Advertisement

Investigation of Antibody-Drug Conjugates by Mass Spectrometry

  • Madhuri Jayathirtha
  • Costel C. DarieEmail author
Chapter
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 1140)

Abstract

ADCs are empowered monoclonal antibodies that are designed to harness their targeting ability by linking them to cell-killing agents. They are made up of three main components, the antibody, linker and the cytotoxic drug. The specificity of the antibody with the antigen on the tumor cell surface helps with its internalization into the cell after which the active drug is released causing cell death. The investigation of ADCs can be done using a variety of MS methods. Here, we talk about the bottom-up approach, the top-down approaches such as ECD and ETD, the ESI/MS method and IM-MS. Further, we also focus on the applications of MALDI/MS such as UV-MALDI, IR-MALDI and IMS-MALDI and provide examples of the mass spectra that provide tremendous amount of information on ADC structures.

Keywords

Mass spectrometry Antibody-drug conjugates Protein characterization Therapeutics 

Abbreviations

ADC

Antibody-drug conjugates

ALCL

Anaplastic large cell lymphoma

CDRs

Complementary determining regions

DAR

Drug to antibody ratio

ECD

Electron capture dissociation

ESI

Electrospray ionization

ETD

Electron transfer dissociation

HL

Hodgkin’s lymphoma

IM

Ion mobility

mAbs

Monoclonal antibodies

MALDI

Matrix assisted laser desorption ionization

MMAE

Monomethyl auristatin E

MS

Mass spectrometry

PTM

Posttranslational modification

SCLC

Small cell lung carcinoma

TDCs

THIOMAB drug conjugates

TOF

Time of flight

Notes

Acknowledgements

We would like to thank the past and current lab members for creating a pleasant working environment.

