Towards a generic prototyping approach for therapeutically-relevant peptides and proteins in a cell-free translation system.

ABSTRACT: Advances in peptide and protein therapeutics increased the need for rapid and cost-effective polypeptide prototyping. While in vitro translation systems are well suited for fast and multiplexed polypeptide prototyping, they suffer from misfolding, aggregation and disulfide-bond scrambling of the translated products. Here we propose that efficient folding of in vitro produced disulfide-rich peptides and proteins can be achieved if performed in an aggregation-free and thermodynamically controlled folding environment. To this end, we modify an E. coli-based in vitro translation system to allow co-translational capture of translated products by affinity matrix. This process reduces protein aggregation and enables productive oxidative folding and recycling of misfolded states under thermodynami

SUBMITTER: Wu Y 

PROVIDER: S-EPMC8752827 | biostudies-literature | 2022 Jan

REPOSITORIES: biostudies-literature