Cell-Free Protein Synthesis with 2H/15N/13C-Labeled Amino Acids in H2O for the Production of Perdeuterated Proteins with 1H in the Exchangeable Positions

This study illustrates the finding that back‑exchange is redundant if the protein of interest is produced by cell‑free synthesis from perdeuterated amino acids in H2O. Importantly, the costs associated with cell‑free synthesis compare favorably with those of in vivo expression.