Developing a Cell Free Expression System Derived from Human Primary Cells

Rao, G., University of Maryland Baltimore County
Tolosa, L., University of Maryland Baltimore County
Ge, X., UMBC
Szeto, G., MIT
Kostov, Y., UMBC
Cell free translation is a technology that has traditionally had many applications in aiding in the discovery of many basic science concepts such as the discovery of the codon and reconstructing the mechanisms of translation. Recent advances in cell free translation have broadened the scope of applications to include proteomics and manufacturing. While most cell free protein expression systems are derived from continuous cell lines, one of the first and most well-established eukaryotic cell free translation systems used primary cells harvested from rabbits as a source of raw material. While this system has many merits, there have been limitations identified due to the nature of the cells used as a raw material. Specifically, the Heme Regulated Inhibition (HRI) and tRNA availability were identified as limiting factors as reticulocytes main protein expression function in-vivo is to produce hemoglobin. While producing a human equivalent to this system would be impossible due to ethical constraints, there are still other human derived primary cells which could be used to produce a cell free translation system. Here we report the development of a cell free translation system derived from human peripheral blood mononuclear cells (PBMCs).