biostudies-literatureUnknown119(1)Groff DRecent advances in cell-free protein synthesis have enabled the folding and assembly of full-length antibodies at high titers with extracts from prokaryotic cells. Coupled with the facile engineering of the Escherichia coli translation machinery, E. coli based in vitro protein synthesis reactions have emerged as a leading source of IgG molecules with nonnatural amino acids incorporated at specific locations for producing homogeneous antibody-drug conjugates (ADCs). While this has been demonstrated with extract produced in batch fermentation mode, continuous extract fermentation would facilitate supplying material for large-scale manufacturing of protein therapeutics. To accomplish this, the IgG-folding chaperones DsbC and FkpA, and orthogonal tRNA for nonnatural amino acid production were Biotechnology and bioengineering162-175https://www.ebi.ac.uk/biostudies/studies/S-EPMC9297987biostudies-literatureDevelopment of an E. coli strain for cell-free ADC manufacturing.PMC9297987Hanson JALiang SLi XYin GGroff DZhou SHallam TJCarlos NAChen RArmstrong SSteiner AfalseDevelopment of an E. coli strain for cell-free ADC manufacturing.Recent advances in cell-free protein synthesis have enabled the folding and assembly of full-length antibodies at high titers with extracts from prokaryotic cells. Coupled with the facile engineering of the Escherichia coli translation machinery, E. coli based in vitro protein synthesis reactions have emerged as a leading source of IgG molecules with nonnatural amino acids incorporated at specific locations for producing homogeneous antibody-drug conjugates (ADCs). While this has been demonstrated with extract produced in batch fermentation mode, continuous extract fermentation would facilitate supplying material for large-scale manufacturing of protein therapeutics. To accomplish this, the IgG-folding chaperones DsbC and FkpA, and orthogonal tRNA for nonnatural amino acid production were 2022-01-01T00:00:00Z2022 Jan2025-05-29T20:45:45.395Z2024-11-07T08:38:31.589ZS-EPMC92979873465522910.1002/bit.27961