2023 AIChE Annual Meeting
Controlling Peptide Binding Chemistry and Density within Gelatin Hydrogels
In an effort to test hydrogels without excessive animal testing, we are developing a microfluidic device to serve as a screening tool. These microfluidic devices will be used to grow vascular tissue in several hydrogel conditions. Each hydrogel has a gelatin base with different vascular promoting peptides chemically bound. My project has focused on synthesizing and quantifying these hydrogels across three different peptide factors: sequence, binding chemistry, and concentration. The three peptide activities being tested include QK (a VEGF mimetic peptide), N-cadherin (cell adhesion peptide), and HepPep (a heparin binding peptide). Two peptide-binding chemistries are being assessed in this experiment, click-chemistry via a maleimide-thiol reaction and EDC/NHS coupling. Finally, each peptide was bound at three concentrations, resulting in a low, medium, and high format of each. In total 18 distinct hydrogel formulations are being created for our screen. Additionally, to accurately quantify the amount of peptide in each dose of hydrogel, all variants will be duplicated with a fluorescently tagged peptide. Utilizing UV-Vis spectroscopy, the quantity of fluorescent peptide within each batch will be measured using Beer Lambertâs law. By combining Nuclear Magnetic Resonance data with the fluorescent dataset, we can accurately quantify the peptide content within each hydrogel dose. Next steps include testing each hydrogel variation in the microfluidic devices to screen with vascular antibodies to determine the best hydrogel formulations. Subsequently, the best formats will progress to mouse fat pad injections for further evaluation and validation.