Directed evolution of anti-HER2 DARPins by SNAP display reveals stability/function trade-offs in the selection process.

ABSTRACT: In vitro display technologies have proved to be powerful tools for obtaining high-affinity protein binders. We recently described SNAP display, an entirely in vitro DNA display system that uses the SNAP-tag to link protein with its encoding DNA in water-in-oil emulsions. Here, we apply SNAP display for the affinity maturation of a designed ankyrin repeat proteins (DARPin) that binds to the extracellular domain of HER2 previously isolated by ribosome display. After four SNAP display selection cycles, proteins that bound specifically to HER2 in vitro, with dissociation constants in the low- to sub-nanomolar range, were isolated. In vitro affinities of the panel of evolved DARPins directly correlated with the fluorescence intensities of evolved DARPins bound to HER2 on a breast cancer cell li

SUBMITTER: Houlihan G 

PROVIDER: S-EPMC4550541 | biostudies-literature | 2015 Sep

REPOSITORIES: biostudies-literature