Development of a novel epitope mapping system: RIEDL insertion for epitope mapping method

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Pages (from-to)162-167
Number of pages6
JournalMonoclonal antibodies in immunodiagnosis and immunotherapy
Volume40
Issue number4
DOIs
Publication statusPublished - 2021 Aug 1
Externally publishedYes

Keywords

  • C44Mab-5
  • CD44
  • epitope mapping
  • monoclonal antibody
  • RIEDL tag

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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