Article Text

AB1297 Bridging ELISA as a secreening assay to monitor immunogenicity in routine clinical practice
  1. S. Garcês1,2,
  2. J. Demengeot2,
  3. J. Canas-da-Silva1,
  4. L. Aarden3
  1. 1Rheumatology, Hospital Garcia De Orta, Almada
  2. 2Immunology, Instituto Gulbenkian Ciência, Oeiras, Portugal
  3. 3Immunology, Sanquin, Amsterdam, Netherlands


Background The clinical and scientific relevance of monitoring immunogenicity in clinical practice have been recognized and recommended by the European Agency of Medicine (EMA)1. However, its assessment is technically challenging and no consensus exists about the way immunogenicity can be monitored. Newly developed fluid-phase radio-immuno assays (RIA) have proven quite effective in detecting anti-drug antibodies and as been presented as a “gold standard” method to assess immunogenicity2,3. However, RIA requires high doses of radioactivity and special conditions, preventing its use as a routine assay. By contrast, enzyme-linked immunoassay (ELISA) it’s a simple and cheap method, ideal candidate to be used as a high-throughput screening assay. Bridging ELISA is a particular type of ELISA with increased specificity over the conventional ELISAs methods. Several optimizations of this method were made so that it can become a good screeningassay to monitor drug immunogenicity in routine clinical practice.

Objectives We tested, in a same cohort of patients receiving infliximab, this newly optimized bridging ELISA in comparison with a fluid-phase antigen-binding RIA to quantify anti-infliximab antibodies.

Methods A total of 82 consecutive patients were evaluated (38 rheumatoid arthritis patients, 27 ankylosing spondylitis, 9 psoriatic arthritis and 18 patients with inflammatory bowel disease), 61 females, with a mean age of 41 (4.2) years, that were receiving Infliximab in a dosage of 3-5mg/kg every 6 or 8 weeks for a mean period of 3.5 (2.0) years. Blood samples were collected immediately before the next infliximab infusion. Anti-infliximab antibodies were quantified by 1) Bridging ELISA where the antibodies bind to the infliximab coated in a solid phase and revealed by the addition of biotinylated infliximab and by 2) fluid-phase RIA-ABA that uses a sepharose-immobilized protein A, IgG total and IgG4-specific, to bind IgGs in the patient’s serum. Anti-infliximab specific IgGs are revealed by theaddition of 125I-labeled infliximab F(ab’)2. A simple ELISA method was used to quantify serum infliximab levels,based on the principle that infliximab are captured through their ability to bind immobilized TNFα on a solid phase. The binding assessment is revealed by incubation with biotinylated rabbit IgG directed to infliximab idiotype. Therapeutic response was assessed according to validated criteria established for each disease.

Results A total of 22 (27%) were tested positive for the presence of anti-infliximab abs using RIA, coinciding with the samples that were also positives in Bridging ELISA. Bridging ELISA cannot detect monovalent IgG4. No samples testing exclusively IgG4 specific anti-infliximab were detected. All patients (100%) with detectable anti-infliximab antibodies had undetectable serum trough drug levels and were not able to sustain the therapeutic response.

Conclusions By its simplicity, cost and robustness, Bridging ELISA is a suitable test to be implemented in routine clinical practice as a screening assay to monitor drug immunogenicity.

  1. E.M.A.

  2. L et al. Curr Opin Immunol, 2008; 20: 431-5.

  3. Svenson M et al. Rheumatology (Oxford), 2007; 46: 1828-34.

Disclosure of Interest None Declared

Statistics from

Request permissions

If you wish to reuse any or all of this article please use the link below which will take you to the Copyright Clearance Center’s RightsLink service. You will be able to get a quick price and instant permission to reuse the content in many different ways.