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

Research output: Contribution to journalArticleScientificpeer-review

5 Citations (Scopus)
51 Downloads (Pure)

Abstract

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.

Original languageEnglish
Article number1380
JournalMolecules
Volume25
Issue number6
DOIs
Publication statusPublished - 2020

Keywords

  • Dual concentration gradients
  • High-throughput screening
  • Insulin fibrillation
  • Microfluidics

Fingerprint

Dive into the research topics of 'Systematic investigation of insulin fibrillation on a chip'. Together they form a unique fingerprint.

Cite this