TY - JOUR 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. N2 - Wiederverwendbare Enzymcarrier sind für proteomische Arbeitsabläufe von großem Wert, doch viele Trägermaterialien sind teuer oder nicht robust genug. Diese Studie beschreibt die kovalente Immobilisierung von rekombinantem Trypsin auf mikrometergroßen Korundpartikeln und bewertet deren Leistungsfähigkeit beim Proteinverdau und der Antikörperanalyse. Die Korundoberfläche wurde mit Kaliumhydroxid gereinigt, mit 3-Aminopropyltriethoxysilan silanisiert und mit Glutaraldehyd aktiviert. Anschließend wurde rekombinantes Trypsin gebunden, und die entstandenen Imine wurden mit Natriumcyanoborhydrid reduziert. Die aromatische Aminosäureanalyse (AAAA) ergab eine geschätzte Enzymbeladung von etwa 1 µg/mg. Die unspezifische Adsorption von menschlichen Plasmaproteinen wurde durch Blockierung der restlichen Aldehyde mit einem Tris-Glycin-Lysin-Puffer unterdrückt. Im Vergleich zu freiem Trypsin verschob die Immobilisierung das Temperaturoptimum von 50 auf 60 °C und verbesserte die Stabilität in 1 M Guanidiniumhydrochlorid erheblich. Die Aktivität blieb über mehrere Wiederverwendungszyklen hinweg bei über 80 %, und die Lagerung bei 4 °C bewahrte die Funktionalität über Wochen hinweg. Beim Verdau des NISTmAb lieferte das immobilisierte Trypsin Peptidausbeuten und Sequenzabdeckungen, die mit denen des löslichen Enzyms vergleichbar waren, und übertraf dieses bei erhöhten Temperaturen. Die MALDI-TOF-MS-Analyse von Herceptin-Verdauen ergab Fingerabdruck-Spektren, die den Antikörper korrekt identifizierten und eine Sequenzabdeckung von 60 % erreichten. Die Kombination aus niedrigen Kosten, Robustheit und analytischer Leistungsfähigkeit macht auf Korund immobilisiertes Trypsin zu einer attraktiven Option für die Forschung und routinemäßige proteomische Arbeitsabläufe. KW - Bottom-up proteomics KW - Solid-phase digestion KW - Biopharmaceutical analysis KW - Peptide mapping KW - Non-specific binding KW - amino acid analysis PY - 2026 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-657021 DO - https://doi.org/10.3390/biotech15010002 SN - 2673-6284 VL - 15 IS - 1 SP - 1 EP - 23 PB - MDPI AG CY - Basle, Switzerland AN - OPUS4-65702 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Oberbeckmann, Sonja T1 - Microplastic-microbiome interactions in aquatic ecosystems: a complex story N2 - As pervasive and persistent pollutants, microplastics exist alongside diverse microbial communities in nature. These hardy substrates accumulate rich polymeric matrices from their surroundings that in turn offer nutrition and protection to diverse communities and their theatre of activity, representing a new ecological niche. With our work, we aim to establish a holistic understanding of the interactions between microplastics and microorganisms within aquatic systems, and how these interactions shape their surrounding environment. In our research, we combine field sampling with in situ experiments, and apply high-throughput sequencing technologies to 16S genes and whole bacterial genomes. We demonstrate through our studies that biofilm communities exhibit no specificity to plastic polymer type but appear rather to be opportunistic colonizers more strongly shaped by spatial and temporal factors. While the polymer surface does not appear to elicit a selective effect on associated biofilms, our studies show that microplastics might shape communities in a different way. Based on our detection of carotenoid synthesis pathways and photosynthetic gene clusters among plastic biofilms, we postulate this new niche to shape a specific functional toolkit, adaptive among its members as a product of prolonged exposure to UV radiation at the sea’s surface. To fully assess the interactions between microplastics and microorganisms, however, one cannot consider plastics singularly but as a component within a more complex and interconnected system. It is known, for instance, that plastics can sorb significant concentrations of polycyclic aromatic hydrocarbons (PAH) in the environment. We show, in our work, that the PAH sorption patterns of microplastics correlate significantly with the structure of associated biofilm communities. Elevated concentrations of specific PAHs on microplastics coincided with the enrichment of selected taxa reportedly capable of hydrocarbon utilisation as well as a reduced diversity among associated biofilm communities. Here, we present a synthesis of our collective research on microplastic biofilms in the aquatic environment, the factors that shape them, and their interactions with nature. To fully comprehend how microplastics impact ecosystems, the entire substrate must be considered, including all chemicals integrated into the polymeric matrix as well as associated biofilms. T2 - SETAC Europe 35th Annual Meeting CY - Vienna, Austria DA - 11.05.2025 KW - Microplastics KW - Biofilms KW - PAH PY - 2025 AN - OPUS4-63154 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Oberbeckmann, Sonja T1 - The microplastic microbiome – an example for the intersection of biology and material research N2 - Biofilms live on the interface between human-made materials and the environment. Using aquatic plastics as an example, we explore their composition, their