RRC ID 68153
著者 Asano T, Kaneko MK, Kato Y.
タイトル Development of a Novel Epitope Mapping System: RIEDL Insertion for Epitope Mapping Method.
ジャーナル Monoclon Antib Immunodiagn Immunother
Abstract To clarify the binding region of monoclonal antibodies (mAbs) to target molecules, it is very essential to understand the pharmacological function of each mAb. Although deletion mutants and point mutants are usefully utilized for epitope mapping, we often experience the difficulty of determining the mAb epitope against membrane proteins. We aimed to develop a novel method to determine the binding region of mAbs using epitope tag system. We first checked the reactivity of an anti-CD44 mAb (C44Mab-5) to several deletion mutants of CD44. We then employed the RIEDL tag system ("RIEDL" peptide and LpMab-7 mAb). We inserted the "RIEDL" peptide into the CD44 protein from the 21st to 41st amino acid (AA). The transfectants produced were stained by LpMab-7 and C44Mab-5 in flow cytometry. C44Mab-5 did not react with 30th-361st AA of the deletion mutant of CD44. Furthermore, the reaction of C44Mab-5 to RIEDL tag-inserted CD44 from 25th to 36th AA was lost, although LpMab-7 detected most of the RIEDL tag-inserted CD44 from 21st to 41st AA. The epitope of C44Mab-5 for CD44 was determined to be the peptide from 25th to 36th AA of CD44 using RIEDL insertion for epitope mapping (REMAP) method. The REMAP method might be useful for determining the critical epitope of functional mAbs against many target molecules.
巻・号 40(4)
ページ 162-167
公開日 2021-8-1
DOI 10.1089/mab.2021.0023
PMID 34424761
MeSH Antibodies, Monoclonal* Epitope Mapping Epitopes / genetics Flow Cytometry
リソース情報
遺伝子材料 Genome Network Project Human cDNA Clone IRAL013P10 (HGY085570)