Epitope Mapping of an Anti-Human Epidermal Growth Factor Receptor Monoclonal Antibody (EMab-51) Using the RIEDL Insertion for Epitope Mapping Method

Monoclon Antib Immunodiagn Immunother. 2021 Aug;40(4):149-155. doi: 10.1089/mab.2021.0010.ABSTRACTThe classic method for identifying the epitope that monoclonal antibodies (mAbs) bind uses deletion mutants and point mutants of the target protein. However, determining the epitope of mAbs-reactive membrane proteins is often challenging. We recently developed the RIEDL insertion for epitope mapping (REMAP) method to identify mAb-binding epitopes. Herein, we first checked the reactivity of an anti-epidermal growth factor receptor (EGFR) mAb (EMab-51) to several EGFR deletion mutants such as EGFR/dN152, EGFR/dN313, EGFR/dN370, EGFR/dN375, EGFR/dN380, and EGFR/dN482. We found the N-terminus of the EMab-51-binding epitope between residues 375 and 380 of EGFR. We next produced EGFR/dN313 mutants with the RIEDL peptide tag inserted at each possible position of 375-AFRGDSFTHTPPLDP-389. EMab-51 lost its reactivity with the mutants having a RIEDL tag inserted at each position of 377-RGDSFTHTPP-386, whereas LpMab-7 (an anti-RIEDL mAb) detected every mutant. Thus, using the REMAP method, we identified the EMab-51-binding epitope of EGFR as 377-RGDSFTHTPP-386.PMID:34424763 | DOI:10.1089/mab.2021.0010

Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - August 23, 2021 Category: Microbiology Authors: Ren Nanamiya Masato Sano Teizo Asano Miyuki Yanaka Takuro Nakamura Masaki Saito Tomohiro Tanaka Hideki Hosono Nami Tateyama Mika K Kaneko Yukinari Kato Source Type: research