TY - GEN A1 - Tscheuschner, Georg A1 - Kaiser, Melanie N. A1 - Lisec, Jan A1 - Beslic, D. A1 - Muth, Thilo A1 - Krüger, M. A1 - Mages, H.-W. A1 - Dorner, B. G. A1 - Knospe, J. A1 - Schenk, J. A. A1 - Sellrie, F. A1 - Weller, Michael G. T1 - MALDI-TOF-MS-based identification of monoclonal murine anti-SARS-CoV-2 antibodies within one hour N2 - During the SARS-CoV-2 pandemic, many virus-binding monoclonal antibodies have been developed for clinical and diagnostic purposes. This underlines the importance of antibodies as universal bioanalytical reagents. However, little attention is given to the reproducibility crisis that scientific studies are still facing to date. In a recent study, not even half of all research antibodies mentioned in publications could be identified at all. This should spark more efforts in the search for practical solutions for the traceability of antibodies. For this purpose, we used thirty-five monoclonal antibodies against SARS-CoV-2 to demonstrate how sequence-independent antibody identification can be achieved by simple means applied onto the protein. First, we examined the intact and light chain masses of the antibodies relative to the reference material NIST-mAb 8671. Already half of the antibodies could be identified based solely on these two parameters. In addition, we developed two complementary peptide mass fingerprinting methods with MALDI-TOF-MS that can be performed in 45 minutes and had a combined sequence coverage of over 80%. One method is based on the partial acidic hydrolysis of the protein by 5 mM of sulfuric acid at 99 °C. Furthermore, we established a fast way for a tryptic digest without an alkylation step. We were able to show that the distinction of clones is possible simply by a brief visual comparison of the mass spectra. In this work, two clones originating from the same immunization gave the same fingerprints. Later, a hybridoma sequencing confirmed the sequence identity of these sister clones. In order to automate the spectral comparison for larger libraries of antibodies, we developed the online software ABID 2.0 (https://gets.shinyapps.io/ABID/). This open-source software determines the number of matching peptides in the fingerprint spectra. We propose that publications and other documents critically relying on monoclonal antibodies with unknown amino acid sequences should include at least one antibody fingerprint. By fingerprinting an antibody in question, its identity can be confirmed by comparison with a library spectrum at any time and context. KW - SARS-CoV-2 antibody KW - Reproducibility crisis KW - Peptide mass fingerprinting KW - Monoclonal antibody KW - Traceability KW - Antibody identification KW - Identity KW - Antibody light chain KW - MALDI-TOF-MS KW - Trypsin KW - Acidic cleavage KW - Antibody subclass KW - Database KW - Peak overlap KW - ABID KW - Sulfuric acid KW - Online software KW - Sequencing KW - Peptide coverage PY - 2022 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-545182 DO - https://doi.org/10.20944/preprints202203.0229.v1 SN - 2310-287X SP - 1 EP - 24 PB - MDPI CY - Basel AN - OPUS4-54518 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Tscheuschner, Georg A1 - Kaiser, Melanie N. A1 - Lisec, Jan A1 - Beslic, D. A1 - Muth, Thilo A1 - Krüger, M. A1 - Mages, H. W. A1 - Dorner, B. G. A1 - Knospe, J. A1 - Schenk, J. A. A1 - Sellrie, F. A1 - Weller, Michael G. T1 - MALDI-TOF-MS-Based Identification of Monoclonal Murine Anti-SARS-CoV-2 Antibodies within One Hour N2 - During the SARS-CoV-2 pandemic, many virus-binding monoclonal antibodies have been developed for clinical and diagnostic purposes. This underlines the importance of antibodies as universal bioanalytical reagents. However, little attention is given to the reproducibility crisis that scientific studies are still facing to date. In a recent study, not even half of all research antibodies mentioned in publications could be identified at all. This should spark more efforts in the search for practical solutions for the traceability of antibodies. For this purpose, we used 35 monoclonal antibodies against SARS-CoV-2 to demonstrate how sequence-independent antibody identification can be achieved by simple means applied to the protein. First, we examined the intact and light chain masses of the antibodies relative to the reference material NIST-mAb 8671. Already half of the antibodies could be