TY - CONF A1 - Schneider, Rudolf A1 - Konthur, Zoltán A1 - Jaeger, Carsten T1 - Reference Material (IgG) for CCQM-PAWG Key Comparison N2 - BAM proposes a recombinant human IgG (anti-tetanus monoclonal antibody SA13) as candidate material for the 2027 CCQM-PAWG key comparison on antibody purity. The antibody is produced via hybridoma and recombinant expression systems, with detailed sequence and glycosylation profiling. Analytical methods include MALDI-TOF-MS, SEC, DLS, and UV-vis spectroscopy to assess purity, aggregation, and stability. Preliminary results show high purity, low heterogeneity, and consistent size distribution. A minimum of 40 mg IgG is required, with 100 mg preferred for deeper characterization. Production is expected to be completed by the end of 2026, with potential for a joint technical project. T2 - Spring Meeting of CCQM-PAWG CY - Sèvres, France DA - 06.04.2025 KW - Antibodies KW - Research Grade Test Material KW - Mass spectrometry KW - Affinity PY - 2025 AN - OPUS4-63503 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schneider, Rudolf T1 - Monitoring trace substances in wastewater and surface water by immunoanalytical assays and sensors N2 - Monitoring the elimination of trace substances in wastewater treatment plants, as required in the recast of the EU Urban Wastewater Treatment Directive and the new Kommunalabwasserrichtlinie (KARL), creates a need for on-site analyses in wastewater treatment plants. Immunoanalytical methods can be carried out on site, e.g. in the form of rapid tests, but continuously operating (immuno-bio)sensors would be ideal. And antibody-based methods have the inherent disadvantage that one antibody must be used per analyte and therefore only a small number of substances can be analysed. A set of indicators has been integrated in the directives in accordance with the list of the Kompetenzzentrum Spurenstoffe BW (KomS) and offer points of reference here. Yet, in order to be able to use immunoanalytical methods for their monitoring, (DIN/ISO) standards are required that define the necessary quality requirements and quality management measures for the sensors. In addition, for the larger part of the indicators, the required antibodies have yet to be developed and made commercially available in the long term. The talk will highlight our previous work on high-throughput methods, such as ELISA, regarding the presence of anthropogenic markers and some indicators as well as research approaches for sensor technology that could be used on-site, and will provide an overview of deficits and future efforts. T2 - Kolloquium der Abteilung Technische Umweltchemie und Sensortechnik CY - Stuttgart, Germany DA - 21.05.2025 KW - Antibodies KW - Immunoassay KW - Microfluidics KW - Biosensor KW - Trace substances PY - 2025 AN - OPUS4-63501 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schneider, Rudolf T1 - Keeping the Y N2 - The talk traces my career path and presents BAM as an attractive employer. It also offers advice on shaping one's own career after PhD. T2 - IWC Alumni Day 2025 CY - Garching bei München, Germany DA - 04.07.2025 KW - Antibodies KW - ELISA KW - Career PY - 2025 AN - OPUS4-63640 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Vergin, Chantal C. T1 - Development of antibodies against mycotoxins and their application in immunochemical methods N2 - Food safety is a central and topical issue in our society and is governed by food law. Mycotoxins are secondary metabolic products formed by molds. These contaminants can enter food and thus the food chain through infestation, posing a serious health risk to humans and animals. For this reason, the European Commission has issued Regulation (EU) 2023/915, which sets maximum levels for certain mycotoxins in food. Currently, around 25 % of foodstuffs are contaminated with mycotoxins above the legally prescribed limits. Regular checks are essential to prevent such exceedances. The analytical methods currently available, mainly based on chromatographic techniques such as LC-MS/MS, are considered inadequate for on-site use – i.e., at processing and production facilities in the food industry – because they are technically complex and labor-intensive. One possible improvement is the use of immunoassays, which are widely accepted and employed in medical diagnostics. However, a basic prerequisite for developing such assays is the