Refining the Quality of Monoclonal Antibodies: Grafting Unique Peptide-Binding Site in the Fab Framework

Methods Mol Biol. 2019:1904:293-298. doi: 10.1007/978-1-4939-8958-4_12.

Abstract

Monoclonal antibodies (mAbs) are a major therapeutic modality. Grafting the meditope binding site onto mAbs, also known as meditope-enabling, can extend the usefulness of mAbs by providing an additional protein-protein interaction surface without altering the stability or antigen binding. We have previously used this site for attaching dyes, cytotoxic drugs, and entire proteins. Here, we provide a simple protocol for meditope-enabling mAbs, and verifying meditope and antigen binding using flow cytometry (FACS).

Keywords: Antibody-drug conjugate; Bispecific antibody; Meditope.

MeSH terms

  • Amino Acid Sequence
  • Antibodies, Monoclonal / chemistry
  • Antibodies, Monoclonal / genetics*
  • Antibodies, Monoclonal / immunology*
  • Binding Sites
  • Computational Biology / methods
  • Humans
  • Immunoglobulin Fab Fragments / chemistry
  • Immunoglobulin Fab Fragments / genetics*
  • Immunoglobulin Fab Fragments / immunology*
  • Mutation
  • Protein Binding / immunology
  • Protein Engineering
  • Structure-Activity Relationship

Substances

  • Antibodies, Monoclonal
  • Immunoglobulin Fab Fragments