2020 Virtual AIChE Annual Meeting
(182d) Automated mRNA Display for Rapid, Affordable, Target-Directed Reagent Development
Authors
To automate mRNA display, we converted the cyclic process into a network of functional microfluidic components using programmable equipment. The microfluidic components are fabricated using a bench-top 3D printer, which enables straightforward, rapid, and low-cost prototyping of the devices throughout the system development process. For instance, we designed a microfluidic enrichment device (MFED) for automating the bead-based purification steps in mRNA display. In bead-based purifications, peptides are allowed to bind the bead-immobilized target and the ones that are not bound to the target are removed. When performed manually, the removal of unbound peptides is done by repeatedly washing the beads using centrifugal filters. This approach, however, is not applicable to automation. Instead, we automated the purification steps using the MFED which immobilizes the target-bead complexes within a microfluidic channel. The device enables automated bead washing in a continuous-flow manner.
To benchmark the performance of the automated system, we performed controlled enrichments using previously prepared ligands against B-cell lymphoma-extra large (Bcl-xL), a pro-survival cancer protein. We demonstrated that the MFED considerably outperforms the manual technique, with higher enrichment capability per round of selection. Using the MFED, the automated system can further accelerate mRNA display and lower the cost to generate reagents because of its superior performance in selection.
The automated mRNA display has transformative potential in addressing the current challenges of reagent development. By combining microfluidic automation and mRNA display, we aim to create an inexpensive platform for ligand discovery, expand the use of reagents in the biomedical and biopharma community, and thereby accelerate the evolution of cancer diagnostic and therapeutic tools.