HUMANIZATION AND AFFINITY-OPTIMIZATION OF ANTIBODIES

摘要

In an efficient selection of higher affinity binders for an antibody humanization strategy, original CDRs are grafted, but no homology modeling is required to determine the key residues in a framework region. One human variable region heavy chain or "VH' and one variable region light chain or 'VL" germline segments show characteristics of high expression in E. coli as a universal template. Key residues in the framework scaffold were determined by screening of available antibody crystal structure data as well as published documents that described the refinements of a specific mouse humanization project. Diversified residues that represent either human antibody selections or mouse selections were integrated into those key framework positions that were assumed to be important in either supporting CDRs or affecting the overall domain structure. A phage display Fab library was used to screen functional reshaped antibodies, which facilitates the isolation of well-maintained clones in a standard panning.