References

  1. 1.
    Gorovits, B., et al. (2013). Bioanalysis of antibody-drug conjugates: American Association of Pharmaceutical Scientists Antibody-Drug Conjugate Working Group position paper. Bioanalysis, 5(9), 997–1006.CrossRefGoogle Scholar
  2. 2.
    Diamantis, N., & Banerji, U. (2016). Antibody-drug conjugates--An emerging class of cancer treatment. British Journal of Cancer, 114(4), 362–367.CrossRefGoogle Scholar
  3. 3.
    Glennie, M. J., & van de Winkel, J. G. (2003). Renaissance of cancer therapeutic antibodies. Drug Discovery Today, 8(11), 503–510.CrossRefGoogle Scholar
  4. 4.
    Shefet-Carasso, L., & Benhar, I. (2015). Antibody-targeted drugs and drug resistance—Challenges and solutions. Drug Resistance Updates, 18, 36–46.CrossRefGoogle Scholar
  5. 5.
    Adair, J. R., et al. (2012). Antibody–drug conjugates – A perfect synergy. Expert Opinion on Biological Therapy, 12(9), 1191–1206.CrossRefGoogle Scholar
  6. 6.
    Chari, R. V. J. (2008). Targeted cancer therapy: Conferring specificity to cytotoxic drugs. Accounts of Chemical Research, 41(1), 98–107.CrossRefGoogle Scholar
  7. 7.
    Flygare, J. A., Pillow, T. H., & Aristoff, P. (2013). Antibody-drug conjugates for the treatment of cancer. Chemical Biology & Drug Design, 81(1), 113–121.CrossRefGoogle Scholar
  8. 8.
    Erickson, H. K., et al. (2010). Tumor delivery and in vivo processing of disulfide-linked and thioether-linked antibody–maytansinoid conjugates. Bioconjugate Chemistry, 21(1), 84–92.CrossRefGoogle Scholar
  9. 9.
    Kigawa, J., et al. (1998). Glutathione concentration may be a useful predictor of response to second-line chemotherapy in patients with ovarian cancer. Cancer, 82(4), 697–702.CrossRefGoogle Scholar
  10. 10.
    Sanderson, R. J., et al. (2005). In vivo drug-linker stability of an anti-CD30 dipeptide-linked auristatin immunoconjugate. Clinical Cancer Research, 11(2 Pt 1), 843–852.PubMedGoogle Scholar
  11. 11.
    Doronina, S. O., et al. (2006). Enhanced activity of monomethylauristatin F through monoclonal antibody delivery: effects of linker technology on efficacy and toxicity. Bioconjugate Chemistry, 17(1), 114–124.CrossRefGoogle Scholar
  12. 12.
    Wang, L., et al. (2005). Structural characterization of the maytansinoid-monoclonal antibody immunoconjugate, huN901-DM1, by mass spectrometry. Protein Science, 14(9), 2436–2446.CrossRefGoogle Scholar
  13. 13.
    Hamblett, K. J., et al. (2004). Effects of drug loading on the antitumor activity of a monoclonal antibody drug conjugate. Clinical Cancer Research, 10(20), 7063–7070.CrossRefGoogle Scholar
  14. 14.
    Su, D., et al. (2018). Modulating antibody-drug conjugate payload metabolism by conjugation site and linker modification. Bioconjugate Chemistry, 29(4), 1155–1167.CrossRefGoogle Scholar
  15. 15.
    Dan, N., et al. (2018). Antibody-drug conjugates for cancer therapy: Chemistry to clinical implications. Pharmaceuticals (Basel, Switzerland), 11(2), 32.CrossRefGoogle Scholar
  16. 16.
    Parslow, C. A., et al. (2016). Antibody–drug conjugates for cancer therapy. Biomedicine, 4(3), E14.Google Scholar
  17. 17.
    Ansell, S. M. (2014). Brentuximab vedotin. Blood, 124(22), 3197.CrossRefGoogle Scholar
  18. 18.
    Mir, S. S., Richter, B. W. M., & Duckett, C. S. (2000). Differential effects of CD30 activation in anaplastic large cell lymphoma and Hodgkin disease cells. Blood, 96(13), 4307.PubMedGoogle Scholar
  19. 19.
    Wahl, A. F., et al. (2002). The anti-CD30 monoclonal antibody SGN-30 promotes growth arrest and DNA fragmentation in vitro and affects antitumor activity in models of Hodgkin’s disease. Cancer Research, 62(13), 3736.PubMedGoogle Scholar
  20. 20.
    Okeley, N. M., et al. (2010). Intracellular activation of SGN-35, a potent anti-CD30 antibody-drug conjugate. Clinical Cancer Research, 16(3), 888.CrossRefGoogle Scholar
  21. 21.
    Sawaki, M. (2014). Trastuzumab emtansine in the treatment of HER2-positive metastatic breast cancer in Japanese patients. Breast Cancer, 6, 37–41.PubMedGoogle Scholar
  22. 22.
    Hudis, C. A. (2007). Trastuzumab--Mechanism of action and use in clinical practice. The New England Journal of Medicine, 357(1), 39–51.CrossRefGoogle Scholar
  23. 23.
    Burris, H. A., et al. (2011). Trastuzumab emtansine (T-DM1): A novel agent for targeting HER2+ breast cancer. Clinical Breast Cancer, 11(5), 275–282.CrossRefGoogle Scholar
  24. 24.
    Sadeghi, S., Olevsky, O., & Hurvitz, S. A. (2014). Profiling and targeting HER2-positive breast cancer using trastuzumab emtansine. Pharmacogenomics and Personalized Medicine, 7, 329–338.PubMedPubMedCentralGoogle Scholar
  25. 25.
    Corrigan, P. A., et al. (2014). Ado-trastuzumab emtansine: A HER2-positive targeted antibody-drug conjugate. The Annals of Pharmacotherapy, 48(11), 1484–1493.CrossRefGoogle Scholar
  26. 26.
