Discipline:
Subcategory: Nanoscience
Monique Farrell - Norfolk State University
Co-Author(s): Robert Reaume, Wagneci Hawley, Alexandra Gurrola, Erin Jenrette, Jasmin Flowers, and Aswini. K. Pradhan, Center for Materials Research Norfolk State University, Norfolk, VA
Every year pharmaceutical companies use significant resources to mitigate aggregation of pharmaceutical drug products. Peptides and proteins that have been denatured or degraded can lead to irritation at the site of injection, anaphylaxis, decrease in drug product potency and immune diseases. Current methods to detect aggregation of biological molecules are limited to costly and time consuming processes such as high pressure liquid chromatography, ultrahigh pressure liquid chromatography and SDS-PAGE gels. Aggregation of pharmaceutical drug products can occur during manufacturing, processing, packaging, shipment and storage. Therefore, a facile in solution detection method was evaluated to visually detect denatured glutathione peptides, ferritin protein and insulin, utilizing gold nanoparticle aggregation via 3-Aminopropyltreithoxysilane. The biological analytes were denatured using a 70°C and 80°C water bath to create an accelerated heat stressed environment. The biological analytes, gold nanoparticle and aminosilane solutions were characterized via, UV-Vis spectroscopy, circular dichroism spectroscopy, FTIR spectroscopy, dynamic light scattering and scanning electron microscopy. Captured images and resulting absorbance spectra of the trials demonstrated visual color changes detectable with the human eye as a function of the denaturation time. This work serves as an extended proof of concept for fast in solution detection of proteins and peptides that have experienced heat stress. Future work can include application of this visual detection method to different classes of drugs such as antibody and antibody drug conjugated systems.
Funder Acknowledgement(s): This work is supported by the NSF-CREST (CNBMD) Grant number HRD 1036494 and NSF-IGERT.
Faculty Advisor: Aswini K. Pradhan, apradhan@nsu.edu
Role: Monique Farrell conceived the idea, designed experiments and executed them. Monique Farrell directed three undergraduates (Robert Reaume, Wagneci Hawley and Alexandra Gurrola) in literature review and instrument use in order to contribute to the research.