interactions with the plastic substrate, and their potentially useful functional roles. The presentation will further discuss the importance of considering plastic-associated pollutants in the investigation of such biofilms. T2 - Microbiological Colloquium ICBM CY - Oldenburg, Germany DA - 10.07.2025 KW - Microplastics KW - Microbiome KW - Biofilm PY - 2025 AN - OPUS4-63836 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Song, Jessica T1 - Close encounters on a micro scale: Dynamics and interactions of microplastic biofilms in aquatic ecosystems. N2 - Microplastics circulate freely throughout aquatic ecosystems and, due to their interactive nature, accumulate complex polymeric matrices consisting of rich organic compounds and inorganic pollutants. Simultaneously, these hardy substrates offer nutrition and protection to diverse microbial communities and their theatre of activity, representing a new ecological niche. In our work, we investigate the interactions of microplastics in aquatic systems and characterise the effects of these interactions on associated microbial communities to better understand how these substrates might impact surrounding ecosystems. Demonstrating no specificity to polymer type, microplastic biofilms are shaped more by the strong influence of spatial and temporal factors. Microplastic sorption of polycyclic aromatic hydrocarbons (PAHs), in contrast, appear to be more strongly dictated by substrate type, with different polymers observed to sorb varying levels of different PAHs. These interactions between the different emerging contaminants were found in our study to have a significant effect on associated substrate biofilms. Elevated levels of specific 3- and 4-ring PAHs on polyethylene and polystyrene were found to coincide with a notable shift in community composition and structure, as well as a reduced diversity among biofilm communities. The findings in our study illustrate the importance of investigating the collective effect of pollutants in combination and their complex interactions in assessing their environmental impact. To fully understand how microplastics interact and alter surrounding ecosystems, the entire substrate must be considered, including all chemicals integrated into the polymeric matrix. T2 - Biofilms 11 Conference CY - Cardiff, Wales, UK DA - 13.05.2025 KW - Microplastics KW - Biofilms KW - PAHs PY - 2025 AN - OPUS4-63389 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Piechotta, Christian T1 - Development, Validation and Application of an Accelerated Weathering Protocol for Assessing Bisphenol A Release from Polycarbonate Materials into the environment N2 - Bisphenol A (BPA) is under continuous regulatory scrutiny and listed as substance of very high concern (SVHC). Particular concern is related to its frequent detection in surface waters, despite being readily biodegradable. Several studies have been conducted to investigate sources and pathways of BPA in the environment, concluding that its main use as monomer in polycarbonate (PC) contributes only marginally to environmental BPA releases over its life cycle. To better understand the actual releases of BPA from PC under environmental conditions, a newly developed methodology (Federal Institute for Materials Research and Testing, BAM) was applied, which comprises a novel accelerated weathering protocol for polycarbonate (PC) materials, combined with an advanced analytical setup allowing for improved detection of BPA at trace level concentrations. The weathering protocol achieves a 13.6-fold acceleration compared to Central European outdoor conditions and simulates environmental stressors (global radiation, rain, temperature variations) in a laboratory weathering chamber, with simultaneous BPA release measurements using an organic isotope dilution calibration LC-MS/MS approach. Validation was performed in parallel outdoor exposure tests, by using haze and yellowness index measurements as reference parameters. PC sample types representative of major polycarbonate applications were examined: Samples with different levels of UV-protection as used in transparent sheets used outdoor in construction or housings in Electro- and Electronic applications as well as samples with a protective coating as used in automotive applications (headlamps, glazing and construction). Results demonstrated consistently low total BPA releases of around 0.3 mg m⁻² for samples with exposed PC surfaces, while releases from coated samples were significantly reduced by around two orders of magnitude. In all cases examined, the BPA release ceased to zero after a period of four to six weeks, equivalent to approximately 1 to 1.5 years of outdoor exposure. This suggests that potential BPA releases diminish to virtually zero after that timespan. While the newly developed test method is not suitable for routine laboratory implementation, it provides crucial quantitative data on BPA release from PC materials during accelerated environmental weathering. T2 - Umwelt 2025 CY - Dessau, Germany DA - 22.09.2025 KW - Bisphenol A KW - Polycarbonate KW - Weathering PY - 2025 AN - OPUS4-64664 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Piechotta, Christian T1 - Development, Validation and Application of an Accelerated