Synthesis and performance of novel supramolecular tools for single-particle cryogenic electron microscopy and drug and gene delivery

Wright, Kyle J. Ph.D., Purdue University, December 2016. Synthesis and Performance of Novel Supramolecular Tools for Single-Particle Cryogenic Electron Microscopy and Drug and Gene Delivery. Major Professor: David H. Thompson. High-resolution biomacromolecular structure elucidation is fundamentally important to structure-based drug design and basic research into complex biochemical processes. Cryo-EM is an emerging alternative to XRD and NMR that is complementary in many ways relative to XRD and NMR. Materials approaches to cryo-EM are anticipated to greatly facilitate the cryo-EM process, allowing progress toward a more high-throughput application of cryo-EM to address challenges in structural biology. Various affinity-based approaches inspired by approaches previously introduced for 2D crystallization were developed for facilitation of cryo-EM. A library of affinity lipopolymer constructs were synthesized consisting of lipopolymers of various PEG molecular weights conjugated to lipids displaying varied affinity or covalently-active moieties at their distal termini. Modified-EM grids with stabilized lipid coatings were then used to capture His6-T7 bacteriophage and His6-RplL from cell lysates, purified His8-GFPuv, and nanodisc embedded His6-MalFGK2. Graphene-oxide-NTA-based affinity grids were fabricated via covalent modification of graphene-oxide and monolayer