earticle

영어

Monoclonal antibodies (mAb) have recently proven to be excellent biopharmaceutical agents. Generation of hybridomas from antigen-stimulated B cells has been a key technology for obtaining mAbs; however, it is a laborious and time-consuming process, and there are sometimes difficulties in acquiring mAbs for molecules conserved between species because of immunological tolerance. Thus, it is of great importance to develop in vitro technologies for generating useful Abs as drug candidates. We have been challenging to develop a novel in vitro antibody generation system using a chicken B cell line DT40, which displays Abs and mutates Ig genes during culture, thereby generating a useful Ab library for screening mAbs. First, we have generated an engineered cell line DT40-SW whose mutation machinery can be reversibly switched on and off. The Abgeneration system using DT40-SW was found to be useful as follows: (1) mAbs for various model antigens including antoantigens can be obtained from DT40-SW Ablibrary that is free from immunological tolerance; (2) the switching device of the mutation
machinery enables to fix desirable Ig mutants by stopping mutation; (3) by repeated culture and sorting of clones bearing higher affinity for target antigens, affinity maturation can be mimicked in vitro. We have also genetically improved DT40-SW cells for mutation efficiency and Abproduction. The Ab generation system will be applicable for obtaining valuable Abs such as anti-tumor Abs.