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

  • Jeremy D. King
  • John C. Williams
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1904)

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).

Key words

Meditope Antibody-drug conjugate Bispecific antibody 

References

  1. 1.
    Strebhardt K, Ullrich A (2008) Paul Ehrlich’s magic bullet concept: 100 years of progress. Nat. Rev. Cancer 8:473.  https://doi.org/10.1038/nrc2394CrossRefPubMedGoogle Scholar
  2. 2.
    Brinkmann U, Kontermann RE (2017) The making of bispecific antibodies. MAbs 9(2):182–212.  https://doi.org/10.1080/19420862.2016.1268307CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Schumacher D, Hackenberger CPR, Leonhardt H et al (2016) Current status: site-specific antibody drug conjugates. J Clin Immunol 36(1):100–107.  https://doi.org/10.1007/s10875-016-0265-6CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Donaldson JM, Zer C, Avery KN et al (2013) Identification and grafting of a unique peptide-binding site in the Fab framework of monoclonal antibodies. Proc Natl Acad Sci U S A 110(43):17456–17461.  https://doi.org/10.1073/pnas.1307309110CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Zer C, Avery KN, Meyer K et al (2017) Engineering a high-affinity peptide binding site into the anti-CEA mAb M5A. Protein Eng Des Sel 30(6):409–417.  https://doi.org/10.1093/protein/gzx016CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    King JD, Ma Y, Kuo Y-C et al (2018) Template-catalyzed, disulfide conjugation of monoclonal antibodies using a natural amino acid tag. Bioconjug Chem 29(6):2074–2081.  https://doi.org/10.1021/acs.bioconjchem.8b00284CrossRefPubMedGoogle Scholar
  7. 7.
    van Rosmalen M, Ni Y, Vervoort DFM et al (2018) Dual-color bioluminescent sensor proteins for therapeutic drug monitoring of antitumor antibodies. Anal Chem 90(5):3592–3599.  https://doi.org/10.1021/acs.analchem.8b00041CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Bzymek KP, Puckett JW, Zer C et al (2018) Mechanically interlocked functionalization of monoclonal antibodies. Nat Commun 9(1):1580.  https://doi.org/10.1038/s41467-018-03976-5CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Molecular MedicineBeckman Research Institute at City of HopeDuarteUSA

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