Journal of Gastroenterology

, Volume 47, Issue 2, pp 136–143

Development of a new immunoassay for the accurate determination of anti-infliximab antibodies in inflammatory bowel disease

  • Hirotsugu Imaeda
  • Akira Andoh
  • Yoshihide Fujiyama
Original Article—Alimentary Tract



The formation of antibodies to infliximab (ATIs) is closely associated with the loss of response to infliximab in patients with inflammatory bowel disease (IBD). We evaluated the clinical utility of a novel method to measure serum ATI levels in the presence of infliximab.


ATI levels were measured using a novel immunoassay and the conventional method in 58 patients with Crohn’s disease (CD) under infliximab maintenance therapy. The serum infliximab trough levels were determined by enzyme-linked immunosorbent assay.


ATIs were detected in 16 out of 58 patients (27.6%) by the new method, but the conventional method detected only 2 patients (3.4%) who had the two highest ATI titers assayed by the new method. The presence of ATIs in the samples positive by the new method but negative by the conventional method was confirmed by Western blot analysis. Western blotting analysis also indicated that the new method could restore the binding capacities of the ATIs whose recognition sites were occupied by free infliximab. In the new method, the addition of infliximab to the samples dose-dependently blocked the detection of ATIs. Patients positive for ATIs had significantly lower serum trough levels of infliximab (P < 0.01) and significantly higher clinical activity scores (P < 0.001) as compared with patients negative for ATI.


The new method makes it possible to measure serum ATI levels in the presence of infliximab. This method is useful for deciding the optimal management strategies for IBD patients with loss of response to infliximab.


