SpringerLink
Log in

Off-target activity of TNF-α inhibitors characterized by protein biochips

  • Original Paper
  • Published:
Analytical and Bioanalytical Chemistry Aims and scope Submit manuscript

Abstract

Tumor necrosis factor-alpha inhibitors are widely and successfully used to treat rheumatic diseases. However, significant side effects have been reported. To detect the potential off-target activities of such inhibitors we characterized two therapeutic antibodies (adalimumab, infliximab) and one receptor fusion protein (etanercept) on protein biochips (UNIchip® AV-400) containing a printed serial dilution of tumor necrosis factor-alpha and about 384 different human proteins. Etanercept binds to ten proteins (affinity: 20–33% of tumor necrosis factor-alpha recognition), and six of these proteins are related to ribosomal proteins. Interestingly, adalimumab binds to the same six proteins related to ribosomal proteins (affinity: 12–18%) as well as to four proteins crucially involved in ribosomal protein synthesis. Alignment of protein sequences indicates no significant sequence homology between these ten proteins bound by the biological drugs with the highest off-target activities. Taken together, our in vitro results demonstrate that a significant number of proteins are recognized by tumor necrosis factor-alpha inhibitors and are related to ribosome biogenesis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

Abbreviations

JIA:

Juvenile idiopathic arthritis

OTA:

Off-target activity

RA:

Rheumatoid arthritis

RPL:

Ribosomal protein (large subunit)

RPS:

Ribosomal protein (small subunit)

SLE:

Systemic lupus erythematosus

w/v:

Weight per volume

References

  1. Walter G, Büssow K, Cahill D, Lueking A, Lehrach H (2000) Protein arrays for gene expression and molecular interaction screening. Curr Opin Microbiol 3:298–302

    Article  CAS  Google Scholar 

  2. Cahill DJ (2001) Protein and antibody arrays and their medical applications. J Immunol Methods 250:81–91

    Article  CAS  Google Scholar 

  3. Lueking A, Possling A, Huber O, Beveridge A, Horn M, Eickhoff H, Schuchardt J, Lehrach H, Cahill DJ (2003) A nonredundant human protein chip for antibody screening and serum profiling. Mol Cell Proteomics 2:1342–1349

    Article  CAS  Google Scholar 

  4. Michaud GA, Salcius M, Zhou F, Bangham R, Bonin J, Guo H, Snyder M, Predki PF, Schweitzer BI (2003) Analyzing antibody specificity with whole proteome microarrays. Nat Biotechnol 21:1509–1512

    Article  CAS  Google Scholar 

  5. Ballow M (2005) -ximab this and -zumab that! Has the magic bullet arrived in the new millennium of medicine and science? J Allergy Clin Immunol 116:738–743

    Article  CAS  Google Scholar 

  6. Scott DL, Kingsley GH (2006) Tumor necrosis factor inhibitors for rheumatoid arthritis. N Engl J Med 355:704–712

    Article  CAS  Google Scholar 

  7. Ellerin T, Rubin RH, Weinblatt ME (2003) Infections and anti-tumor necrosis factor alpha therapy. Arthritis Rheum 48:3013–3022

    Article  CAS  Google Scholar 

  8. van der Klooster JM, Bosman RJ, Oudemans-van Straaten HM, van der Spoel JI, Wester JP, Zandstra DF (2003) Disseminated tuberculosis, pulmonary aspergillosis and cutaneous herpes simplex infection in a patient with infliximab and methotrexate. Intensive Care Med 29:2327–2329

    Article  Google Scholar 

  9. Haerter G, Manfras BJ, de Jong-Hesse Y, Wilts H, Mertens T, Kern P, Schmitt M (2004) Cytomegalovirus retinitis in a patient treated with anti-tumor necrosis factor alpha antibody therapy for rheumatoid arthritis. Clin Infect Dis 39:e88–e94

    Article  Google Scholar 

  10. Gardam MA, Keystone EC, Menzies R, Manners S, Skamene E, Long R, Vinh DC (2003) Anti-tumour necrosis factor agents and tuberculosis risk: mechanisms of action and clinical management. Lancet Infect Dis 3:148–155

    Article  CAS  Google Scholar 

  11. Wallis RS, Ehlers S (2005) Tumor necrosis factor and granuloma biology: explaining the differential infection risk of etanercept and infliximab. Semin Arthritis Rheum 34(Suppl):34–38

    Article  CAS  Google Scholar 

  12. Wallis RS, Broder M, Wong J, Lee A, Hoq L (2005) Reactivation of latent granulomatous infections by infliximab. Clin Infect Dis 41(Suppl):S194–S198

    Article  CAS  Google Scholar 

  13. Ehlers S (2005) Tumor necrosis factor and its blockade in granulomatous infections: differential modes of action of infliximab and etanercept? Clin Infect Dis 41(Suppl):S199–S203

