Screening of ligand binding behavior using a high-throughput method and development of guidelines for a learning material

Sammanfattning: This study is comprised of two distinct parts, a chromatographic study and a didactic study.Chromatographic studyThe need for higher selectivity in chromatography purification has increased in recent years. Multimodal resins, offering novel selectivity, are a possible solution to this demand. The purpose of this study is to develop new multimodal resins with novel selectivity, using an iterative workflow. Therefore, eleven novel multimodal ligands were screened according to binding behavior using a high-throughput (HT) method. The mapping of binding behaviors was comprised of six proteins and 32 different binding buffers, with various salt concentrations and pH-levels, to allow for a wide, but efficient mapping. The data generated from the screenings were presented using binding capacity and partition coefficient and were evaluated against each other using principal component analysis (PCA). The PCA created a ligand diversity map, which separated ligands in respect to binding behavior. The information supplied by the diversity map can be used for selecting ligands for further research. The results from presenting the data in the form of partition coefficient and binding capacity were comparable, which indicated that the effects of initial protein concentrations were low. However, this study found that there are advantages and disadvantages to using both quantities. It is possible that some of the ligands screened in this study will be used in further research and it is likely that the diversity map will facilitate the development of ligands with novel selectivityDidactic studyWith a rising demand for HT screening as a method to evaluate the binding behaviors of novel ligands, in combination with the UN’s goal for quality education for all, the incentives for developing quality educational materials for HT screenings have increased. A first step towards educational materials is to outline common difficulties, thereby creating guidelines for developing educational materials. In this study, guidelines for the development of learning materials for HT screenings have been produced. The guidelines were based on identification of critical steps for the experimental procedure and utilizing employer experience of learning materials in general. Two methods for data collection was used in the didactic study: observations and interviews. The observations were performed during the training of the screening method. For the interviews, three individual and one focus group interview were held. Thereafter, a thematic analysis was performed on the data to generate themes. The results from the observations and interviews indicated that the critical steps of the learning process were the practical parts. The result of the thematic analysis showed the identification of three themes: Cohesion, Facilitation and Interaction. These themes generated four aspirational guidelines: 1. Identify crucial steps of the practical procedure. 2. Adapt to people with different backgrounds. 3. Maintain coherence throughout the learning process. 4. Utilize strategies for interaction, both human interaction and material interaction.