Towards the commercial production of pharmaceutical proteins using cell-free systems

Michael C. Jewett, Kara A. Calhoun, James R. Swartz

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

Energy substrates and nucleotides represent the major reagent costs associated with traditional cell-free protein synthesis systems. In order for cell-free technology to effectively compete with conventional in vivo rDNA protein expression, these costs must be reduced. Two recent breakthroughs have demonstrated that (1) the most efficient arm of cellular energy production, oxidative phosphorylation, has been activated to solve the ATP supply issue in vitro, and (2) cell-free protein synthesis reactions can be fueled by exchanging nucleoside triphosphates (NTPs) with nucleoside monophosphates (NMPs) when 10mM phosphate is supplemented in cell-free reactions utilizing glucose metabolism. Here, we demonstrate the effects of combining these two breakthroughs on the overall system performance. We obtain protein yields of 700 ug/mL of chloramphenical acetyl transferase (CAT) in a 5-hour batch reaction, and we characterize the activated cell-free metabolism with HPLC analysis. These results increase the attractiveness of cell-free protein biosynthesis as a commercial expression technology.

Original languageEnglish (US)
Title of host publication05AIChE
Subtitle of host publication2005 AIChE Annual Meeting and Fall Showcase, Conference Proceedings
Number of pages1
StatePublished - Dec 1 2005
Event05AIChE: 2005 AIChE Annual Meeting and Fall Showcase - Cincinnati, OH, United States
Duration: Oct 30 2005Nov 4 2005

Other

Other05AIChE: 2005 AIChE Annual Meeting and Fall Showcase
CountryUnited States
CityCincinnati, OH
Period10/30/0511/4/05

ASJC Scopus subject areas

  • Engineering(all)

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