Amanda Ferrante

Student: Amanda Ferrante (WINNER)

Project: Protease Identification by Combined Fluorescent Zymography and Proteomics

View: Research Poster (PDF)

Institution: Lehigh University

Major: Bioengineering

Advisor: Thomas Pashuck


Biomaterials, specifically those utilized in tissue engineering, have a variety of applications within the bioengineering field.1 Tissues typically contain multiple different cell types, all working synchronously to achieve some biological function. There is a complex relationship between cell types in both healthy tissues and the regenerative environment desired in biomaterial therapies.2 Each cell type, however, may contain its own specific needs. Therefore, a cellular matrix should contain some sort of targeting mechanism to control each cell type. The Pashuck lab hopes to create a hydrogel matrix by taking advantage of cell-secreted proteases to induce an environmental change and engineered response.3 Our lab is synthesizing protease substrate peptides that are flanked with a fluorophore-quencher pair to track cleavages and an azido-lysine, Fmoc-Lys(N3) to covalently couple the peptides into a polyacrylamide gel. After electrophoretic separation, our lab conducted Liquid Chromatography-Mass Spectrometry (LCMS) to identify the peptides that are present. The cleavages of cell lysates caused a visual increase in fluorescence. After fluorescent bands were excised and processed, a comprehensive LCMS blast was used to identify peptides present. This work presents a promising platform for identifying the proteases that are responsible for peptide cleavages. Once validated, future work includes incorporation of cell-specific protease substrate peptides that are cleaved by unknown proteases. This knowledge can then be applied to cellular targeting within a regenerative environment.
References: 1, Lee, E. J.; Kasper, F. K.; Mikos, A. G. Biomaterials for Tissue Engineering. Ann Biomed Eng 2014, 42 (2), 323–337; 2, Guiseppi‐Elie, A.; Brahim, S. I.; Narinesingh, D. A Chemically Synthesized Artificial Pancreas: Release of Insulin from Glucose-Responsive Hydrogels. Advanced Materials 2002, 14 (10), 743–746; 3, Thomas Pashuck, E. (2018). Synthesis of Self-Assembling Peptide-Based Hydrogels for Regenerative Medicine Using Solid-Phase Peptide Synthesis. In Biomaterials for Tissue Engineering (pp. 177–192). Springer New York. 

About Amanda Ferrante

Amanda Ferrante, a senior bioengineering student at Lehigh University, is a Clare Boothe Luce Research Scholar and undergraduate researcher in the Pashuck Lab. Following graduation, Amanda will be working as a mechanical engineer for Stryker Instruments in Kalamazoo, Michigan. She has performed undergraduate research with Dr. E. Thomas Pashuck since Spring of 2019. The Pashuck Lab is working to create a hydrogel matrix that takes advantage of cell-secreted proteases to facilitate minimally-invasive injectable delivery of cells, regardless of cell type or environment. Primarily, Amanda’s work focuses on the incorporation of fluorescent proteases into a hydrogel to identify the specific proteases that are responsible for peptide cleavages. In addition to bioengineering research, Amanda is a Senior Peer Mentor in the Technical Entrepreneurship Capstone Program, Admissions Fellow, and an Orientation Coordinator for Lehigh’s Class of 2023. Amanda is a great leader on Lehigh’s campus, both inside and outside of the classroom.