Laboratory for research and analysis of anti-drug antibodies (ADA) at Karolinska Institutet (KI) and Center for Molecular Medicine (CMM)

Biopharmaceuticals are becoming increasingly important as treatment, especially for patients with severe chronic illnesses. Unfortunately, the immune system reacts against parenterally administered drugs (those given as injections or infusions) and anti-drug antibodies (ADA) might develop. High titers of ADA can block the intended treatment effect and the patient might need to switch treatment to ensure continuous control over the disease. All biopharmaceutical drugs have the potential to activate the immune system, but to what degree depends on the type of drug, formulation, dose, route and frequency of administration, as well as characteristics of the patient and disease. The clinical relevance of ADA depends on the titer, the neutralizing capacity and if they are transient or persistent.

For new drugs and biosimilars the pharma industry needs to provide the regulatory agencies with an evaluation specifying how immunogenic a drug preparation is and what percentage of the patients treated in the clinical trial developed the ADA. These data are unfortunately not directly transferable into clinical practice, which usually consist of a more diverse population of patients subsequently treated over longer periods. In order to guarantee that all patients have the desired effect of their treatment over time, we need to know if their immune system is reacting towards the treatment or not in regular health care practice.

We can measure the ADA and drug levels with various immunological assays and thus at an early stage identify those patients that have lost the effect of the treatment. Our ambition is to, together with the health care system, implement, follow up and review processes whereby we can transfer ADA testing to the health care routine for those patients that are treated with biological drugs and to do research on the potential clinical impact of immunogenicity.

We provide tests for the following biopharmaceuticals:

Neutralizing ADA (nADA or NAb) against interferon beta (IFNbeta) by a bioassay (iLite, Eurodiagnistica, Malmö)

  • Screening for nADA against IFNbeta: 3125 SEK
  • Screening and titration of nADA against IFNbeta (for the around 20% expected to become positive): 5000 SEK

ADA against natalizumab (Tysabri) by ELISA (BiogenIdec, ”bridging” ELISA)

  • Screening of ADA against Tysabri: 2500 SEK

Analysis of ADA against rituximab (Mabthera) by electrochemiluminiscens (ECL) on the Meso Scale Discovery (MSD) platform

  • Screening, including titration of positive samples: 4200 SEK

From June 2021 we are official sub-contractors to Karolinska University Hospital, Clinical Immunology and transfusion medicine, the routine laboratory whom also provides other ADA tests and from then ADA against rituximab can be ordered through TakeCAre.

From November 2007 the test results of the ADA analyses for MS patients are registered in the Swedish MS registry. You can register MS patients and download referral forms from this home page, which is part of the Swedish Neuroregister:

Swedish Neuroregister

Referral forms can also be downloaded directly from here:

Interferon beta nADA referral form

Tysabri (natalizumab) referral form

Mabthera (rituximab) referral form

For other patient groups than MS, the test results will be sent by paper to the address of the clinic noted on the referral form.

For patients treated with TNF blocking drugs, we recommend to start with screening of drug level in trough by ELISA that can be ordered through the TakeCare system and which is done by Clinical Immunology at the Karolinska University Hospital. Samples with very low trough level of drug (<0.2 µg/ml) will be tested for ADA by ELISA at Clinical Immunology at the Karolinska University Hospital. Samples with low drug level but above 0.2 µg/ml might have a suboptimal effect due to immune complexes formed between the drug and the ADA. These can be detected by the PandA method using ECL on MSD at the ADA lab. KI/CMM and ordered through this referral form:

PandA infliximab referral form

You are welcome to send in your samples!

Associate professor Anna Fogdell-Hahn, Dr. Katharina Fink (MD/PhD) and professor Jan Hillert


Rituximab (Mabthera and MS)

Rituximab in multiple sclerosis: Frequency and clinical relevance of anti-drug antibodies.
Dunn N, Juto A, Ryner M, Manouchehrinia A, Piccoli L, Fink K, et al
Mult Scler 2018 08;24(9):1224-1233

IFNbeta ADA and MS

Development and validation of cell-based luciferase reporter gene assays for measuring neutralizing anti-drug antibodies against interferon beta.
Hermanrud C, Ryner M, Luft T, Jensen PE, Ingenhoven K, Rat D, et al
J Immunol Methods 2016 Mar;430():1-9

Development and Validation of an Enzyme-Linked Immunosorbent Assay for the Detection of Binding Anti-Drug Antibodies against Interferon Beta.
Ingenhoven K, Kramer D, Jensen PE, Hermanrud C, Ryner M, Deisenhammer F, et al
Front Neurol 2017 ;8():305

Clinical practice of analysis of anti-drug antibodies against interferon beta and natalizumab in multiple sclerosis patients in Europe: A descriptive study of test results.
Link J, Ramanujam R, Auer M, Ryner M, Hässler S, Bachelet D, et al
PLoS One 2017 ;12(2):e0170395

Occurrence of Anti-Drug Antibodies against Interferon-Beta and Natalizumab in Multiple Sclerosis: A Collaborative Cohort Analysis.
Bachelet D, Hässler S, Mbogning C, Link J, Ryner M, Ramanujam R, et al
PLoS One 2016 ;11(11):e0162752

Anti-interferon beta antibody titers strongly correlate between two bioassays and in vivo biomarker expression, and indicates that a titer of 150 TRU/mL is a biologically functional cut-point.
Hermanrud C, Ryner ML, Engdahl E, Fogdell-Hahn A
J Interferon Cytokine Res 2014 Jul;34(7):498-504

Prevalence of anti-drug antibodies against interferon beta has decreased since routine analysis of neutralizing antibodies became clinical practice.
Jungedal R, Lundkvist M, Engdahl E, Ramanujam R, Westerlind H, Sominanda A, et al
Mult Scler 2012 Dec;18(12):1775-81

Inhibition of endogenous interferon beta by neutralizing antibodies against recombinant interferon beta.
Sominanda A, Lundkvist M, Fogdell-Hahn A, Hemmer B, Hartung HP, Hillert J, et al
Arch Neurol 2010 Sep;67(9):1095-101

Multiple sclerosis patients lacking oligoclonal bands in the cerebrospinal fluid are less likely to develop neutralizing antibodies against interferon beta.

Lundkvist M, Greiner E, Hillert J, Fogdell-Hahn A
Mult Scler 2010 Jul;16(7):796-800

Neutralizing antibodies against interferon β: fluctuation is modest and titre dependent.
Sominanda A, Hillert J, Fogdell-Hahn A
Eur J Neurol 2009 Jan;16(1):21-6

Comparative study of four different assays for the detection of binding antibodies against interferon-beta.
Gneiss C, Brugger M, Millonig A, Fogdell-Hahn A, Rudzki D, Hillert J, et al
Mult Scler 2008 Jul;14(6):830-6

In vivo bioactivity of interferon-beta in multiple sclerosis patients with neutralising antibodies is titre-dependent.
Sominanda A, Hillert J, Fogdell-Hahn A
J Neurol Neurosurg Psychiatry 2008 Jan;79(1):57-62

Interferon beta preparations for the treatment of multiple sclerosis patients differ in neutralizing antibody seroprevalence and immunogenicity.
Sominanda A, Rot U, Suoniemi M, Deisenhammer F, Hillert J, Fogdell-Hahn A
Mult Scler 2007 Mar;13(2):208-14

Natalizumab (Tysabri) ADA and MS

Characterization of anti-natalizumab antibodies in multiple sclerosis patients.
Lundkvist M, Engdahl E, Holmén C, Movérare R, Olsson T, Hillert J, et al
Mult Scler 2013 May;19(6):757-64

Fatal neuroinflammation in a case of multiple sclerosis with anti-natalizumab antibodies.
Svenningsson A, Dring AM, Fogdell-Hahn A, Jones I, Engdahl E, Lundkvist M, et al
Neurology 2013 Mar;80(10):965-7

Occurrence of antibodies against natalizumab in relapsing multiple sclerosis patients treated with natalizumab.

Sørensen PS, Jensen PE, Haghikia A, Lundkvist M, Vedeler C, Sellebjerg F, et al
Mult Scler 2011 Sep;17(9):1074-8

A Swedish national post-marketing surveillance study of natalizumab treatment in multiple sclerosis.

Holmén C, Piehl F, Hillert J, Fogdell-Hahn A, Lundkvist M, Karlberg E, et al
Mult Scler 2011 Jun;17(6):708-19

Review articles:

Antidrug Antibodies: B Cell Immunity Against Therapy.
Fogdell-Hahn A
Scand J Immunol 2015 Sep;82(3):184-90

Standardizing terms, definitions and concepts for describing and interpreting unwanted immunogenicity of biopharmaceuticals: recommendations of the Innovative Medicines Initiative ABIRISK consortium.
Rup B, Pallardy M, Sikkema D, Albert T, Allez M, Broet P, et al
Clin Exp Immunol 2015 Sep;181(3):385-400

The case for measuring anti-drug antibodies in people with multiple sclerosis.

Lundkvist Ryner M, Farrell RA, Fogdell-Hahn A
Expert Rev Clin Immunol 2014 Jun;10(6):697-9

Anti-drug antibodies.
Warnke C, Hermanrud C, Lundkvist M, Fogdell-Hahn A.
Drugs and Therapy Studies 2012; volume 2:e11