Identification of a Putative Crf Splice Variant and Generation of Recombinant Antibodies for the Specific Detection of Aspergillus fumigatus

BACKGROUND: Aspergillus fumigatus is a common airborne fungal pathogen for humans. It frequently causes an invasive aspergillosis (IA) in immunocompromised patients with poor prognosis. Potent antifungal drugs are very expensive and cause serious adverse effects. Their correct application requires an early and specific diagnosis of IA, which is still not properly achievable. This work aims to a specific detection of A. fumigatus by immunofluorescence and the generation of recombinant antibodies for the detection of A. fumigatus by ELISA. RESULTS: The A. fumigatus antigen Crf2 was isolated from a human patient with proven IA. It is a novel variant of a group of surface proteins (Crf1, Asp f9, Asp f16) which belong to the glycosylhydrolase family. Single chain fragment variables (scFvs) were obtained by phage display from a human naive antibody gene library and an immune antibody gene library generated from a macaque immunized with recombinant Crf2. Two different selection strategies were performed and shown to influence the selection of scFvs recognizing the Crf2 antigen in its native conformation. Using these antibodies, Crf2 was localized in growing hyphae of A. fumigatus but not in spores. In addition, the antibodies allowed differentiation between A. fumigatus and related Aspergillus species or Candida albicans by immunofluorescence microscopy. The scFv antibody clones were further characterized for their affinity, the nature of their epitope, their serum stability and their detection limit of Crf2 in human serum. CONCLUSION: Crf2 and the corresponding recombinant antibodies offer a novel approach for the early diagnostics of IA caused by A. fumigatus

Rise and Fall of an Anti-MUC1 Specific Antibody

So far, human antibodies with good affinity and specificity for MUC1, a transmembrane protein overexpressed on breast cancers and ovarian carcinomas, and thus a promising target for therapy, were very difficult to generate.A human scFv antibody was isolated from an immune library derived from breast cancer patients immunised with MUC1. The anti-MUC1 scFv reacted with tumour cells in more than 80% of 228 tissue sections of mamma carcinoma samples, while showing very low reactivity with a large panel of non-tumour tissues. By mutagenesis and phage display, affinity of scFvs was increased up to 500fold to 5,7×10(-10) M. Half-life in serum was improved from below 1 day to more than 4 weeks and was correlated with the dimerisation tendency of the individual scFvs. The scFv bound to T47D and MCF-7 mammalian cancer cell lines were recloned into the scFv-Fc and IgG format resulting in decrease of affinity of one binder. The IgG variants with the highest affinity were tested in mouse xenograft models using MCF-7 and OVCAR tumour cells. However, the experiments showed no significant decrease in tumour growth or increase in the survival rates. To study the reasons for the failure of the xenograft experiments, ADCC was analysed in vitro using MCF-7 and OVCAR3 target cells, revealing a low ADCC, possibly due to internalisation, as detected for MCF-7 cells.Antibody phage display starting with immune libraries and followed by affinity maturation is a powerful strategy to generate high affinity human antibodies to difficult targets, in this case shown by the creation of a highly specific antibody with subnanomolar affinity to a very small epitope consisting of four amino acids. Despite these "best in class" binding parameters, the therapeutic success of this antibody was prevented by the target biology

Digitalisation of Person-Oriented Services in the Field of Child and Youth Welfare: Design and Evaluation of an Information System Design

Personenbezogene Dienstleistungen in der Kinder- und Jugendhilfe stellen in Deutschland einen erheblichen Anteil der nachgefragten Dienstleistungen sowie einen bedeutenden Wirtschaftszweig dar. Sie zeichnen sich durch die Interaktion verschiedener Individuen innerhalb komplexer sozialer Prozesse aus. Dabei besteht im täglichen Arbeitsalltag die Herausforderung, schwach strukturierte Arbeitsprozesse, informelle Kommunikationsstrukturen sowie Erfahrungswissen so zu integrieren, dass einerseits eine bestmögliche persönliche Unterstützung der Klienten erreicht wird, andererseits individuelle und organisationale Arbeits- und Lernprozesse ermöglicht werden. Trotz der enormen Bedeutung der personenbezogenen Dienstleistungen existiert in diesem Bereich bisher keine adäquate Unterstützung der Arbeit durch dedizierte Informationssysteme, sodass vielfach auf Consumer-IT zurückgegriffen werden muss. Im vorliegenden Beitrag berichten wir daher über den Entwurf und die Evaluation eines branchenspezifischen Informationssystemdesigns zur Unterstützung personennaher Dienstleistungen im Bereich der Kinder- und Jugendhilfe. Zunächst werden in diesem Bereich die aus mehreren Quellen gewonnenen und zusammengeführten Anforderungen überblickartig vorgestellt. Auf deren Grundlage wird schließlich die Gestaltung eines Informationssystems vollzogen. Der Systemvorschlag wird anschließend im Hinblick auf die geschaffenen Potenziale von Praktikern bewertet sowie einer kritischen Reflexion unterzogen. Abschließend werden Potenziale für die weitere Forschung vorgestellt