Tuning The Morphology of Synthetic Bottlebrush Polymers for Protein Structural Determination Using cryoEM

 Dramatic advances over the past decade have occurred in the use of cryogenic electron microscopy (cryoEM) to elucidate the structures of macromolecules at atomic resolution. Unfortunately, the sample preparation process is one of the most time-consuming and empirical methods in the cryoEM workflow. Each sample must be tediously optimized to resolve issues with particle aggregation, ice quality, particle orientation, and particle density to enable high-resolution reconstruction analysis. Post-polymerization modifications of synthetic aqueous bottlebrushes offer a promising approach to streamline the workflow for cryoEM sample preparation. Our approach utilizes synthetic bottlebrush materials comprised of flexible polymer scaffolds bearing grafted side-chains, armed with high affinity ligands at the distal termini of the grafted polymers along the polymer core. Development of water-soluble one-dimensional (1D) synthetic bottlebrush polymers has led to new advancements in the biomaterials, antimicrobial, nanomedicine, and responsive materials fields. These synthetic bottlebrush materials are favorable as they confer properties that linear polymers and small molecules cannot achieve. Moreover, structural manipulations employed during post-polymerization processes can afford bottlebrush polymers with distinguishable topologies for advanced functions. These 1D constructs can be synthesized by atom transfer radical polymerization (ATRP), reversible addition- fragmentation chain-transfer polymerization (RAFT), ring-opening polymerization (ROP), cationic ring-opening polymerization (CROP), anionic ring-opening polymerization (AROP) or ring opening metathesis polymerization (ROMP). The chemical composition of the molecule, number of monomer repeats, grafting density and topology influence the morphology and function of polymer brushes. Elongated, vesicular or micellar morphologies can be specifically tuned for the desired application of the material. The morphology of the polymers can also be manipulated by concentration effects. The morphologies of amphiphilic bottlebrush materials specifically, can typically be influenced by structural topology, solvent choice, or external conditions. ROMP is a living polymerization mechanism that can suffer from catalytic backbiting, causing a loss of livingness. The synthesis of aqueous bottlebrush polymers and the comparison of morphologies via AUC, DLS, AFM and TEM will be presented in this dissertation. The synthetic amphiphilic bottlebrush polymer family presented suffered a loss of livingness and ultimately displayed distinct morphologies, relative to chemical composition, solvent, and ultimately polymerization time. Post-polymerization 11 modifications such as backbone hydrolysis and single-walled carbon nanotube complexation promoted even more unique morphologies of bottlebrushes. These synthetic materials indicate use as promising reagents for cryoEM sample preparation.