availability of specific antibodies. Our focus in this project is the development of high-affinity, highly selective monoclonal antibodies against mycotoxins for which either no antibodies, only polyclonal antibodies, or antibodies with insufficient specificity are currently available. The goal is to generate antibodies targeting patulin, Alternaria toxins, and ergot alkaloids for use in rapid tests and on-site analytical systems. A key aspect is the group selectivity of the antibodies with regard to the various ergot alkaloids. Depending on the mycotoxin, heterologous or homologous haptens are used to immunize mice. To ensure monoclonality, we apply limiting dilution and antigen-specific fluorescence-activated cell sorting (FACS) during the selection process. The antibodies developed will be used to expand SAFIA Technologies GmbH’s existing mycotoxin test-kit, enabling reliable detection of mycotoxins in food and thereby contributing to improved food safety standards. We will showcase our approach along with the results achieved so far T2 - 5th European Biosensor Symposium CY - Tarragona, Spain DA - 26.10.2025 KW - Antibodies KW - Biosensor KW - Immunoassay KW - Mycotoxins PY - 2025 AN - OPUS4-64459 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schneider, Rudolf T1 - Immunochemical sensing of key pollution indicators in the water cycle N2 - Organic micropollutants - often termed emerging contaminants - are increasingly detected throughout the water cycle, from drinking water to wastewater. Monitoring their distribution requires trace analytical methods that are fast, cost-effective, matrix-tolerant, and ideally portable. Immunoanalytical techniques, using antibodies, nanobodies, or fragments, offer high selectivity and sensitivity, though typically in single-analyte or oligoplexing formats. To maximize their utility, environmentally relevant indicator compounds should be selected to reflect pollutant input, partial removal during treatment, and persistence in wastewater and surface water. ELISA remains a robust method for high-throughput analysis. We have assays available for the com-pounds carbamazepine, diclofenac, cetirizine, estrone, sulfamethoxazole, caffeine, cocaine, bisphenol A, and isolithocholic acid. Simpler formats like fluorescence polarization immunoassays (FPIA) and lateral flow immunoassays (LFIA)—familiar from COVID-19 rapid tests — enable on-site screening. Multiplexing is feasible with bead-based microarrays and electrochemical sensors on microfluidic platforms, which additionally offer portability and the latter even independence from optical systems. Despite these advances, the development of highly selective, high-affinity binders remains a critical bottleneck. Promising strategies to overcome this include computational hapten design, nanobody engineering, and in silico optimization of binder sequences—enhanced by artificial intelligence for targeted database mining. These approaches, coupled with scalable recombinant expression, are poised to expand the binder repertoire essential for any immunoanalytical platform. This growing repository will be key to advancing immunochemical diagnostics across the water cycle. T2 - 5th International Symposium on Nanobody for Immunological Analysis and Application CY - Guangzhou, China DA - 15.11.2025 KW - Antibodies KW - Immunoassay KW - Microfluidics KW - Biosensor KW - Trace substances KW - Electrochemical detection PY - 2025 AN - OPUS4-64923 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schneider, Rudolf T1 - How can antibodies help us in the risk assessment of pharmaceuticals in the environment? N2 - The widespread use of pharmaceuticals has led to their persistent presence in various environmental compartments, raising concerns about ecological and human health risks since they pose a significant threat to non-target organisms and human health, necessitating robust methods for environmental risk assessment.   Traditional analytical methods, such as liquid chromatography-mass spectrometry (LC-MS), offer high sensitivity and specificity but are often resource-intensive, require complex sample preparation, limiting their applicability for large-scale or real-time monitoring. In this context, antibody-based analytical techniques present a promising complementary approach for the detection and quantification of pharmaceutical residues in environmental matrices.   