Biofunctionalization of Water-Soluble poly(Phenylene Ethynylene)s

We report the use of amide coupling chemistry to covalently link five different biofunctional groups onto an anionic water-soluble poly(phenylene ethynylene) (PPE) polymer. Two of the biofunctionalized PPEs are used in prototype applications, including pH sensing and flow cytometry labeling. The PPE is functionalized with carboxylate (R-CO2-) and sulfonate (R-SO3-) ionic groups. By using an activated ester, the amine-functionalized groups are covalently linked to the PPE polymer via amide linkages. The reaction chemistry is optimized using biotin-ethylene diamine, making it possible to control the loading of the biotin functionality on the PPE chains. Using the optimized approach, a family of five PPEs were prepared that contain biotin, rhodamine, cholesterol, mannose, or folic acid moieties appended to the polymer backbones. The rhodamine- and biotin-modified PPEs were further applied for pH response and flow cytometry applications. The reported approach can be utilized for other classes of water-soluble conjugated polymers, allowing facile development of a variety of new functionalized water-soluble conjugated polymers for a range of applications including sensing, bioimaging, and flow cytometry analysis.