    Barok, M., Joensuu, H., & Isola, J. (2014). Trastuzumab emtansine: Mechanisms of action and drug resistance. Breast Cancer Research, 16(2), 209.CrossRefGoogle Scholar
  27. 27.
    Rowe, J. M., & Löwenberg, B. (2013). Gemtuzumab ozogamicin in acute myeloid leukemia: A remarkable saga about an active drug. Blood, 121(24), 4838.CrossRefGoogle Scholar
  28. 28.
    Baron, J., & Wang, E. S. (2018). Gemtuzumab ozogamicin for the treatment of acute myeloid leukemia. Expert Review of Clinical Pharmacology, 11(6), 549–559.CrossRefGoogle Scholar
  29. 29.
    Breccia, M., & Lo-Coco, F. (2011). Gemtuzumab ozogamicin for the treatment of acute promyelocytic leukemia: Mechanisms of action and resistance, safety and efficacy. Expert Opinion on Biological Therapy, 11(2), 225–234.CrossRefGoogle Scholar
  30. 30.
    de Witte, T., & Amadori, S. (2016). The optimal dosing of gemtuzumab ozagamicin: Where to go from here? Haematologica, 101(6), 653–654.CrossRefGoogle Scholar
  31. 31.
    Jager, E., et al. (2011). Targeted drug delivery by gemtuzumab ozogamicin: Mechanism-based mathematical model for treatment strategy improvement and therapy individualization. PLoS One, 6(9), e24265–e24265.CrossRefGoogle Scholar
  32. 32.
    Huang, R. Y. C., & Chen, G. (2016). Characterization of antibody–drug conjugates by mass spectrometry: Advances and future trends. Drug Discovery Today, 21(5), 850–855.CrossRefGoogle Scholar
  33. 33.
    Wakankar, A., et al. (2011). Analytical methods for physicochemical characterization of antibody drug conjugates. MAbs, 3(2), 161–172.CrossRefGoogle Scholar
  34. 34.
    Wagner-Rousset, E., et al. (2014). Antibody-drug conjugate model fast characterization by LC-MS following IdeS proteolytic digestion. MAbs, 6(1), 173–184.CrossRefGoogle Scholar
  35. 35.
    Xu, K., et al. (2011). Characterization of intact antibody–drug conjugates from plasma/serum in vivo by affinity capture capillary liquid chromatography–mass spectrometry. Analytical Biochemistry, 412(1), 56–66.CrossRefGoogle Scholar
  36. 36.
    Chen, G., et al. (2011). Characterization of protein therapeutics by mass spectrometry: Recent developments and future directions. Drug Discovery Today, 16(1), 58–64.CrossRefGoogle Scholar
  37. 37.
    Hunt, D. F., et al. (1986). Protein sequencing by tandem mass spectrometry. Proceedings of the National Academy of Sciences, 83(17), 6233.CrossRefGoogle Scholar
  38. 38.
    Ge, Y., et al. (2002). Top down characterization of larger proteins (45 kDa) by electron capture dissociation mass spectrometry. Journal of the American Chemical Society, 124(4), 672–678.CrossRefGoogle Scholar
  39. 39.
    Syka, J. E. P., et al. (2004). Peptide and protein sequence analysis by electron transfer dissociation mass spectrometry. Proceedings of the National Academy of Sciences of the United States of America, 101(26), 9528.CrossRefGoogle Scholar
  40. 40.
    Cournoyer, J. J., et al. (2005). Deamidation: Differentiation of aspartyl from isoaspartyl products in peptides by electron capture dissociation. Protein Science, 14(2), 452–463.CrossRefGoogle Scholar
  41. 41.
    O’Connor, P. B., et al. (2006). Differentiation of Aspartic and Isoaspartic Acids Using Electron Transfer Dissociation. Journal of the American Society for Mass Spectrometry, 17(1), 15–19.CrossRefGoogle Scholar
  42. 42.
    Hardouin, J. (2007). Protein sequence information by matrix-assisted laser desorption/ionization in-source decay mass spectrometry. Mass Spectrometry Reviews, 26(5), 672–682.CrossRefGoogle Scholar
  43. 43.
    Takayama, M. (2001). In-source decay characteristics of peptides in matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Journal of the American Society for Mass Spectrometry, 12(4), 420–427.CrossRefGoogle Scholar
  44. 44.
    Veronese, F. M., & Pasut, G. (2005). PEGylation, successful approach to drug delivery. Drug Discovery Today, 10(21), 1451–1458.CrossRefGoogle Scholar
  45. 45.
    Siegel, M. M., et al. (1997). Calicheamicin derivatives conjugated to monoclonal antibodies: Determination of loading values and distributions by infrared and UV matrix-assisted laser desorption/ionization mass spectrometry and electrospray ionization mass spectrometry. Analytical Chemistry, 69(14), 2716–2726.CrossRefGoogle Scholar
  46. 46.
    Wang, L., et al. (2005). Structural characterization of a recombinant monoclonal antibody by electrospray time-of-flight mass spectrometry. Pharmaceutical Research, 22(8), 1338–1349.CrossRefGoogle Scholar
  47. 47.
    Quiles, S., et al. (2010). Synthesis and preliminary biological evaluation of high-drug-load paclitaxel-antibody conjugates for tumor-targeted chemotherapy. Journal of Medicinal Chemistry, 53(2), 586–594.CrossRefGoogle Scholar

Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  1. 1.Biochemistry and Proteomics Group, Department of Chemistry and Biomolecular ScienceClarkson UniversityPotsdamUSA

Personalised recommendations