Systematic Investigation of Insulin Fibrillation on a Chip
Hoon Suk Rho,
Henk-Willem Veltkamp,
Alexander Thomas Hanke,
Marcel Ottens,
Christian Breukers,
Pamela Habibović,
Han Gardeniers
Affiliations
Hoon Suk Rho
Department of Instructive Biomaterials Engineering, MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht University, 6200 MD Maastricht, The Netherlands
Henk-Willem Veltkamp
Integrated Devices and Systems Group, MESA+ Institute for Nanotechnology, University of Twente, 7522 NB Enschede, The Netherlands
Alexander Thomas Hanke
BioProcess Engineering Group, Department of Biotechnology, Faculty of Applied Sciences, Delft University of Technology, 2628 CD Delft, The Netherlands
Marcel Ottens
BioProcess Engineering Group, Department of Biotechnology, Faculty of Applied Sciences, Delft University of Technology, 2628 CD Delft, The Netherlands
Christian Breukers
Medical Cell BioPhysics Group, Technical Medical Centre, University of Twente, 7522 NB Enschede, The Netherlands
Pamela Habibović
Department of Instructive Biomaterials Engineering, MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht University, 6200 MD Maastricht, The Netherlands
Han Gardeniers
Mesoscale Chemical Systems Group, MESA+ Institute for Nanotechnology, University of Twente, 7522 NB Enschede, The Netherlands
A microfluidic protein aggregation device (microPAD) that allows the user to perform a series of protein incubations with various concentrations of two reagents is demonstrated. The microfluidic device consists of 64 incubation chambers to perform individual incubations of the protein at 64 specific conditions. Parallel processes of metering reagents, stepwise concentration gradient generation, and mixing are achieved simultaneously by pneumatic valves. Fibrillation of bovine insulin was selected to test the device. The effect of insulin and sodium chloride (NaCl) concentration on the formation of fibrillar structures was studied by observing the growth rate of partially folded protein, using the fluorescent marker Thioflavin-T. Moreover, dual gradients of different NaCl and hydrochloric acid (HCl) concentrations were formed, to investigate their interactive roles in the formation of insulin fibrils and spherulites. The chip-system provides a bird’s eye view on protein aggregation, including an overview of the factors that affect the process and their interactions. This microfluidic platform is potentially useful for rapid analysis of the fibrillation of proteins associated with many misfolding-based diseases, such as quantitative and qualitative studies on amyloid growth.