Weathering Protocol for Assessing Bisphenol A Release from Polycarbonate Materials into the environment N2 - Bisphenol A (BPA) is under continuous regulatory scrutiny and listed as substance of very high concern (SVHC). Particular concern is related to its frequent detection in surface waters, despite being readily biodegradable. Several studies have been conducted to investigate sources and pathways of BPA in the environment, concluding that its main use as monomer in polycarbonate (PC) contributes only marginally to environmental BPA releases over its life cycle. To better understand the actual releases of BPA from PC under environmental conditions, a newly developed methodology (Federal Institute for Materials Research and Testing, BAM) was applied, which comprises a novel accelerated weathering protocol for polycarbonate (PC) materials, combined with an advanced analytical setup allowing for improved detection of BPA at trace level concentrations. The weathering protocol achieves a 13.6-fold acceleration compared to Central European outdoor conditions and simulates environmental stressors (global radiation, rain, temperature variations) in a laboratory weathering chamber, with simultaneous BPA release measurements using an organic isotope dilution calibration LC-MS/MS approach. Validation was performed in parallel outdoor exposure tests, by using haze and yellowness index measurements as reference parameters. PC sample types representative of major polycarbonate applications were examined: Samples with different levels of UV-protection as used in transparent sheets used outdoor in construction or housings in Electro- and Electronic applications as well as samples with a protective coating as used in automotive applications (headlamps, glazing and construction). Results demonstrated consistently low total BPA releases of around 0.3 mg m⁻² for samples with exposed PC surfaces, while releases from coated samples were significantly reduced by around two orders of magnitude. In all cases examined, the BPA release ceased to zero after a period of four to six weeks, equivalent to approximately 1 to 1.5 years of outdoor exposure. This suggests that potential BPA releases diminish to virtually zero after that timespan. While the newly developed test method is not suitable for routine laboratory implementation, it provides crucial quantitative data on BPA release from PC materials during accelerated environmental weathering. T2 - Umwelt 2025 CY - Dessau, Germany DA - 22.09.2025 KW - Bisphenol A KW - Polycarbonate KW - Weathering PY - 2025 AN - OPUS4-64665 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Bresch, Harald T1 - Nanoparticle Characterisation - The long way to standardisation N2 - Diese Präsentation gibt einen Überblick über die Entwicklung der Nanopartikelforschung von ca. 2005 bis heute. Beginnend mit den Besonderheiten von Nanopartikeln und der Aufnahme in den menschlichen Körper über Messmethoden bis hin zur Entwicklung einer Prüfrichtlinie im Rahmen der OECD und einem Ausblick über die absehbaren digitalen Entwicklungen. T2 - Abteilungsseminar der Abteilung 4 CY - Berlin, Germany DA - 27.02.2025 KW - Nanomaterials KW - Nano KW - OECD KW - Standardisierung KW - Advanced Materials PY - 2025 AN - OPUS4-64977 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Bresch, Harald T1 - The digital product passport N2 - This presentation contains information for industry and the scientific communitiy about the new digital product passport and the demands from the EU. Furthermore a short overview is given about the different supporting activities which are currently under developement by BAM. T2 - 3. Netzwerktag Cluster Nanotechnologie CY - Würzburg, Germany DA - 02.07.2025 KW - DPP KW - ESPR KW - Product Passport KW - Ökodesignrichtlinie KW - DMP PY - 2025 AN - OPUS4-64964 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Frelih, Milena T1 - Material-Microbiome Interactions N2 - Estimations show that the anthropogenic mass, the technosphere, already exceeds the biomass on Earth. Understanding the technosphere-environment interactions and their impacts is key to safe and healthy ecosystems and human societies. At BAM, we aim at promoting awareness for microorganisms living and evolving in contact with human-made materials and technical systems. We are creating datasets of biofilms on plastic and other anthropogenic materials and in the context of important technical challenges (e.g. microbially influenced corrosion or biocide-induced antimicrobial resistance). This enables us to identify and examine key organisms in the investigated systems that potentially harbor new, also industrially relevant, species and traits. Further, we explore material effects on aquatic microbial communities, aiming at the development and establishment of innovative and environmentally relevant methodologies. By using bioinformatic and molecular tools and interdisciplinary research, we enhance the understanding of material-microbiome interactions for a safe and sustainable future. T2 - NFDI4Microbiota Annual Conference CY - Cologne, Germany DA - 30.09.2025 KW - Microbiome KW - Microplastics KW - Black fungi KW - Nanoparticles PY - 2025 AN - OPUS4-64637 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 -