Antibodies, due to their high specificity and affinity, enable the development of immunoassays such as ELISA, lateral flow assays, fluorescence-based formats, and electrochemical sensors that are rapid, cost-effective, and adaptable to field conditions. These methods can be tailored to target a wide range of pharmaceutical compounds, including antibiotics and endocrine disruptors. Moreover, antibody-based assays facilitate high-throughput screening and can be integrated into portable platforms for on-site analysis, thereby enhancing temporal and spatial resolution in environmental monitoring.   This presentation explores the potential of antibodies in environmental risk assessment, focusing on their application in detecting trace levels of pharmaceuticals in water, soil, and biota. They are presented as a complementary and powerful tool for rapid, cost-effective screening in the lab and that also can be deployed for on-site analysis in the field.   The talk also discusses recent advances in antibody engineering, multiplexing capabilities [1], and the integration of immunoassays with sensor technologies [2].   Case studies illustrate how immunoanalytical data can help in finding potential contamination hotspots [3], hazard identification, pollution source tracking, and the generation of crucial data for predictive models on environmental fate and effects. Antibody-based methods can also help in early warning systems and the evaluation of mitigation strategies, contributing to a more comprehensive and proactive risk assessment framework, eventually to regulatory decision-making, ultimately aiding in the protection of ecosystems and public health. T2 - ICRAPHE - 6th International Conference on Risk Assessment of Pharmaceuticals in the Environment CY - Aveiro, Portugal DA - 20.10.2025 KW - Antibodies KW - Immunoassay KW - Microfluidics KW - Electrochemical detection KW - Biosensor PY - 2025 AN - OPUS4-64922 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - INPR A1 - Döring, Sarah A1 - Wulfes, Birte S. A1 - Atanasova, Aleksandra A1 - Jaeger, Carsten A1 - Walzel, Leopold A1 - Tscheuschner, Georg A1 - Flemig, Sabine A1 - Gawlitza, Kornelia A1 - Feldmann, Ines A1 - Konthur, Zoltán A1 - Weller, Michael G. T1 - Corundum Particles as Trypsin Carrier for Efficient Protein Digestion N2 - Reusable enzyme carriers are valuable for proteomic workflows, yet many supports are expensive or lack robustness. This study describes the covalent immobilization of recombinant trypsin on micrometer-sized corundum particles and assesses their performance in protein digestion and antibody analysis. The corundum surface was cleaned with potassium hydroxide, silanized with 3-aminopropyltriethoxysilane and activated with glutaraldehyde. Recombinant trypsin was then attached, and the resulting imines were reduced with sodium cyanoborohydride. Aromatic amino acid analysis (AAAA) estimated an enzyme loading of approximately 1 µg/mg. Non-specific adsorption of human plasma proteins was suppressed by blocking residual aldehydes with a Tris-glycine-lysine buffer. Compared with free trypsin, immobilization shifted the temperature optimum from 50 to 60 °C and greatly improved stability in 1 M guanidinium hydrochloride. Activity remained above 80 % across several reuse cycles, and storage at 4 °C preserved functionality for weeks. When applied to digesting the NISTmAb, immobilized trypsin provided peptide yields and sequence coverage comparable to soluble enzyme and outperformed it at elevated temperatures. MALDI-TOF MS analysis of Herceptin digests yielded fingerprint spectra that correctly identified the antibody and achieved >60 % sequence coverage. The combination of low cost, robustness and analytical performance makes corundum-immobilized trypsin an attractive option for research and routine proteomic workflows. KW - Aluminum oxide KW - Mass spectrometry KW - Enzyme immobilization KW - Antibodies KW - Protein quantification PY - 2025 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-647944 DO - https://doi.org/10.20944/preprints202510.2002.v1 SP - 1 EP - 22 PB - Preprints.org AN - OPUS4-64794 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - THES A1 - Ecke, Alexander T1 - Integrated Diagnostics of Pharmaceutical Contaminants in Water Supply and Management Systems N2 - The contamination of drinking water with pharmaceuticals represents a severe