Epitope mapping of monoclonal antibodies to calreticulin reveals that charged amino acids are essential for antibody binding

Ann Christina Bergmann, Cecilie Kyllesbech, Rimantas Slibinskas, Evaldas Ciplys, Peter Højrup, Nicole Hartwig Trier*, Gunnar Houen

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Calreticulin is a chaperone protein, which is associated with myeloproliferative diseases. In this study, we used resin-bound peptides to characterize two monoclonal antibodies (mAbs) directed to calreticulin, mAb FMC 75 and mAb 16, which both have significantly contributed to understanding the biological function of calreticulin. The antigenicity of the resin-bound peptides was determined by modified enzyme-linked immunosorbent assay. Specific binding was determined to an 8-mer epitope located in the N-terminal (amino acids 34–41) and to a 12-mer peptide located in the C-terminal (amino acids 362–373). Using truncated peptides, the epitopes were identified as TSRWIESK and DEEQRLKEEED for mAb FMC 75 and mAb 16, respectively, where, especially the charged amino acids, were found to have a central role for a stable binding. Further studies indicated that the epitope of mAb FMC 75 is assessable in the oligomeric structure of calreticulin, making this epitope a potential therapeutic target.

Original languageEnglish
Article number31
JournalAntibodies
Volume10
Issue number3
Number of pages13
ISSN2073-4468
DOIs
Publication statusPublished - 2021

Bibliographical note

Publisher Copyright:
© 2021 by the authors. Licensee MDPI, Basel, Switzerland.

Keywords

  • Antibody
  • Calreticulin
  • Enzyme-linked immunosorbent assay
  • Epitope

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