(1309-B) High-Throughput Optimization of Paper-Based Cell-Free Biosensors
Monday, February 5, 2024
2:00 PM – 3:00 PM EST
Location: Exhibit Halls AB
Abstract: Cell-free biosensors ability to be lyophilized, stored, and used on-demand have made cell-free systems into an an ideal tool for biological diagnostics. The addition of paper matrices, however, limits the versatility of cell-free systems that enable high-throughput experimentation as individual tickets need to be made, assembled for single experiments, and analyzed using custom often irreproducible hardware. Here, we present a high-throughput 384-well paper ticket format leveraging acoustic liquid handling to dispense reactions and machine vision to quantify reaction performance from colorimetric and luminescent reporters. The throughput enabled by this technique shifts the bottleneck from experimental execution to selecting the experiments; we therefore implement design-of-experiments to optimize the information gained from each design-build-test-learn cycle. We used these approaches to first optimize the performance of a low-cost CFE formulation that was initially non-functional when embedded in paper; then optimized a nucleic acid sensor for heat-stability; and finally developed a novel nanobody-based sensor for insecticides implicated in colony-collapse disorder. This work showcases the ability to rapidly optimize cell-free biosensors and their value as a modular platform for rapid production of useful diagnostics.