    Article  CAS  Google Scholar 

  14. Maini R, St Clair EW, Breedveld F, Furst D, Kalden J, Weisman M, Smolen J, Emery P, Harriman G, Feldmann M, Lipsky P, Attract Study Group (1999) Infliximab (chimeric anti-tumour necrosis factor alpha monoclonal antibody) versus placebo in rheumatoid arthritis patients receiving concomitant methotrexate: a randomised phase III trial. Lancet 354:1932–1939

    Article  CAS  Google Scholar 

  15. Lipsky PE, van der Heijde DM, St Clair EW, Furst DE, Breedveld FC, Kalden JR, Smolen JS, Weisman M, Emery P, Feldmann M, Harriman GR, Maini RN (2000) Anti-tumor necrosis factor trial in rheumatoid arthritis with concomitant therapy study group. Infliximab and methotrexate in the treatment of rheumatoid arthritis. N Engl J Med 343:1594–1602

    Article  CAS  Google Scholar 

  16. Weinblatt ME, Kremer JM, Bankhurst AD, Bulpitt KJ, Fleischmann RM, Fox RI, Jackson CG, Lange M, Burge DJ (1999) A trial of etanercept, a recombinant tumor necrosis factor receptor: Fc fusion protein, in patients with rheumatoid arthritis receiving methotrexate. N Engl J Med 340:253–259

    Article  CAS  Google Scholar 

  17. Moreland LW, Schiff MH, Baumgartner SW, Tindall EA, Fleischmann RM, Bulpitt KJ, Weaver AL, Keystone EC, Furst DE, Mease PJ, Ruderman EM, Horwitz DA, Arkfeld DG, Garrison L, Burge DJ, Blosch CM, Lange ML, McDonnell ND, Weinblatt ME (1999) Etanercept therapy in rheumatoid arthritis. A randomized, controlled trial. Ann Intern Med 130:478–486

    CAS  Google Scholar 

  18. Moreland LW, Cohen SB, Baumgartner SW, Tindall EA, Bulpitt K, Martin R, Weinblatt M, Taborn J, Weaver A, Burge DJ, Schiff MH (2001) Long-term safety and efficacy of etanercept in patients with rheumatoid arthritis. J Rheumatol 28:1238–1244

    Google Scholar 

  19. Furst DE, Schiff MH, Fleischmann RM, Strand V, Birbara CA, Compagnone D, Fischkoff SA, Chartash EK (2003) Adalimumab, a fully human anti tumor necrosis factor-alpha monoclonal antibody, and concomitant standard antirheumatic therapy for the treatment of rheumatoid arthritis: results of STAR (Safety Trial of Adalimumab in Rheumatoid Arthritis). J Rheumatol 30:2563–2571

    CAS  Google Scholar 

  20. van de Putte LB, Atkins C, Malaise M, Sany J, Russell AS, van Riel PL, Settas L, Bijlsma JW, Todesco S, Dougados M, Nash P, Emery P, Walter N, Kaul M, Fischkoff S, Kupper H (2004) Efficacy and safety of adalimumab as monotherapy in patients with rheumatoid arthritis for whom previous disease modifying antirheumatic drug treatment has failed. Ann Rheum Dis 63:508–516

    Article  Google Scholar 

  21. Targan SR, Hanauer SB, van Deventer SJ, Mayer L, Present DH, Braakman T, DeWoody KL, Schaible TF, Rutgeerts PJ, Crohn’s disease cA2 study group (1997) A short-term study of chimeric monoclonal antibody cA2 to tumor necrosis factor alpha for Crohn’s disease. N Engl J Med 337:1029–1035

    Article  CAS  Google Scholar 

  22. Mease PJ, Goffe BS, Metz J, VanderStoep A, Finck B, Burge DJ (2000) Etanercept in the treatment of psoriatic arthritis and psoriasis: a randomized trial. Lancet 356:385–390

    Article  CAS  Google Scholar 

  23. van der Heijde D, Dijkmans B, Geusens P, Sieper J, DeWoody K, Williamson P, Braun J, Ankylosing Spondylitis Study for the Evaluation of Recombinant Infliximab Therapy Study Group (2005) Efficacy and safety of infliximab in patients with ankylosing spondylitis: results of a randomized, placebo-controlled trial (ASSERT). Arthritis Rheum 52:582–591

    Article  Google Scholar 

  24. Kroesen S, Widmer AF, Tyndall A, Hasler P (2003) Serious bacterial infections in patients with rheumatoid arthritis under anti-TNF-alpha therapy. Rheumatology (Oxford) 42:617–621

    Article  CAS  Google Scholar 

  25. Salliot C, Gossec L, Ruyssen-Witrand A, Luc M, Duclos M, Guignard S, Dougados M (2007) Infections during tumour necrosis factor-{alpha} inhibitor therapy for rheumatic diseases in daily practice: a systematic retrospective study of 709 patients. Rheumatology (Oxford) 46:327–334

