Optical detection of denatured ferritin protein via plasmonic gold nanoparticles exposure through aminosilane solution

Abstract

The presence of denatured proteins within a therapeutic drug product can create a series of serious adverse effects, such as mild irritation, immunogenicity, anaphylaxis, or instant death to a patient. The detection of protein degradation is complicated and expensive due to current methods associated with expensive instrumentation, reagents, and processing time. We have demonstrated here a platform for visual biosensing of denatured proteins that is fast, low cost, sensitive, and user friendly by exploiting the plasmonic properties of noble metal nanoparticles. In this study we have exposed artificially heat stressed ferritin and gold nanoparticles to 3-aminopropyl triethoxysilane, which degrades the protein by showing a systematic blue shift in the absorbance spectra of the gold nanoparticle/ferritin and aminosilane solution. This blue shift in absorbance produces a detectable visual color transition from a blue color to a purple hue. By studying the Raman spectroscopy of the gold nanoparticle/ferritin and aminosilane solution, the extent of ferritin degradation was quantified. The degradation of ferritin was again confirmed using dynamic light scattering and was attributed to the aggregation of the ferritin due to accelerated heat stress. We have successfully demonstrated a proof of concept for visually detecting ferritin from horse spleen that has experienced various levels of degradation, including due to heat stress.