	US 11,685,942 B2
	Methods for making polypeptides including d-amino acids
George M. Church, Brookline, MA (US); and Po-Yi Huang, Boston, MA (US)
Assigned to President and Fellows of Harvard College, Cambridge, MA (US)
Appl. No. 16/81,989
Filed by President and Fellows of Harvard College, Cambridge, MA (US)
PCT Filed Mar. 3, 2017, PCT No. PCT/US2017/020569 § 371(c)(1), (2) Date Sep. 4, 2018, PCT Pub. No. WO2017/151997, PCT Pub. Date Sep. 8, 2017.
Claims priority of provisional application 62/303,425, filed on Mar. 4, 2016.
Prior Publication US 2020/0283817 A1, Sep. 10, 2020
Int. Cl. C12P 21/00 (2006.01)

CPC C12P 21/00 (2013.01)

18 Claims

1. A method of making a polypeptide including one or more D-amino acids comprising

combining protein translation factors including (1) a ribosome, (2) a template encoding the polypeptide, wherein the template encoding the polypeptide includes one or more codons which have been recoded to accept a tRNA attached to a D-amino acid, (3) a plurality of L-amino acids and a plurality of corresponding tRNAs, (4) a plurality of D-amino acids and their corresponding aminoacyl tRNA synthetase or a plurality of tRNAs ligated with a D-amino acid, and (5) elongation factor P in a concentration of 4 to 8 micromolar,

wherein translation of the template encoding the polypeptide occurs to produce the polypeptide including one or more D-amino acids.