    Article  CAS  Google Scholar 

  26. Lueking A, Huber O, Wirths C, Schulte K, Stieler KM, Blume-Peytavi U, Kowald A, Hensel-Wiegel K, Tauber R, Lehrach H, Meyer HE, Cahill DJ (2005) Profiling of alopecia areata auto-antigens based on protein microarray technology. Mol Cell Proteomics 4:1382–1390

    Article  CAS  Google Scholar 

  27. Nicodeme P (2001) Fast approximate motif statistics. J Comput Biol 8(3):235–248

    Article  CAS  Google Scholar 

  28. Gershoni JM (1988) Protein blotting: a manual. Methods Biochem Anal 33:1–58

    Article  CAS  Google Scholar 

  29. Zeng F-Y, Oka JA, Weigel PH (1996) Renaturation and ligand blotting of the major subunit of the rat asialoglycoprotein receptor after denaturing polyacrylamide gel electrophoresis. Glycobiology 6:247–255

    Article  CAS  Google Scholar 

  30. Keane J, Gershon S, Wise RP, Mirabile-Levens E, Kasznica J, Schwieterman WD, Siegel JN, Braun MM (2001) Tuberculosis associated with infliximab, a tumor necrosis factor alpha-neutralizing agent. N Engl J Med 345:1098–1104

    Article  CAS  Google Scholar 

  31. Mohan AK, Cote TR, Block JA, Manadan AM, Siegel JN, Braun MM (2004) Tuberculosis following the use of etanercept, a tumor necrosis factor inhibitor. Clin Infect Dis 39:295–299

    Article  CAS  Google Scholar 

  32. Keystone EC (2005) Safety of biologic therapies—an update. J Rheumatol 74(Suppl):8–12

    Google Scholar 

  33. Wallis RS, Broder MS, Wong JY, Hanson ME, Beenhouwer DO (2004) Granulomatous infectious diseases associated with tumor necrosis factor antagonists. Clin Infect Dis 38:1261–1265

    Article  CAS  Google Scholar 

  34. Nestea MG, Radstake T, Joosten LA, van der Meer JW, Barrera P, Kullberg BJ (2003) Salmonella septicemia in rheumatoid arthritis patients receiving anti-tumor necrosis factor therapy: association with decreased interferon-gamma production and Toll-like receptor 4 expression. Arthritis Rheum 48:1853–1857

    Article  Google Scholar 

  35. Ellerin T, Rubin RH, Weinblatt ME (2003) Infections and anti-tumor necrosis factor alpha therapy. Arthritis Rheum 48:3013–3022

    Article  CAS  Google Scholar 

  36. Crum NF, Lederman ER, Wallace MR (2005) Infections associated with tumor necrosis factor-(alpha) antagonists. Medicine 84:291–302

    Article  CAS  Google Scholar 

  37. Sakanyan V (2005) High-throughput and multiplexed protein array technology: protein-DNA and protein–protein interactions. J Chromatogr B 815:77–95

    Article  CAS  Google Scholar 

Download references

Acknowledgment

We thank Martin Eggert, Uwe Zettl, and Gunther Neeck (University Rostock) for providing adalimumab, etanercept and infliximab. We thank Jens Beator for helpful discussions and the critical review of the manuscript. Axel Kowald gratefully acknowledges funding from the EU and the Ministerium für Innovation, Wissenschaft, Forschung und Technologie des Landes Nordrhein-Westfalen with the project PtJ-Az.: z0511V01. This work was supported by grants from the Forschungskommission der Medizinischen Fakultät der Heinrich Heine Universität Düsseldorf, and by the Elterninitiative Kinderkrebsklinik e.V. Düsseldorf. We thank the Jeffrey Modell Foundation for its support of our Düsseldorf Immunodeficiency Center.

Author information

Authors and Affiliations

  1. Department of Pediatric Oncology, Hematology and Clinical Immunology, Jeffrey Modell Immunodeficiency Center, University Children’s Hospital Düsseldorf, 40225, Düsseldorf, Germany

    Oliver Feyen, Ulrich Göbel & Tim Niehues

  2. Protagen AG, Otto-Hahn-Str.15, 44227, Dortmund, Germany

    Angelika Lueking

  3. Ruhr-University Bochum, Medizinisches Proteom-Center, Universitätsstraße 150, 44780, Bochum, Germany

    Axel Kowald, Christian Stephan & Helmut E. Meyer

  4. Medizinisches Proteom-Center, Ruhr-Universität Bochum, Universitätsstrasse 150, 44780, Bochum, Germany

    Helmut E. Meyer

Authors
  1. Oliver Feyen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Angelika Lueking
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Axel Kowald
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Christian Stephan
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Helmut E. Meyer
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ulrich Göbel
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Tim Niehues
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Helmut E. Meyer.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Feyen, O., Lueking, A., Kowald, A. et al. Off-target activity of TNF-α inhibitors characterized by protein biochips. Anal Bioanal Chem 391, 1713–1720 (2008). https://doi.org/10.1007/s00216-008-1938-7

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00216-008-1938-7

Keywords

Search

Navigation