A cell-free expression and purification process for rapid production of protein biologics

Challise J. Sullivan, Erik D. Pendleton, Henri H. Sasmor, William L. Hicks, John B. Farnum, Machiko Muto, Eric M. Amendt, Jennifer A. Schoborg, Rey W. Martin, Lauren G. Clark, Mark J. Anderson, Alaksh Choudhury, Raffaella Fior, Yu Hwa Lo, Richard H. Griffey, Stephen A. Chappell, Michael C. Jewett, Vincent P. Mauro, John Dresios*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

38 Scopus citations

Abstract

Cell-free protein synthesis has emerged as a powerful technology for rapid and efficient protein production. Cell-free methods are also amenable to automation and such systems have been extensively used for high-throughput protein production and screening; however, current fluidic systems are not adequate for manufacturing protein biopharmaceuticals. In this work, we report on the initial development of a fluidic process for rapid end-to-end production of recombinant protein biologics. This process incorporates a bioreactor module that can be used with eukaryotic or prokaryotic lysates that are programmed for combined transcription/translation of an engineered DNA template encoding for specific protein targets. Purification of the cell-free expressed product occurs through a series of protein separation modules that are configurable for process-specific isolation of different proteins. Using this approach, we demonstrate production of two bioactive human protein therapeutics, erythropoietin and granulocyte-macrophage colony-stimulating factor, in yeast and bacterial extracts, respectively, each within 24 hours. This process is flexible, scalable and amenable to automation for rapid production at the point-of-need of proteins with significant pharmaceutical, medical, or biotechnological value.

Original languageEnglish (US)
Pages (from-to)238-248
Number of pages11
JournalBiotechnology Journal
Volume11
Issue number2
DOIs
StatePublished - Feb 1 2016

Keywords

  • Cell-free protein synthesis
  • Granulocyte-macrophage colony-stimulating factor
  • Protein Biologics
  • Protein expression

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Molecular Medicine

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