Rapidly Inducible Yeast Surface Display for Antibody Evolution with OrthoRep

• DOI: 10.1021/acssynbio.4c00370
• Publication Date: 2024-07-25T16:55:35Z
• Authors: Alexandra M. Paulk; Rory L. Williams; Chang C. Liu
• Keywords: Cell Sorting

We recently developed "autonomous hypermutation yeast surface display" (AHEAD), a technology that enables the rapid generation of potent and specific antibodies in yeast. AHEAD pairs display with an error-prone orthogonal DNA replication system (OrthoRep) to continuously rapidly mutate surface-displayed antibodies, thereby enabling enrichment for stronger binding variants through repeated rounds cell growth fluorescence-activated sorting. currently utilizes standard galactose induction drive selective on surface. However, achieving maximal levels can require up 48 h induction. Here we report updated version platform synthetic β-estradiol-induced gene expression regulate find is notably faster both our traditional from nuclear plasmids do not hypermutate. The was fully functional evolution antibodies.