Anti-infliximab antibodies IBD Loss of response 


  1. 1.
    Danese S, Colombel JF, Reinisch W, Rutgeerts PJ. Review article: infliximab for Crohn’s disease treatment—shifting therapeutic strategies after 10 years of clinical experience. Aliment Pharmacol Ther. 2011;33:857–69.PubMedCrossRefGoogle Scholar
  2. 2.
    Mayer L. Evolving paradigms in the pathogenesis of IBD. J Gastroenterol. 2010;45:9–16.PubMedCrossRefGoogle Scholar
  3. 3.
    Hanauer SB, Kornbluth AA, Messick J, Rubin DT, Sandborn WJ, Sands BE. Clinical scenarios in IBD: optimizing the use of conventional and biologic agents. Inflamm Bowel Dis. 2010;16(Suppl 1):S1–11.PubMedCrossRefGoogle Scholar
  4. 4.
    Kobori A, Yagi Y, Imaeda H, Ban H, Bamba S, Tsujikawa T, et al. Interleukin-33 expression is specifically enhanced in inflamed mucosa of ulcerative colitis. J Gastroenterol. 2010;45:999–1007.PubMedCrossRefGoogle Scholar
  5. 5.
    Allez M, Vermeire S, Mozziconacci N, Michetti P, Laharie D, Louis E, et al. The efficacy and safety of a third anti-TNF monoclonal antibody in Crohn’s disease after failure of two other anti-TNF antibodies. Aliment Pharmacol Ther. 2010;31:92–101.PubMedCrossRefGoogle Scholar
  6. 6.
    D’Haens GR, Panaccione R, Higgins PD, Vermeire S, Gassull M, Chowers Y, et al. The London Position Statement of the World Congress of Gastroenterology on Biological Therapy for IBD with the European Crohn’s and Colitis Organization: when to start, when to stop, which drug to choose, and how to predict response? Am J Gastroenterol. 2011;106:199–212 (quiz 3).Google Scholar
  7. 7.
    Hanauer SB, Feagan BG, Lichtenstein GR, Mayer LF, Schreiber S, Colombel JF, et al. Maintenance infliximab for Crohn’s disease: the ACCENT I randomised trial. Lancet. 2002;359:1541–9.PubMedCrossRefGoogle Scholar
  8. 8.
    Sands BE, Anderson FH, Bernstein CN, Chey WY, Feagan BG, Fedorak RN, et al. Infliximab maintenance therapy for fistulizing Crohn’s disease. N Engl J Med. 2004;350:876–85.PubMedCrossRefGoogle Scholar
  9. 9.
    Rutgeerts P, Sandborn WJ, Feagan BG, Reinisch W, Olson A, Johanns J, et al. Infliximab for induction and maintenance therapy for ulcerative colitis. N Engl J Med. 2005;353:2462–76.PubMedCrossRefGoogle Scholar
  10. 10.
    Sandborn WJ, Rutgeerts P, Feagan BG, Reinisch W, Olson A, Johanns J, et al. Colectomy rate comparison after treatment of ulcerative colitis with placebo or infliximab. Gastroenterology. 2009;137:1250–60 (quiz 520).Google Scholar
  11. 11.
    Schnitzler F, Fidder H, Ferrante M, Noman M, Arijs I, Van Assche G, et al. Long-term outcome of treatment with infliximab in 614 patients with Crohn’s disease: results from a single-centre cohort. Gut. 2009;58:492–500.PubMedCrossRefGoogle Scholar
  12. 12.
    Van Assche G, Magdelaine-Beuzelin C, D’Haens G, Baert F, Noman M, Vermeire S, et al. Withdrawal of immunosuppression in Crohn’s disease treated with scheduled infliximab maintenance: a randomized trial. Gastroenterology. 2008;134:1861–8.PubMedCrossRefGoogle Scholar
  13. 13.
    Gisbert JP, Panes J. Loss of response and requirement of infliximab dose intensification in Crohn’s disease: a review. Am J Gastroenterol. 2009;104:760–7.PubMedCrossRefGoogle Scholar
  14. 14.
    Yanai H, Hanauer SB. Assessing response and loss of response to biological therapies in IBD. Am J Gastroenterol. 2011;106:685–98.PubMedCrossRefGoogle Scholar
  15. 15.
    Baert F, Noman M, Vermeire S, Van Assche G, DH G, Carbonez A, et al. Influence of immunogenicity on the long-term efficacy of infliximab in Crohn’s disease. N Engl J Med. 2003;348:601–8.PubMedCrossRefGoogle Scholar
  16. 16.
    Farrell RJ, Alsahli M, Jeen YT, Falchuk KR, Peppercorn MA, Michetti P. Intravenous hydrocortisone premedication reduces antibodies to infliximab in Crohn’s disease: a randomized controlled trial. Gastroenterology. 2003;124:917–24.PubMedCrossRefGoogle Scholar
  17. 17.
    Maser EA, Villela R, Silverberg MS, Greenberg GR. Association of trough serum infliximab to clinical outcome after scheduled maintenance treatment for Crohn’s disease. Clin Gastroenterol Hepatol. 2006;4:1248–54.PubMedCrossRefGoogle Scholar
