Supplementary Figure S1 from Development of a [⁸⁹Zr]Zr-labeled Human Antibody using a Novel Phage-displayed Human scFv Library

Supplementary Figure 1. Strategy for WashU II phage display library creation. The human antibody repertoires were amplified by PCR from the cDNA of mixed human peripheral blood mononuclear cells and cloned randomly into a phagemid vector. This vector encodes for a 16 aa linker (G4S)3T between the VH and VL domain of the scFv and adds an additional C-terminal 6xHis and Flag tag. First, the VH and VL repertoires were amplified in the first set of PCRs and completed by SfiI/XhoI (VH) or SalI/NotI (VL) restriction sites in the second set of PCRs. The VH repertories were cloned first by electroporation in E. coli TG1 (Lucigen, #60502), followed by cloning of the VLkappa or VL-lambda repertories.