Generation of a Large Peptide Phage Display Library by Self-Ligation of Whole-Plasmid PCR Product

• DOI: 10.1021/acschembio.0c00497
• Publication Date: 2020-10-30T20:51:05Z
• Authors: Xu-Dong Kong; Vanessa Carle; Cristina Díaz-Perlas; Kaycie Butler; Christian Heinis
• Keywords: Library; Panning (audio); Primer (cosmetics); Phagemid

The success of phage display, used for developing target-specific binders based on peptides and proteins, depends the size diversity library screened, but generating large libraries phage-encoded polypeptides remains challenging. New peptide display developed in recent years rarely contained more than 1 billion clones, which appears to have become upper limit generated with reasonable effort. Here, we established a strategy whole-plasmid PCR self-ligation clone 2 × 1010 members. enormous could be obtained through amplifying entire vector DNA by PCR, omitted step isolation from bacterial cells, appending coding via primer, enabled efficient circularization end-ligation facilitate difficult vector-insertion fragments. Panning repertoires against target yielded high-affinity ligands validated quality thus new cloning strategy. This simple places larger within reach nonspecialist researchers hopefully expand possible targets applications.