Establishing the Freeze-Dried Cell-Free Protein Synthesis System as a Paradigm for Studying Protein Folding on Earth and in Microgravity

Abstract

As virtually all fundamental processes for biological life are executed by proteins, the study of protein folding and folding kinetics is essential. In order to advance the study of protein folding in resource-limited settings, it is necessary to develop easy to use, inexpensive, and accessible assays to study protein folding, unfolding, stability, and kinetics. Through this research, I sought to develop a low-resource module that explores the parameters of protein folding in an open, cell-free system. The work proceeded in three stages: to establish an experimental set up, adapt said experimental set up for use in educational settings, and finally to lay the groundwork for space application. Here we show that the study of folding kinetics and the structural stability of fluorescent proteins can be achieved through the freeze-dried, cell-free platform called BioBitsⓇ. Combined, this research brings an enhanced study of protein synthesis and folding to the bench, the classroom, and to space.