Freeze-Dried Cell-Free Protein Expression System in Microchambers Toward Point-of-Care Diagnostics

Abstract

This paper reports cell-free protein expression performed by freeze-dried cell-free protein synthesis (CFPS) system in micro-chambers on a Polydimethylsiloxane (PDMS)-Glass microfluidic device. Previously, the CFPS system was freeze-dried on a piece of paper for Point-Of-Care (POC) diagnostics. However, protein expression on paper-based materials is generally non-homogeneous due to its non-homogeneous structure made from cellulose, limiting the quality of diagnostics. Here, we freeze-dried the CFPS system in the micro-chambers on the microfluidic device for the first time. We then validated the functionality of protein expression after rehydration of the CFPS system in the micro-chambers. Using the developed device equipped with the freeze-dried CFPS system, diagnostics of a test sample containing a small molecule, N-acyl-homoserine-lactone (AHL), was demonstrated.