(5g) Basic and Applied Studies in Biologically Relevant Polymer Thin Films

Here, we describe a novel approach towards cell and tissue staining based on polymer networks. Our approach exhibits exceptional detail of protein localization at antibody concentrations several orders of magnitude lower than typical immunofluorescent protocols. We will describe our methodology in detail as well as provide a few key results demonstrating a 3 order of magnitude reduction in antibody consumption. We will demonstrate the effectiveness of this approach in the staining of both large regions of cytoplasm and small localized protein structures on the nuclear envelope. We will also discuss characterization of the polymeric labeling system. While this approach would be effective in reducing antibody consumption in labs, it is particularly useful when paired with custom or rare antibodies.

In addition, I will describe the development of glucose oxidase initiated polymeric hydrogel films. An enzymatic polymerization initiator is a facile, noncytotoxic, room temperature approach for biomaterial generation. Glucose oxidase has been shown to possess exceptional stability and activity, making it particularly attractive as a potential polymerization initiator. Biologically, glucose oxidase is used to convert beta-D-glucose into D-glucono-1,5-lactone, and then regenerates FADH2 by the oxidation of molecular oxygen to hydrogen peroxide. We utilize the enzymatic generation of hydrogen peroxide to react with Fe2SO4 (as in Fenton's Reagent) to form hydroxy radicals, which initiate polymerization. We report our investigation into the both the bulk and thin film polymerization kinetics of hydrogel systems including polyethylene glycol diacrylate and hydroxyethylacrylate. We will discuss the concentration dependence of coinitiation and inhibition by the iron species. We will also highlight potential applications including biodetection and cell encapsulation for culture and immunoisolation applications.