Fig. 2
Construction of libraries for the human ADLib system. a Replacement and insertion of chicken immunoglobulin LC components (open rectangles) into human counterparts (blue rectangles). The endogenous chicken pseudogenes are replaced with designed pseudogenes. b Replacement of the chicken immunoglobulin HC components (open rectangles) with their human counterparts (green rectangles). Designed pseudogenes are inserted downstream of the chicken pseudogene cluster. c Two approaches to construct the human ADLib system. In the “single clone-derived library (SCL)” approach, chicken immunoglobulin genes are replaced by human counterparts, and the designed human pseudogenes are inserted upstream of the VHDHJH region. The immunoglobulin genes of the knocked-in cells are diversified by TSA treatment to enhance GC. In the other approach, “poly-clone-derived libraries (PCLs)”, human PBMC-derived VHDHJH gene libraries are knocked into the VH region of the cells for PCLs. The obtained knocked-in clones are treated by TSA to expand their diversity in a mixed state
