Extended Data Fig. 8: Evolution of an anti-GFP nanobody from a computationally-designed 200,000-member naïve nanobody library encoded on AHEAD.
From: Rapid generation of potent antibodies by autonomous hypermutation in yeast
a, Representative FACS plots showing enrichment of a GFP-binding clone from the nanobody library and subsequent emergence and fixation of a mutation that increases GFP binding across AHEAD cycles. b, Affinity (EC50) of the AHEAD-evolved anti-GFP nanobody, NbG1i1, isolated from AHEAD cycle 6 as compared to its parent, NbG1, that fixed in AHEAD cycle 3. Binding of yeast-displayed nanobodies by each concentration of GFP was determined in relicate (n = 3, error bars represent ± s.d.) and EC50s were determined by fitting each binding curve.