identified based solely on these two parameters. In addition, we developed two complementary peptide mass fingerprinting methods with MALDI-TOF-MS that can be performed in 60 min and had a combined sequence coverage of over 80%. One method is based on the partial acidic hydrolysis of the protein by 5 mM of sulfuric acid at 99 degrees C. Furthermore, we established a fast way for a tryptic digest without an alkylation step. We were able to show that the distinction of clones is possible simply by a brief visual comparison of the mass spectra. In this work, two clones originating from the same immunization gave the same fingerprints. Later, a hybridoma sequencing confirmed the sequence identity of these sister clones. In order to automate the spectral comparison for larger libraries of antibodies, we developed the online software ABID 2.0. This open-source software determines the number of matching peptides in the fingerprint spectra. We propose that publications and other documents critically relying on monoclonal antibodies with unknown amino acid sequences should include at least one antibody fingerprint. By fingerprinting an antibody in question, its identity can be confirmed by comparison with a library spectrum at any time and context. KW - Reproducibility KW - Quality control KW - Traceability KW - Peptides KW - Peptide mass fingerprinting KW - Monoclonal antibody KW - Recombinant antibody KW - Identity KW - Antibody identification KW - Sequencing KW - Light chain KW - Mass spectrometry KW - Software KW - Open science KW - Library KW - COVID-19 KW - Corona virus KW - Sequence coverage KW - NIST-mAb 8671 KW - Reference material KW - RBD KW - Spike protein KW - Nucleocapsid KW - Cleavage KW - Tryptic digest KW - MALDI KW - DHAP KW - 2,5-dihydroxyacetophenone KW - Github KW - Zenodo KW - ABID PY - 2022 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-547347 DO - https://doi.org/10.3390/antib11020027 VL - 11 IS - 2 SP - 1 EP - 22 PB - MDPI CY - Basel AN - OPUS4-54734 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Neuhold, S. A1 - Algermissen, D. A1 - Drissen, P. A1 - Adamczyk, Burkart A1 - Presoly, P. A1 - Sedlazeck, K. P. A1 - Schenk, J. A1 - Raith, J. G. A1 - Pomberger, R. A1 - Vollprecht, D. T1 - Tailoring the FeO/SiO2 Ratio in Electric Arc Furnace Slags to Minimize the Leaching of Vanadium and Chromium N2 - Based on recently published research on leaching control mechanisms in electric arc furnace (EAF) slags, it is assumed that a FeO/SiO2 ratio of around one leads to low leached V and Cr concentrations. This ratio influences the mineral phase composition of the slag toward higher amounts of spinel and a lower solubility of calcium silicate phases by suppressing the formation of magnesiowuestite and highly soluble calcium silicate phases. To evaluate this hypothesis, laboratory and scaled up tests in an EAF pilot plant were performed on slag samples characterized by elevated V and Cr leaching and a high FeO/SiO2 ratio. Prior to the melting experiments, the optimum FeO/SiO2 ratio was calculated via FactSageTM. In the melting experiments, the ratio was adjusted by adding quartz sand, which also decreased the basicity (CaO/SiO2) of the slag. As a reference, remelting experiments without quartz sand addition were conducted and additionally, the influence of the cooling rate of the slag was examined. The remelted (without quartz sand) and the remelted modified slags (with quartz sand) were analyzed chemically and mineralogically and the leaching behavior was investigated. The modification of the slags yielded a minimized release of V and Cr, supporting the hypothesis that the FeO/SiO2 ratio influences the mineralogy and the leaching behavior. KW - Electric arc furnace (EAF) slags KW - FactSageTM calculations KW - Leaching control mechanisms KW - Melting experiments PY - 2020 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-571517 DO - https://doi.org/10.3390/app10072549 VL - 10 IS - 7 SP - 1 EP - 18 PB - MDPI AN - OPUS4-57151 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Erhard, Anton A1 - Wüstenberg, Hermann A1 - Schenk, G. A1 - Möhrle, W. A1 - Schulz, Eduard T1 - Messung der Risstiefe im Reflexions- und Durchschallungsverfahren mit fokussierten Schallfeldern T2 - DGZfP Vortragstagung CY - Saarbrücken, Germany DA - 1982-05-17 PY - 1982 AN - OPUS4-18814 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Erhard, Anton