Article
RF1 attenuation enables efficient non-natural amino acid incorporation for production of homogeneous antibody drug conjugates.
Scientific reports - 8 Jun 2017
Yin Gang, Stephenson Heather T, Yang Junhao, Li Xiaofan, Armstrong Stephanie M, Heibeck Tyler H, Tran Cuong, Masikat Mary Rose, Zhou Sihong, Stafford Ryan L, Yam Alice Y, Lee John, Steiner Alexander R, Gill Avinash, Penta Kalyani, Pollitt Sonia, Baliga Ramesh, Murray Christopher J, Thanos Christopher D, McEvoy Leslie M, Sato Aaron K, Hallam Trevor J
Abstract excerpt
Amber codon suppression for the insertion of non-natural amino acids (nnAAs) is limited by competition with release factor 1 (RF1). Here we describe the genome engineering of a RF1 mutant strain that enhances suppression efficiency during cell-free protein synthesis, without significantly impacting cell growth during biomass production. Specifically, an out membrane protease (OmpT) cleavage site was engineered...
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