  18. 18.
    Regueiro M, Siemanowski B, Kip KE, Plevy S. Infliximab dose intensification in Crohn’s disease. Inflamm Bowel Dis. 2007;13:1093–9.PubMedCrossRefGoogle Scholar
  19. 19.
    Ainsworth MA, Bendtzen K, Brynskov J. Tumor necrosis factor-alpha binding capacity and anti-infliximab antibodies measured by fluid-phase radioimmunoassays as predictors of clinical efficacy of infliximab in Crohn’s disease. Am J Gastroenterol. 2008;103:944–8.PubMedCrossRefGoogle Scholar
  20. 20.
    Afif W, Loftus EV Jr, Faubion WA, Kane SV, Bruining DH, Hanson KA, et al. Clinical utility of measuring infliximab and human anti-chimeric antibody concentrations in patients with inflammatory bowel disease. Am J Gastroenterol. 2010;105:1133–9.PubMedCrossRefGoogle Scholar
  21. 21.
    Ben-Horin S, Yavzori M, Katz L, Kopylov U, Picard O, Fudim E, et al. The immunogenic part of infliximab is the F(ab’)2, but measuring antibodies to the intact infliximab molecule is more clinically useful. Gut. 2011;60:41–8.PubMedCrossRefGoogle Scholar
  22. 22.
    Bendtzen K, Ainsworth M, Steenholdt C, Thomsen OO, Brynskov J. Individual medicine in inflammatory bowel disease: monitoring bioavailability, pharmacokinetics and immunogenicity of anti-tumour necrosis factor-alpha antibodies. Scand J Gastroenterol. 2009;44:774–81.PubMedCrossRefGoogle Scholar
  23. 23.
    Hanauer SB, Wagner CL, Bala M, Mayer L, Travers S, Diamond RH, et al. Incidence and importance of antibody responses to infliximab after maintenance or episodic treatment in Crohn’s disease. Clin Gastroenterol Hepatol. 2004;2:542–53.PubMedCrossRefGoogle Scholar
  24. 24.
    Wolbink GJ, Vis M, Lems W, Voskuyl AE, de Groot E, Nurmohamed MT, et al. Development of antiinfliximab antibodies and relationship to clinical response in patients with rheumatoid arthritis. Arthritis Rheum. 2006;54:711–5.PubMedCrossRefGoogle Scholar
  25. 25.
    Vermeire S, Noman M, Van Assche G, Baert F, D’Haens G, Rutgeerts P. Effectiveness of concomitant immunosuppressive therapy in suppressing the formation of antibodies to infliximab in Crohn’s disease. Gut. 2007;56:1226–31.PubMedCrossRefGoogle Scholar
  26. 26.
    Best WR, Becktel JM, Singleton JW, Kern F Jr. Development of a Crohn’s disease activity index. National Cooperative Crohn’s Disease Study. Gastroenterology. 1976;70:439–44.PubMedGoogle Scholar
  27. 27.
    Ehle H, Horn A. Immunoaffinity chromatography of enzymes. Bioseparation. 1990;1:97–110.PubMedGoogle Scholar
  28. 28.
    Maini RN, Breedveld FC, Kalden JR, Smolen JS, Davis D, Macfarlane JD, et al. Therapeutic efficacy of multiple intravenous infusions of anti-tumor necrosis factor alpha monoclonal antibody combined with low-dose weekly methotrexate in rheumatoid arthritis. Arthritis Rheum. 1998;41:1552–63.PubMedCrossRefGoogle Scholar
  29. 29.
    LoBuglio AF, Wheeler RH, Trang J, Haynes A, Rogers K, Harvey EB, et al. Mouse/human chimeric monoclonal antibody in man: kinetics and immune response. Proc Natl Acad Sci U S A. 1989;86:4220–4.PubMedCrossRefGoogle Scholar
  30. 30.
    Steenholdt C, Bendtzen K, Brynskov J, Thomsen OO, Ainsworth MA. Cut-off levels and diagnostic accuracy of infliximab trough levels and anti-infliximab antibodies in Crohn’s disease. Scand J Gastroenterol. 2011;46:310–8.PubMedCrossRefGoogle Scholar
  31. 31.
    Svenson M, Geborek P, Saxne T, Bendtzen K. Monitoring patients treated with anti-TNF-alpha biopharmaceuticals: assessing serum infliximab and anti-infliximab antibodies. Rheumatology (Oxford). 2007;46:1828–34.CrossRefGoogle Scholar

Copyright information

© Springer 2011

Authors and Affiliations

  • Hirotsugu Imaeda
    • 1
  • Akira Andoh
    • 2
  • Yoshihide Fujiyama
    • 1
  1. 1.Department of MedicineShiga University of Medical ScienceOtsuJapan
  2. 2.Division of Mucosal ImmunologyGraduate School, Shiga University of Medical ScienceOtsuJapan

Personalised recommendations