Fig. 6: The potency of designs can be greatly enhanced by the optimal tandem spacing.
From: A topological refactoring design strategy yields highly stable granulopoietic proteins
The creation of two-copy Boskar4 tandems with different flexible spacers, through A a 24-residue-linker (Boskar4_t2), and B a short 6-residue-linker (Boskar4_st2). C Dose–response curves in NFS-60 cells show Boskar4_t2 (EC50 = 180 pM), and Boskar4_st2 (EC50 = 7.6 pM) to be 11- and 263-fold more active than the single domain Boskar4, respectively. Datapoints and error bars represent mean ± standard deviations of 3 biologically independent replicates. D, E This greatly enhanced potency is also evident in the dose- and time-dependent NFS-60 proliferation kinetics, highlighting the requirement for an optimal G-CSFR dimerisation spacing to achieve maximal G-CSFR activation. The experiments in C, E were performed as described in figure legend to Fig. 2.