health risk. In order to monitor the drinking water quality continuously and enable quick countermeasures in case of contamination, novel sensors are required. Here, immunoanalytical methods based on the binding of the analyte to highly selective antibodies can be helpful. In this work, magnetic bead-based immunoassays (MBBAs) have been developed for the detection of two relevant contaminants of drinking water: diclofenac (DCF) and amoxicillin (AMX). In case of the latter, not only the parent drug is of interest in the risk assessment but also its hydrolysis products (HPs). In a comprehensive study, the influence of external factors and intrinsic properties of the water on the rate of hydrolysis was investigated. As the hydrolysis of AMX further impacts the recognition by the antibody, a strategy to analyze samples with unknown hydrolysis degree of AMX was established employing the enzyme β-lactamase in sample preparation. For both analytes, the MBBAs enable the fast quantification with results obtained in less than one hour which represents a major improvement over conventional immunoassays like the enzyme-linked immunosorbent assay (ELISA). Compared to the respective ELISAs with the same antibodies, the MBBAs further exhibit improved analytical parameters such as a broader measurement range and lower limits of detection. Due to the magnetic properties of the beads that serve as a platform for the assays, they are suitable for the mobile and automated detection at the point-of-care. An integrated diagnostic system was designed in which electrochemical detection with chronoamperometry on a microfluidic chip allows for further miniaturization of the system to enable monitoring of the drinking water quality online in water supply pipes at waterworks. N2 - Die Kontamination von Trinkwasser mit Arzneimitteln stellt eine ernste Gesundheitsgefahr dar. Um die Trinkwasserqualität kontinuierlich überwachen und im Falle einer Verunreinigung zeitnah reagieren zu können, sind neuartige Sensoren erforderlich. Hier können immunanalytische Methoden, die auf der Bindung des Analyten an hochselektive Antikörper beruhen, hilfreich sein. In dieser Arbeit wurden magnetpartikelbasierte Immunoassays (MBBAs) für zwei relevante Kontaminanten des Trinkwassers entwickelt: Diclofenac (DCF) und Amoxicillin (AMX). Bei letzterem erwiesen sich neben der Ausgangsverbindung auch dessen Hydrolyseprodukte (HPs) als relevant für die Gefährdungsbeurteilung. In einer umfassenden Studie wurde der Einfluss von externen Faktoren und intrinsischen Eigenschaften des Wassers auf die Hydrolysegeschwindigkeit untersucht. Da die Hydrolyse von AMX auch die Erkennung durch den Antikörper beeinflusst, wurde eine Strategie zur Analyse von Proben mit unbekanntem Hydrolysegrad von AMX unter Verwendung des Enzyms β-Lactamase in der Probenvorbereitung entwickelt. Für beide Analyten ermöglichen die MBBAs eine schnelle Quantifizierung mit Ergebnissen in weniger als einer Stunde, was eine wesentliche Verbesserung gegenüber herkömmlichen Immunoassays wie dem Enzyme-linked Immunosorbent Assay (ELISA) darstellt. Im Vergleich zu den entsprechenden ELISAs mit denselben Antikörpern weisen die MBBAs zudem verbesserte analytische Parameter auf, wie einen breiteren Messbereich und niedrigere Nachweisgrenzen. Aufgrund der magnetischen Eigenschaften der Partikel, die als Plattform für die Assays dienen, eignen sie sich für den mobilen und automatisierten Einsatz vor Ort. Ein integriertes Diagnosesystem, bei dem die elektrochemische Detektion mittels Chronoamperometrie auf einem mikrofluidischen Chip eine weitere Miniaturisierung des Systems ermöglicht, wurde entworfen, um die Überwachung der Trinkwasserqualität online in Wasserwerken zu ermöglichen. KW - Antibiotics KW - Immunoassay KW - Amoxicillin KW - Diclofenac KW - Biosensor KW - Antibodies PY - 2023 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-591901 DO - https://doi.org/10.18452/25853 SP - 1 EP - 129 CY - Berlin AN - OPUS4-59190 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - INPR A1 - Paul, Martin A1 - Tannenberg, Robert A1 - Tscheuschner, Georg A1 - Ponader, Marco A1 - Weller, Michael G. T1 - Cocaine detection by a laser-induced immunofluorometric biosensor N2 - The trafficking of illegal drugs by criminal networks at borders, harbors, or airports is an increasing issue in public health as these routes ensure the main supply