A Simple Whole-Plasmid PCR Method to Construct High-Diversity Synthetic Phage Display Libraries.

ABSTRACT: Phage display technology utilises peptide and antibody libraries with very high diversities to select ligands with specific binding properties. The production of such libraries can be labour intensive and technically challenging and whilst there are commercial sources of libraries, the exploitation of the resulting binders is constrained by ownership of the libraries. Here, a peptide library of ~ 1 × 10⁹ variants for display on gene VIII was produced alongside three VHH antibody libraries with similar diversity, where 12mer, 16mer or 21mer CDR3s were introduced into the highly stable cAbBCII10 scaffold displayed on gene III. The cloning strategy used a simple whole-plasmid PCR method and type IIS restriction enzyme assembly that facilitate the seamless insertion of diversity int

SUBMITTER: Tsoumpeli MT 

PROVIDER: S-EPMC9217769 | biostudies-literature | 2022 Jul

REPOSITORIES: biostudies-literature