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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.
Identity management is at the forefront of applications’ security posture. It separates the unauthorised user from the legitimate individual. Identity management models have evolved from the isolated to the centralised paradigm and identity federations. Within this advancement, the identity provider emerged as a trusted third party that holds a powerful position. Allen postulated the novel self-sovereign identity paradigm to establish a new balance. Thus, extensive research is required to comprehend its virtues and limitations. Analysing the new paradigm, initially, we investigate the blockchain-based self-sovereign identity concept structurally. Moreover, we examine trust requirements in this context by reference to patterns. These shapes comprise major entities linked by a decentralised identity provider. By comparison to the traditional models, we conclude that trust in credential management and authentication is removed. Trust-enhancing attribute aggregation based on multiple attribute providers provokes a further trust shift. Subsequently, we formalise attribute assurance trust modelling by a metaframework. It encompasses the attestation and trust network as well as the trust decision process, including the trust function, as central components. A secure attribute assurance trust model depends on the security of the trust function. The trust function should consider high trust values and several attribute authorities. Furthermore, we evaluate classification, conceptual study, practical analysis and simulation as assessment strategies of trust models. For realising trust-enhancing attribute aggregation, we propose a probabilistic approach. The method exerts the principle characteristics of correctness and validity. These values are combined for one provider and subsequently for multiple issuers. We embed this trust function in a model within the self-sovereign identity ecosystem. To practically apply the trust function and solve several challenges for the service provider that arise from adopting self-sovereign identity solutions, we conceptualise and implement an identity broker. The mediator applies a component-based architecture to abstract from a single solution. Standard identity and access management protocols build the interface for applications. We can conclude that the broker’s usage at the side of the service provider does not undermine self-sovereign principles, but fosters the advancement of the ecosystem. The identity broker is applied to sample web applications with distinct attribute requirements to showcase usefulness for authentication and attribute-based access control within a case study.
Nation, migration, narration
(2022)
En France et en Allemagne, l’immigration est devenue dans les dernières décennies une problématique centrale. C’est dans ce contexte qu’est apparu le rap. Celui-ci connaît une popularité énorme chez les populations issues de l’immigration. Pour autant, les rappeurs ne s’en confrontent pas moins à leur identité française ou allemande.
Le but de ce travail est d’expliquer cette apparente contradiction : comment des personnes issues de l’immigration, exprimant un mal-être face à un racisme qu’ils considèrent omniprésent, peuvent-elles se sentir pleinement françaises / allemandes ?
On a divisé le travail entre les chapitres suivants : Contexte de l'étude, méthodologie et théories (I) ; Analyse des différentes formes d’identité nationale au prisme du corpus (II) ; Analyse en trois étapes chronologiques du rapport à la société dans les textes des rappeurs (III-V) ; étude de cas de Kery James en France et Samy Deluxe en Allemagne (VI).