of illegal drugs. The prevention of drug smuggling, including the installation of scanners and other analytical devices to detect ultra-small traces of drugs within a reasonable time frame, remains a challenge. The presented immunosensor is based on a monolithic affinity column with a large excess of immobilized hapten, which traps fluorescently labeled antibodies as long as the analyte cocaine is absent. In the presence of the drug, some binding sites of the antibody will be blocked, which leads to an immediate breakthrough of the labeled protein, detectable by highly sensitive laser-induced fluorescence with the help of a Peltier-cooled complementary metal-oxide-semiconductor (CMOS) camera. Liquid handling is performed with high-precision syringe pumps and microfluidic chip-based mixing devices and flow cells. The biosensor achieved limits of detection of 23 pM (7 ppt) of cocaine with a response time of 90 seconds and a total assay time below 3 minutes. With surface wipe sampling, the biosensor was able to detect 300 pg of cocaine. This immunosensor belongs to the most sensitive and fastest detectors for cocaine and offers near-continuous analyte measurement. KW - Drug search KW - Customs KW - Confiscation KW - Border surveillance KW - Narcotics KW - International drug trade KW - Drug trafficking KW - Illicit drug KW - Immunosensor KW - Antibodies KW - Detection PY - 2021 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-529951 DO - https://doi.org/10.20944/preprints202107.0521.v1 SN - 2310-287X N1 - Geburtsname von Ponader, Marco: Wilke, M. - Birth name of Ponader, Marco: Wilke, M. SP - 1 PB - MDPI CY - Basel AN - OPUS4-52995 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - INPR A1 - Ponader, Marco A1 - Röder, Bettina A1 - Paul, Martin A1 - Weller, Michael G. T1 - Sintered glass monoliths as new supports for affinity columns N2 - A novel stationary phase for affinity separations is presented. This material is based on sintered borosilicate glass readily available as semi-finished filter plates with defined porosity and surface area. The material shows fast binding kinetics and excellent long-term stability under real application conditions due to lacking macropores and high mechanical rigidity. The glass surface can be easily modified with standard organosilane chemistry to immobilize selective binders or other molecules used for biointeraction. In this paper, the manufacturing of the columns and their respective column holders by 3D printing is shown in detail. The model system protein A/IgG was chosen as an example to examine the properties of such monolithic columns under realistic application conditions. Several specifications, such as (dynamic) IgG capacity, pressure stability, long-term performance, productivity, non-specific binding, and peak shape, are presented. It could be shown that due to the very high separation speed, 250 mg antibody per hour and column can be collected, which surpasses the productivity of most standard columns of the same size. The total IgG capacity of the shown columns is around 4 mg (5.5 mg/mL), which is sufficient for most tasks in research laboratories. The cycle time of an IgG separation can be less than 1 minute. Due to the glass material's excellent pressure resistance, these columns are compatible with standard HPLC systems. This is usually not the case with standard affinity columns, limited to manual use or application in low-pressure systems. The use of a standard HPLC system also improves the ability for automation, which enables the purification of hundreds of cell supernatants in one day. The sharp peak shape of the elution leads to an enrichment effect, which might increase the concentration of IgG by a factor of 3. The final concentration of IgG can be around 7.5 mg/mL without the need for an additional nanofiltration step. The purity of the IgG was > 95% in one step and nearly 99% with a second polishing run. KW - Affinity Chromatography KW - Glass KW - Purification KW - Antibodies KW - Solid support KW - HPLC KW - FPLC KW - Separation PY - 2021 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-529117 DO - https://doi.org/10.20944/preprints202103.0298.v1 SN - 2310-287X N1 - Geburtsname von Ponader, Marco: Wilke, M. - Birth name of Ponader, Marco: Wilke, M. SP - 1 EP - 17 PB - MDPI CY - Basel AN - OPUS4-52911 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -