Skip to main content

Advertisement

SpringerLink
Log in

Validation of a Drug-Resistant Anti-Adalimumab Antibody Assay to Monitor Immunogenicity in the Presence of High Concentrations of Adalimumab

  • Research Article
  • Published:
The AAPS Journal Aims and scope Submit manuscript

Abstract

With respect to patient safety and long-term efficacy, immunogenicity of therapeutic antibodies remains an important issue. Pre-treatment of samples using either higher temperature or acidification in order to separate drug/anti-drug antibody complexes has been implemented in the traditional bridging assay and an in-house-developed affinity capture elution assay but only a limited drug tolerance was obtained. In this study, we aim to apply a drug-resistant anti-drug antibody assay to adalimumab through a combination of adalimumab/anti-adalimumab antibody complex precipitation and acid dissociation. A linear dose–response curve ranging from 3.1 to 200 ng/mL was obtained in 1/125 diluted serum, allowing detection of anti-adalimumab antibody concentrations up to 25 μg/mL equivalents MA-ADM6A10, a calibrator anti-adalimumab antibody. The cut-off point for detection was determined using 16 samples of adalimumab naïve patients and set at 0.39 μg/mL equivalents. Validation of the assay revealed that no detectable anti-adalimumab antibody concentrations were found in samples with either a positive anti-infliximab antibody concentration, a physiologic concentration of TNFα, or a high concentration of rheumatoid factor. Full recoveries were obtained when various concentrations of adalimumab (0, 1, 10, and 50 μg/mL) were spiked to 1, 2, and 4 μg/mL of MA-ADM6A10. Spiking of 50 μg/mL adalimumab to eight individual sera revealed similar anti-adalimumab antibody concentrations as in the absence of adalimumab, with a Pearson r correlation of 0.99 and an interclass correlation of 0.99. The assay allows accurate evaluation of adalimumab immunogenicity during induction or upon dose intensification and in serum samples not taken at trough.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Abbreviations

ADA:

Anti-drug antibody

ELISA:

Enzyme-linked immunosorbent assay

HRP:

Horse radish peroxidase

IBD:

Inflammatory bowel disease

ICC:

Intra-class correlation coefficient

MA-ADM:

Monoclonal antibody against adalimumab

MA-IFX:

Monoclonal antibody against infliximab

OD:

Optical density

PEG:

Polyethylene glycol

TNFα:

Tumor necrosis factor-alpha

References

  1. West RL, Zelinkova Z, Wolbink GJ, Kuipers EJ, Stokkers PCF, Van Der Woude CJ. Immunogenicity negatively influences the outcome of adalimumab treatment in Crohn’s disease. Aliment Pharmacol Ther. 2008;28(9):1122–6. doi:10.1111/j.1365-2036.2008.03828.x.

    Article  CAS  PubMed  Google Scholar 

  2. Harding FA, Stickler MM, Razo J, DuBridge RB. The immunogenicity of humanized and fully human antibodies. Landes Biosci. 2010;2(3):256–65.

    Google Scholar 

  3. van Schouwenburg PA, Kruithof S, Votsmeier C, van Schie K, Hart MH, de Jong RN, et al. Functional analysis of the anti-adalimumab response using patient-derived monoclonal antibodies. J Biol Chem. 2014;289(50):34482–8. doi:10.1074/jbc.M114.615500.

    Article  PubMed  PubMed Central  Google Scholar 

  4. van Schouwenburg PA, van de Stadt LA, de Jong RN, van Buren EEL, Kruithof S, de Groot E, et al. Adalimumab elicits a restricted anti-idiotypic antibody response in autoimmune patients resulting in functional neutralisation. Ann Rheum Dis. 2012;104–9. doi: 10.1136/annrheumdis-2012-201445.

  5. van Schouwenburg PA, Krieckaert CL, Rispens T, Aarden L, Wolbink GJ, Wouters D. Long-term measurement of anti-adalimumab using pH-shift-anti-idiotype antigen binding test shows predictive value and transient antibody formation. Ann Rheum Dis. 2013;72:1680–6. doi:10.1136/annrheumdis-2012-202407.

    Article  PubMed  Google Scholar 

  6. Shankar G, Shores E, Wagner C, Mire-Sluis A. Scientific and regulatory considerations on the immunogenicity of biologics. Trends Biotechnol. 2006;24(6):274–80. doi:10.1016/j.tibtech.2006.04.001.

    Article  CAS  PubMed  Google Scholar 

  7. Ben-Horin S, Chowers Y. Tailoring anti-TNF therapy in IBD: drug levels and disease activity. Nat Rev Gastroenterol Hepatol. 2014;11(4):243–55. doi:10.1038/nrgastro.2013.253.

    Article  CAS  PubMed  Google Scholar 

  8. Bendtzen K. Immunogenicity of Anti-TNF-α Biotherapies: II. Clinical relevance of methods used for anti-drug antibody detection. Front Immunol. 2015;6(4):1–5. doi:10.3389/fimmu.2015.00109.

    Google Scholar 

  9. Gils A, Vande Casteele N, Poppe R, Van de Wouwer M, Compernolle G, Peeters M, et al. Development of a universal anti-adalimumab antibody standard for interlaboratory harmonization. Ther Drug Monit. 2014;0(0):1–5. doi: 10.1097/FTD.0000000000000074.

  10. van Schouwenburg PA, Kruithof S, Wolbink G, Wouters D, Rispens T. Using monoclonal antibodies as an international standard for the measurement of anti-adalimumab antibodies. J Pharm Biomed Anal. 2016;120:198–201. doi:10.1016/j.jpba.2015.12.040.

    Article  PubMed  Google Scholar 

  11. van Stappen T, Billiet T, Vande Casteele N, Compernolle G, Brouwers E, Vermeire S, et al. An optimized anti-infliximab bridging enzyme-linked immunosorbent assay for harmonization of anti-infliximab antibody titers in patients with inflammatory bowel diseases. Inflamm Bowel Dis. 2015;21(9):2172–7. doi:10.1097/MIB.0000000000000434.

    Article  PubMed  Google Scholar 

  12. van Stappen T, Brouwers E, Vermeire S, Gils A. Validation of a sample pretreatment protocol to convert a drug-sensitive into a drug-tolerant anti-infliximab antibody immunoassay. Drug Test Anal. 2016. doi:10.1002/dta.1968.

    PubMed  Google Scholar 

  13. Bourdage JS, Cook CA, Farrington DL, Chain JS, Konrad RJ. An affinity capture elution (ACE) assay for detection of anti-drug antibody to monoclonal antibody therapeutics in the presence of high levels of drug. J Immunol Methods. 2007;327(1–2):10–7. doi:10.1016/j.jim.2007.07.004.

    Article  CAS  PubMed  Google Scholar 

  14. Zoghbi J, Xu Y, Grabert R, Theobald V, Richards S. A breakthrough novel method to resolve the drug and target interference problem in immunogenicity assays. J Immunol Methods. 2015;426:62–9. doi:10.1016/j.jim.2015.08.002.

    Article  CAS  PubMed  Google Scholar 

  15. Grabert R, Richards S, Theobald V, Xu YX, Zoghbi J. Assays for detecting the presence or amount of an anti-drug antibody. WO 2015123315 A1. 2015, https://www.google.com/patents/WO2015123315A1?cl=en. Accessed 4 November 2016.

  16. Vande Casteele N, Ballet V, Van Assche G, Rutgeerts P, Vermeire S, Gils A. Early serial trough and antidrug antibody level measurements predict clinical outcome of infliximab and adalimumab treatment. Gut. 2012;61:321. doi:10.1136/gut.2010.236869.

    Article  CAS  PubMed  Google Scholar 

  17. Bian S, Stappen TV, Baert F, Compernolle G, Brouwers E, Tops S, et al. Generation and characterization of a unique panel of anti-adalimumab specific antibodies and their application in therapeutic drug monitoring assays. J Pharm Biomed Anal. 2016;125:62–7. doi:10.1016/j.jpba.2016.03.029.

    Article  CAS  PubMed  Google Scholar 

  18. Wang SL, Hauenstein S, Ohrmund L, Shringarpure R, Salbato J, Reddy R, et al. Monitoring of adalimumab and antibodies-to-adalimumab levels in patient serum by the homogeneous mobility shift assay. J Pharm Biomed Anal. 2013;78–79:39–44. doi:10.1016/j.jpba.2013.01.031.

    Article  PubMed  Google Scholar 

  19. van Schouwenburg PA, Bartelds GM, Hart MH, Aarden L, Wolbink GJ, Wouters D. A novel method for the detection of antibodies to adalimumab in the presence of drug reveals “hidden” immunogenicity in rheumatoid arthritis patients. J Immunol Methods. 2010;362(1–2):82–8. doi:10.1016/j.jim.2010.09.005.

    Article  PubMed  Google Scholar 

  20. Kelley M, Ahene AB, Gorovits B, Kamerud J, King LE, McIntosh T, et al. Theoretical considerations and practical approaches to address the effect of anti-drug antibody (ADA) on quantification of biotherapeutics in circulation. AAPS J. 2013;15(3):646–58. doi:10.1208/s12248-013-9468-4.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Dai S, Schantz A, Clements-Egan A, Cannon M, Shankar G. Development of a method that eliminates false-positive results due to nerve growth factor interference in the assessment of fulranumab immunogenicity. AAPS J. 2014;16(3):464–77. doi:10.1208/s12248-014-9581-z.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

This study was financially supported in part grants IOF/KP/12/002 and C22/15/025 from the KU Leuven. SB acknowledges the China Scholarship Council (CSC) for the scholarship. Marc Ferrante is a Senior Clinical Investigator of the Research Foundation—Flanders (FWO), Belgium.

Author information

Authors and Affiliations

  1. Department of Pharmaceutical and Pharmacological Sciences, KU Leuven, Campus Gasthuisberg O&N2, PB 820; Herestraat 49, 3000, Leuven, Belgium

    Sumin Bian & Ann Gils

  2. Department of Gastroenterology and Hepatology, University Hospitals Leuven, KU Leuven, Leuven, Belgium

    Marc Ferrante

Authors
  1. Sumin Bian
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Marc Ferrante
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ann Gils
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ann Gils.

Ethics declarations

Conflict of Interest

MF receives financial support for research from Takeda, lecture fees from Tillotts, Ferring, Boehringer-Ingelheim, Janssen, Chiesi, Falk, Zeria, Mitsubishi Tanabe, MSD, Takeda, and Abbvie and does consultancy for Abbvie, Ferring, MSD, Boehringer-Ingelheim, and Janssen. AG has served as a speaker for MSD, Janssen Biologicals, Pfizer, and Abbvie; as consultant for UCB; and has received Investigator Initiated Research Grants from Pfizer. KU Leuven licensed infliximab and adalimumab ELISA to apDia. SB declares not to have a conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Bian, S., Ferrante, M. & Gils, A. Validation of a Drug-Resistant Anti-Adalimumab Antibody Assay to Monitor Immunogenicity in the Presence of High Concentrations of Adalimumab. AAPS J 19, 468–474 (2017). https://doi.org/10.1208/s12248-016-0018-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1208/s12248-016-0018-8

KEY WORDS

Search

Navigation