De Novo Isolation & Affinity Maturation of yeast-displayed Virion-binding human fibronectin domains by flow cytometric screening against Virions.

ABSTRACT:

Background

The promise of biopharmaceuticals comprising one or more binding domains motivates the development of novel methods for de novo isolation and affinity maturation of virion-binding domains. Identifying avenues for overcoming the challenges associated with using virions as screening reagents is paramount given the difficulties associated with obtaining high-purity virus-associated proteins that retain the conformation exhibited on the virion surface.

Results

Fluorescence activated cell sorting (FACS) of 1.5 × 10⁷ clones taken from a naïve yeast surface-displayed human fibronectin domain (Fn3) against whole virions yielded two unique binders to Zika virions. Construction and FACS of site-directed binding loop mutant libraries based on one of these binders

SUBMITTER: Heinzelman P 

PROVIDER: S-EPMC6796422 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature