Kim Sang Jick, Hong Hyo Jeong
Immunotherapy Research Center, Korea Research Institute of Bioscience and Biotechnology, Gwahangno, Yuseong-gu, Daejeon, Republic of Korea.
Methods Mol Biol. 2012;907:247-57. doi: 10.1007/978-1-61779-974-7_14.
Guided selection provides a powerful tool for humanization of the preexisting nonhuman antibodies as exemplified by HUMIRA, the world's first human antibody approved. This chapter describes the sequential guided selection procedure in which mouse VL and VH domains are replaced sequentially with human VL and VH, respectively to derive completely human antibody. The detailed protocols for construction of phage-displayed antibody library, panning, screening, and characterization, are included to achieve successful selection of human antibody with similar characteristics to original mouse antibody.
定向筛选为将已有的非人源抗体人源化提供了一种强大的工具，如全球首个获批的人源抗体修美乐（HUMIRA）所示。本章描述了顺序定向筛选程序，其中小鼠VL和VH结构域分别依次被人源VL和VH取代，以获得完全人源化抗体。文中还包括构建噬菌体展示抗体文库、淘选、筛选和鉴定的详细方案，以成功筛选出与原始小鼠抗体具有相似特性的人源抗体。
