. Author manuscript; available in PMC: 2022 Feb 19.

Published in final edited form as: ACS Chem Biol. 2021 Jan 22;16(2):344–359. doi: 10.1021/acschembio.0c00865

Table 3.

Binding affinities of AzF- and OPG-containing variants of Donkey1.1 and Bovine2.2 clones. Affinities were determined from binding titrations with biotinylated IgGs using a saturation binding model on GraphPad Prism Version 8.3.0. Fold change from WT indicates the effect of ncAA incorporation on binding interactions between the scFv and IgGs (see Materials and Methods for details).

Clone	aaRS/ncAA	Target Antigen	K_D (95% CI), nM	Fold Change from WT $(\frac{K_{D (T A G)}}{K_{D (W T)}})$
Donkey1.1 L93TAG	AcFRS/AzF	Donkey IgG	125 (99–158)	14.3
Donkey1.1 L93TAG	LeuOmeRS/OPG		45.0 (29.2–68.5)	5.10
Donkey1.1 H31TAG	AcFRS/AzF		561 (517–610)	64.1
Donkey1.1 H31TAG	LeuOmeRS/OPG		365 (335–399)	41.7
Donkey1.1 H54TAG	AcFRS/AzF		213 (173–263)	24.4
Donkey1.1 H54TAG	LeuOmeRS/OPG		55.8 (38.8–79.8)	6.40
Bovine2.2 L93TAG	AcFRS/AzF	Bovine IgG	332 (273–406)	7.20
Bovine2.2 L93TAG	LeuOmeRS/OPG		179 (155–208)	3.90
Bovine2.2 H31TAG	AcFRS/AzF		546 (474–633)	11.9
Bovine2.2 H31TAG	LeuOmeRS/OPG		601 (515–708)	13.0
Bovine2.2 H54TAG	AcFRS/AzF		380 (310–471)	8.30
Bovine2.2 H54TAG	LeuOmeRS/OPG		183 (132–254)	4.00