Development and applications of advanced probing tools for cell wall biology

Aleksander Riise Hansen

Abstract

Common to all plant species, the cell wall is the fiber rich tough outer coat that protects the plant cell. This study set out to expand the set of probes against glycans found primarily in the plant cell wall, and explore their application for use in related agroindustrial and fundamental research fields. The first part of the thesis describes the use of an already established method for comprehensive microarray profiling to assess optimal hydrothermal pre-treatment levels and surface properties of composite biomass in important bio-industrial grasses. The method is used to characterize the function of pectin methyl esterase inhibitors and their role in plant defense against microbial degradation, and cell wall structural dynamics in relation to cell detachment from roots. The second part describes phage display as a method for developing probes against targets that are poor immunogens. In this study, antigens consisting of crude mixtures of alkali extracted polymers from the grass model Brachypodium distachyon were targets for probe generation using a naïve human single domain antibody library. Epitope characterization of positive clones from phage ELISA was then further elucidated by printing defined samples of oligosaccharides and polysaccharides using microarray technology to assess specificity and cross reactivity of individual probes

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