein A columns circumvented this problem. KW - Hybridoma KW - SLPI KW - Protein G KW - Progesterone KW - Serum-free Y1 - 2012 U6 - https://doi.org/10.1016/j.jbiotec.2011.12.025 SN - 0168-1656 VL - 158 IS - 1-2 SP - 34 EP - 35 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Dippong, Martin A1 - Carl, Peter A1 - Lenz, Christine A1 - Schenk, Jörg A. A1 - Hoffmann, Katrin A1 - Schwaar, Timm A1 - Schneider, Rudolf J. A1 - Kuhne, Maren T1 - Hapten-Specific Single-Cell Selection of Hybridoma Clones by Fluorescence-Activated Cell Sorting for the Generation of Monoclonal Antibodies JF - Analytical chemistry Y1 - 2017 U6 - https://doi.org/10.1021/acs.analchem.6b04569 SN - 0003-2700 SN - 1520-6882 VL - 89 SP - 4007 EP - 4012 PB - American Chemical Society CY - Washington ER -