In the present paper, we describe a novel approach to map monoclonal antibody epitopes, using three new monoclonal antibodies that recognize h-TG2 (human transglutaminase 2) as an example. The target gene was fragmented and cloned upstream of an antibiotic-resistance gene, in the vector pPAO2, to select for in-frame polypeptides. After removal of the antibiotic-resistance gene by Cre/Lox recombination, an antigen fragment phage display library was created and selected against specific monoclonal antibodies. Using the h-TG2 fragment library, we were able to identify epitopes. This technique can also be broadly applied to the study of protein–protein interactions.
Characterizing monoclonal antibody epitopes by filtered gene fragment phage display
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Roberto DI NIRO, Fortunato FERRARA, Tarcisio NOT, Andrew R. M. BRADBURY, Fernando CHIRDO, Roberto MARZARI, Daniele SBLATTERO; Characterizing monoclonal antibody epitopes by filtered gene fragment phage display. Biochem J 15 June 2005; 388 (3): 889–894. doi: https://doi.org/10.1042/BJ20041983
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