production-optimization-lte-leishmania-tarentolae-derived-cell-free

Production and Optimization of LTE, a <em>Leishmania tarentolae</em> Derived Cell-Free Protein Expression System for Recombinant Protein Production

This protocol outlines the production and optimization of a eukaryotic Cell-Free Protein Expression System (CFPS) derived from the unicellular flagellate ...

Queensland University of Technology

Eukaryotic cell-free protein expression systems enable rapid production of recombinant multidomain proteins in their functional form. A cell-free system based on the rapidly growing protozoan  (LTE) has been extensively used for protein engineering and analysis of protein interaction networks. However, like other eukaryotic cell-free systems, LTE deteriorates at ambient temperatures and requires deep freezing for transport and storage. In this study, we report the development of a lyophilized version of LTE. Use of lyoprotectants such as poly(ethylene glycol) and trehalose during the drying process allows retention of 76% of protein expression activity versus nonlyophilized controls. Lyophilized LTE is capable of withstanding storage at room temperature for over 2 weeks. We demonstrated that upon reconstitution the lyophilized LTE could be used for  expression of active enzymes, analysis of protein-protein interactions by AlphaLISA assay, and functional analysis of protein biosensors. Development of lyophilized LTE lowers the barriers to its distribution and opens the door to its application in remote areas.

Juan Alfaro Palma

CSIRO-QUT Synthetic Biology Alliance,
ARC Centre of Excellence in Synthetic Biology,
Centre for Agriculture and the Bioeconomy,
School of Biology and Environmental Science

Queensland University of Technology

Production and Optimization of LTE, a Leishmania tarentolae Derived Cell-Free Protein Expression System for Recombinant Protein Production

Juan Alfaro Palma

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}CSIRO-QUT Synthetic Biology Alliance,ARC Centre of Excellence in Synthetic Biology,Centre for Agriculture and the Bioeconomy,School of Biology and Environmental Science

Queensland University of Technology

Production and Optimization of LTE, a Leishmania tarentolae Derived Cell-Free Protein Expression System for Recombinant Protein Production

CSIRO-QUT Synthetic Biology Alliance,
ARC Centre of Excellence in Synthetic Biology,
Centre for Agriculture and the Bioeconomy,
School of Biology and Environmental Science