Novel, Inexpensive, and Scalable Amyloid Fibril Formation Method

Abstract

Wheat flour was used as a source of protein for the in vitro synthesis of Amyloid fibrils to develop a novel and inexpensive fabrication method. Amyloid fibrillation was confirmed by Thioflavin T Fluorescence, using confocal microscopy. A morphological study was carried out by transmission electron microscopy (TEM), which revealed the high aspect ratio of the amyloid fibrils formed via a novel process. An application of the amyloid fibers produced by the novel method is shown to be melatonin sensing. Tests showed that the amyloid samples had a measurable color variation dependent on the melatonin concentration. This newly derived process could prove to be a cost-effective tool for future nano-biomaterial applications in commercial and research settings.

Keywords: amyloid; inexpensive; melatonin biosensing; synthesis.

Figure 1

Simple flowchart of an overview of the amyloid synthesis process.

Figure 2

Confocal microscopy images of ThT-dyed amyloid fibrils taken at increasing magnification.

Figure 3

TEM images of amyloid fibrils at increasing magnifications.

Figure 4

Results from the melatonin biosensing experiment including: (a) the melatonin sensing samples 1 min after addition of Congo Red Dye; (b) melatonin sensing samples 10 min after the addition of Congo Red Dye; (c) plot of mean green value vs. natural log of melatonin concentration based on the imaging of samples 1 min after addition of Congo Red Dye; (d) plot of mean grayscale value vs. natural log of melatonin concentration based on imaging of samples 1 min after the addition of Congo Red Dye. Graphs created on MATLAB.