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Gewählte Master / Diploma Thesis:

Nina Uhlir (2022): Cell-cell mediated selection of anti-CD59 Fcabs from a yeast display library.
Master / Diploma Thesis - Institut für Pflanzenbiotechnologie und Zellbiologie (IPBT), BOKU-Universität für Bodenkultur, pp 80. UB BOKU obvsg

Data Source: ZID Abstracts
Abstract:
The GPI-anchored CD59 is a member of the membrane-bound complement regulatory proteins (mCRPs) and known for its activity to inhibit the formation of membrane attack complex (MAC) by interacting with C9 and C8α. This mechanism of inhibition protects humoral cells from being collaterally destroyed by their own antibacterial MAC. Many types of cancerogenic cells overexpress CD59 to counter the activation of the complement pathway. Moreover, this molecule is engaged in cell proliferation and metastasis formation and has emerged as a tumor marker, indicating severeness and grade of the disease. This master thesis deals with the development of antigen binding Fc fragment (Fcab) that binds to CD59 and prevents its functioning as a MAC inhibitor. Since CD59 is highly glycosylated, we suggested to replace the selection of yeast Fcab libraries by fluorescence-activated sorting with soluble CD59 with an alternative method, the cell-cell mediated selection, where CD59-expressing cells instead of soluble CD59 are used. We developed a protocol for the staining procedure of the strongly CD59-positive cell lines SK-BR-3 and ARH-77 and used a CD59-enriched yeast Fcab-library for the sorting campaigns. The examination of four potential clones confirmed that the cell-cell mediated selection is a useful method to develop antigen binders with regard to the naturally occurring properties of cell-surface-bound proteins, like glycosylation.

Beurteilende*r: Mach Lukas
1.Mitwirkender: Wozniak-Knopp Gordana

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