Plant cell walls are composed of an interlinked network of polysaccharides, glycoproteins and phenolic polymers. When addressing the diverse polysaccharides in green plants, including land plants and the ancestral green algae, there are significant overlaps in the cell wall structures. Yet, there are noteworthy differences in the less evolved species of algae as compared to land plants. The dynamic process orchestrating the deposition of these biopolymers both in algae and higher plants, is complex and highly heterogeneous, yet immensely important for the development and differentiation of the cell. However, our understanding of the evolutionary mechanisms, biosynthesis and remodelling is limited, especially due to a lack of sufficient glycomic tools for studying green plants. This poses a serious hindrance for understanding the fundamental processes behind terrestrialisation and vascularisation of green algae, during the development into land plants. Hence, there is a pressing need for rethinking the glycomic toolbox, by developing new and high-throughput (HTP) technology, in order to acquire information of the location and relative abundance of diverse cell wall polymers.

In this dissertation, we describe the development and optimization of HTP screening tools, with the specific aim to define the binding profile of novel molecular probes. In addition, we explore the potential in rethinking the way molecular probes are developed. The results presented in this dissertation describe the binding profile - in more or less high resolution - of two small molecular probes, 11 carbohydrate binding modules and 24 monoclonal antibodies. This was made possible by combining the HTP multiplexing capacity of carbohydrate microarrays with diverse glycomic tools, to downstream characterize the specificity of these molecular probes. This reveals an unprecedented expansion of the panel of well-defined molecular probes for cell wall studies.

We envisage that this development of novel molecular probes, along with the HTP retrospective mapping of binding profiles could contribute to the evolutionary understanding of green plants, thus aiding their industrial applicability, as well as the fundamental biological understanding