Figure 3 | Scientific Reports

Figure 3

From: RF1 attenuation enables efficient non-natural amino acid incorporation for production of homogeneous antibody drug conjugates

Figure 3

RF1 mutant extract improves nnAA incorporation. (a,b) The amber suppression efficiency increases in HC and LC across most sites in the RF1 mutant extract. (c) The non reducing autoradiography shows the efficient incorporation of pAMF at previously intractable sites using the RF1MUT extract. (d) The reducing autoradiography indicates the suppression efficiency of amber codon in RF1MUT is greatly improved with minimal truncation products. (e) RF1 mutant extract enables incorporation of multiple nnAAs. No increase in truncation prodcts was observed and up to 3nnAAs were incorporated with little loss in expression titer as compared to WT trastuzmab. 1 nnAA = F404, 2 nnAA = F404 + Y180, 3 nnAA = F404 + Y180 + A118, 4 nnAA = F404, Y180, A118, V5, all sites are on HC.

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