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The increasing adoption of Open Science principles has been a prevalent topic in the welding science community over the last years. Providing access to welding knowledge in the form of complex and complete datasets in addition to peer-reviewed publications can be identified as an important step to promote knowledge exchange and cooperation. There exist previous efforts on building data models specifically for fusion welding applications; however, a common agreed upon implementation that is used by the community is still lacking. One proven approach in other domains has been the use of an openly accessible and agreed upon file and data format used for archiving and sharing domain knowledge in the form of experimental data. Going into a similar direction, the welding community faces particular practical, technical, and also ideological challenges that are discussed in this paper. Collaboratively building upon previous work with modern tools and platforms, the authors motivate, propose, and outline the use of a common file format specifically tailored to the needs of the welding research community as a complement to other already established Open Science practices. Successfully establishing a culture of openly accessible research data has the potential to significantly stimulate progress in welding research.
MALDI-TOF-MS-Based Identification of Monoclonal Murine Anti-SARS-CoV-2 Antibodies within One Hour
(2022)
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.