Label-free biomolecular interaction sensing on microarray using surface plasmon resonance imaging

This thesis, “Biomolecular interaction sensing on microarrays using surface plasmon resonance imaging”, gives a brief introduction to biomolecular interactions in general. And includes interactions of peptides and proteins, antibodies and autoimmune diseases. Furhtermore, the concept of label-free biosensing is introduced, with a special emphasis on SPR imaging. The principle and role of microarrays and data analysis are also described. Chapter 3 explains the soft lithography process to develop various types of PDMS spotting devices for the immobilization of proteins in confined surface areas in a microarray format. PDMS was explored to provide a disposable alternative for the TopSpot, however, in combination with our experimental restrictions the PDMS based method did not succeed and we continued using the TopSpot. In chapter 4 and 5 the use of a scanning-angle surface plasmon resonance (SPR) imaging instrument for monitoring the binding of biomolecules on user-defined regions of interest of a microarray is described. Peptides and proteins were both spotted on the same sensor chip to illustrate that both, low and high molecular weight ligands with initial large differences in off-set SPR angles, can be applied within the same experiment. The effectiveness of this system is demonstrated by automatically monitoring the interactions between citrullinated peptides and serum autoantibodies of 50 rheumatoid arthritis (RA) patients and 29 controls in a single step. Chapter 6 deals with a new assay protocol based on injecting a single analyte concentration, and exposing it to spots with various ligand densities, rather than multiple analyte injections of varying analyte concentrations exposed to a single ligand surface, as is done in the conventional procedure is proposed. The new method uses controlled dilution of ligands with background molecules which, facilitates immobilization of a precise ligand density on the surface, which is a prerequisite for direct kinetic analysis. This alternative to the conventional multi-analyte overlay plot for calculating the rate- and affinity constants of a biomolecular interaction, has the advantage of requiring only a single analyte injection, fewer surface regeneration steps, and a reduction of the overall assay time. As a model system, various biotin specific interactions have been tested by means of SPR imaging for their affinity toward surface immobilized biotinylated peptides

Combined Lab-on-a-Chip and microarray approach for biomolecular interaction sensing using surface plasmon resonance imaging

Surface plasmon resonance imaging (SPR) is a well-established label-free detection technique for real-time biomolecular interaction measurements. An integrated LOC sensing system with fluidic control for sample movement to specific locations on microarray surface in combination with SPR imaging is demonstrated by the measurements of human IgG and anti-IgG interactions from 24 patterned regions.\u

Shifting Mindset - From product evolution to experience driven innovation in corporate environments

The BMW Group is one of the world’s leading premium manufacturers of automobiles and motorcycles. However, mobility, together with an increased world population, causes problems like traffic congestions, emissions, and an increased energy and raw material consumption that impacts our climate. This, digital developments like the Internet of Things, and the pressure from emerging car brands like Tesla or new mobility providers, have disrupted the automotive industry. Complexity has therefore increased, creating wicked problems for BMW that has to adapt itself to these dynamic times. It however also creates opportunities opportunities for whole new driving and mobility experiences. To create these experiences, BMW has to rethink its current ways of working that are still largely engineering and product driven. Instead, a more experience driven mindset could be the answer to increasing customer demands and market pressure. This research therefore focuses on developing a strategy on how industries could adapt to dynamic times by changing a company‘s internal mindset, from a product-oriented focus to a more experience driven one. Starting from the experience, means taking a user-centred approach that focusses on human needs instead of merely product- and feature-evolvement. This would not only benefit the customer, but also the company‘s success by generating competitive advantages, profitability and sustaining long-term customer loyalty.Following from the research, it was found that UX and CX can be taken to a next level, which not only optimises customer touchpoints around products and services but solves for human needs around a clear purpose. This is called a Business of Experience (BOX) and the newly framed goal is to create a strategy that transforms the BMW Group into a BOX that is fully human- and experience driven. In line with the 4M framework, the need for four strategies was derived, called CO: strategies, to reach this final goal. On the individual level, a strategy is needed that enhances the engagement of every employee, a.k.a. Individual Contributor (IC), which is translated into CO:NTRIBUTION. On the second level, collaboration and knowledge flow between teams and departments must be enhanced, resulting in a strategy involving CO:LLABORATION. Next to this, there is a need for supporting organisational structures, cultures, and leaders, thus CO:ORDINATION. Lastly, stakeholders that form the community around organisations should be involved in the development process to learn about needs and innovations, and a strategy to enhance CO:-CREATION should be implemented.A roadmap with three horizons and a final vision for the BMW Group was created, that should lead the BMW Group towards becoming a Business of Experience. A good starting point is the creation of a Centre of Excellence (COE). A COE is a centralised and independent, physical and/or virtual facility within an organisation, that leverages the resources that are already there and provides the rest of the organisation with coordination, research, knowledge, support, trainings, and enablers in a specific area. It supports change initiatives, like that of mindset shifts, in corporate environments and constantly explores new tools and practices, which it then transfers across all functions of an organisation.Strategic Product Desig

BAAC rapport V-07.0464

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Angle-scanning SPR imaging for detection of biomolecular interactions on microarrays

In this paper we describe the use of a commercial surface plasmon resonance (SPR) imaging instrument for monitoring the binding of biomolecules on user-defined regions of interest of a microarray. By monitoring the angle shift of the SPR-dip using a continuous angle-scanning mode instead of monitoring the change in reflectivity at a fixed angle, a linear relationship with respect to the mass density change on the surface will remain over a wide dynamic angle range of 8◦. Peptides (2.4 kDa) and proteins (150 kDa) were both spotted on the same sensor chip to illustrate that both, low and high molecular weight ligands with initial large differences in off-set SPR angles, can be applied within the same experiment. By using a fluorescently labeled antibody, SPR results can be confirmed by means of fluorescence microscopy after completion of a SPR experiment. SPR imaging in angle-scanning operation provides sensitive, accurate, and label-free detection of analyte binding on microarrays containing different molecular weight ligands