2024 AIChE Annual Meeting
(531a) Invited Talk: Designing with Nanoscale Building Blocks: How Protein Engineering Enables New Solutions for Medicine, Sustainability, and Materials
Author
Danielle Tullman-Ercek - Presenter, Northwestern University
Self-assembling proteins make up precisely ordered nanostructures from filaments to capsids to transporters, each of which is promising for applications ranging from biomanufacturing to medicine to materials. For example, a nanoscale protein-based virus-like particle could serve as a vaccine scaffold or as a delivery vehicle for cellular or gene therapy. However, such structures must be tunable for each application, and despite great leaps in our ability to predict how amino acid sequences will fold into a soluble protein, it remains a significant challenge to predict how proteins come together to form the assemblies and machines that are ubiquitous to life. To address this challenge, and inspired by advances in next-generation DNA synthesis and sequencing, we developed a workflow to fully characterize the assembly competency of all possible single mutations in several model systems, including those from a virus-like particle, a bacterial organelle, and a secretion system. The resulting high-resolution datasets challenge several conventional protein design assumptions on the composition of linkers, mutability of pores, and more. We then used the same approach but screened for desired functions to enhance the performance of each system in its target application space. For example, a protein filament of a secretion system was engineered to confer >2-fold higher production of a target product, a virus-like particle was engineered for improved endosomal release upon sensing a drop in pH, and a bacterial microcompartment was engineered to produce biochemicals in a sustainable manner. With this talk, I will dive into two examples of how our sequencing-based approach is useful as a tool for uncovering the fundamental rules of self-assembly as well as for engineering new function into self-assembling systems, highlighting how such approaches may be used to generate nanoscale precision design in next-generation materials.