The Intervening Removable Affinity Tag (iRAT) System for the Production of Recombinant Antibody Fragments
重组DNA
抗体
化学
分子生物学
作者
Norimichi Nomura,Yayoi Nomura,Yumi Sato,So Iwata
出处
期刊:Methods in molecular biology日期:2021-01-01
标识
DOI:10.1007/978-1-0716-1126-5_5
摘要
Fv and Fab antibody fragments are versatile co-crystallization partners that aid in the structural determination of otherwise uncrystallizable proteins, including human/mammalian membrane proteins. Accessible methods for the rapid and reliable production of recombinant antibody fragments have been long sought. In this chapter, we describe the concept and protocols of the intervening removable affinity tag (iRAT) system for the efficient production of Fv and Fab fragments in milligram quantities, which are sufficient for structural studies. As an extension of the iRAT system, we also provide a new method for the creation of genetically encoded fluorescent Fab fragments, which are potentially useful as molecular devices in various basic biomedical and clinical procedures, such as immunofluorescence cytometry, bioimaging, and immunodiagnosis.