Figure 4

Analytical characterization of PF-06645849. (a) DSC traces of Fc-FGF21 [N171] (solid black line) and Fc-FGF21 [N171][R19V] (solid red line) in PBS. Mid-point transition temperature fits are shown in dashed lines. (b) Western blot analysis of purified PF-06645849 produced in CHO cell lines. Lane 1, post ProteinA elution; lane 2, post ButylFF (GE) flow-through step. (c) Mass spec glycan analysis of PF-06645849 after removal of Fc sugar using IgGZero enzyme. IgGZero hydrolyzes the β1,4 linkage between the core GlcNAc residues in the Fc-glycan, leaving the innermost GlcNAc intact on the Fc (peak labeled 2x). Remaining Glycan structures derived from [N171] are assigned to the corresponding m/z values. (d) pERK phosphorylation assay of WT FGF21, PF-05231023, and HEK- and CHO-produced PF-06645849. Values presented are Mean ± SD (n = 4).