A1 - Schenk, Gottfried A1 - Gohlke, Dirk A1 - Engl, G. A1 - Fischer, E T1 - The SAPHIR and MINI-SAPHIR System - Versatile Tools for Advanced Solutions for the PWR Primary Circuit Inspection T2 - First International Conference on NDE in relation to Structural Integrity for Nuclear and Pressurised Components Amsterdam CY - Amsterdam, Netherlands DA - 1998-10-20 PY - 1998 AN - OPUS4-18875 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Wüstenberg, Hermann A1 - Schenk, G. A1 - Haufe, U. T1 - Dynamische Fokussierung in Schrägeinschallung bei der Ultraschallprüfung T2 - 4. Internationale Konferenz über Zerstörungsfreie Prüfung in der Kerntechnik CY - Lindau, Germany DA - 1981-05-25 PY - 1981 N1 - Serientitel: Berichtsband / Deutsche Gesellschaft für zerstörungsfreie Prüfung e. V. – Series title: Berichtsband / Deutsche Gesellschaft für zerstörungsfreie Prüfung e. V. IS - 8 SP - 1 EP - 15 PB - DGZfP CY - Berlin AN - OPUS4-33345 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Adamczyk, Burkart A1 - Neuhold, S. A1 - Drissen, P. A1 - Algermissen, D. A1 - Presoly, P. A1 - Sedlazeck, K. P. A1 - Schenk, J. A1 - Raith, J. G. A1 - Pomberger, R. A1 - Vollprecht, D. T1 - Tailoring the FeO/SiO2 ratio in electric arc furnace slags to minimize the leaching of Vanadium and Chromium N2 - Based on recently published research on leaching control mechanisms in electric arc furnace (EAF) slags, it is assumed that a FeO/SiO2 ratio of around one leads to low leached V and Cr concentrations. This ratio influences the mineral phase composition of the slag toward higher amounts of spinel and a lower solubility of calcium silicate phases by suppressing the formation of magnesiowuestite and highly soluble calcium silicate phases. To evaluate this hypothesis, laboratory and scaled up tests in an EAF pilot plant were performed on slag samples characterized by elevated V and Cr leaching and a high FeO/SiO2 ratio. Prior to the melting experiments, the optimum FeO/SiO2 ratio was calculated via FactSageTM. In the melting experiments, the ratio was adjusted by adding quartz sand, which also decreased the basicity (CaO/SiO2) of the slag. As a reference, remelting experiments without quartz sand addition were conducted and additionally, the influence of the cooling rate of the slag was examined. The remelted (without quartz sand) and the remelted modified slags (with quartz sand) were analyzed chemically and mineralogically and the leaching behavior was investigated. The modification of the slags yielded a minimized release of V and Cr, supporting the hypothesis that the FeO/SiO2 ratio influences the mineralogy and the leaching behavior. KW - Electric arc furnace slags KW - FactSage calculations KW - EAF slags KW - Leaching control mechanisms KW - Melting experiments PY - 2020 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-507748 DO - https://doi.org/10.3390/app10072549 VL - 10 IS - 7 SP - 2549 EP - 2565 AN - OPUS4-50774 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Carvalho, José Joao A1 - Walter, Astrid A1 - Baermann, Yvonne A1 - Weller, Michael G. A1 - Panne, Ulrich A1 - Schenk, J.A. A1 - Schneider, Rudolf T1 - Non-invasive monitoring of immunization progress in mice via IgG from feces N2 - A non-invasive method to monitor the humoral immune response in mice after immunization is described. From fecal pellets of an individual mouse, a sufficient amount of active immunoglobulins or their fragments can be extracted to perform a regular examination of the status of the immune response by immunoassay. Hapten-specific antibodies from the feces of mice from three immunization trials showed very similar characteristics to those obtained from serum at a given date. Therefore, it can be suspected that some serum IgG enters the intestinal lumen and ends up in the feces, where they appear to be considerably stable. Hapten-specific IgAs were not found in the feces. Being able to analyze antibody titers in feces could be an interesting animal welfare refinement to standard practice that does not entail repeated blood sampling. KW - Immunization monitoring KW - IgG KW - Coproantibodies KW - Feces KW - Animal welfare KW - 3R concept PY - 2012 UR - http://iv.iiarjournals.org/content/26/1/63.full.pdf+html SN - 0258-851x VL - 26 IS - 1 SP - 63 EP - 70 PB - In vivo CY - Athens AN - OPUS4-25283 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -