Enhancement of hypermutation frequency in the chicken B cell line DT40 for efficient diversification of the antibody repertoire.

摘要: Abstract Chicken B cell line DT40 continuously accumulates mutations in the immunoglobulin variable region (IgV) gene by conversion and point mutation, both of which are mediated activation-induced cytidine deaminase (AID), thereby producing an antibody (Ab) library that is useful for screening monoclonal Abs (mAbs) vitro. We previously generated engineered named DT40-SW, whose AID expression can be reversibly switched on or off, developed vitro Ab generation system using DT40-SW cells. To efficiently create with sufficient diversity, higher hypermutation frequency advantageous. this end, we a novel DT40-SWΔC, conditionally expresses C-terminus-truncated mutant lacking nuclear export signal. The transcription level DT40-SWΔC cells was similar to wild-type However, protein truncated less than wild type. enriched nuclei cells, although might highly susceptible degradation. In mutation occurred IgV over threefold suggesting lower increase frequency. Thus, may constructing libraries efficient mAbs