Development of cell-free protein synthesis platforms for disulfide bonded proteins.

作者: Aaron R. Goerke , James R. Swartz

DOI: 10.1002/BIT.21567

关键词:

摘要: The use of cell-free protein synthesis (CFPS) for recombinant production is emerging as an important technology. For example, the openness system allows control reaction environment to promote folding disulfide bonded proteins in a rapid and economically feasible format. These advantages make expression systems particularly well suited producing patient specific therapeutic vaccines or antidotes response threats from natural man-made biological agents pharmaceutical that are difficult produce living cells. In this work we assess versatility modern methods, optimize parameters, highlight importance rationally designed plasmid templates mammalian secreted proteins, fusion antibody fragments our E. coli-based CFPS system. Two unique platforms were established by developing standardized extract preparation protocols generic conditions. Generic conditions enabled all express with best yield at 710 microg/mL, fragment 230 vaccine 300 microg/mL; majority correctly folded. Better yields obtained when optimized each protein. Establishing general enhances potential reliably complex products low capital costs very process development timelines.

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