Development and validation of cell-based luciferase reporter gene assays for measuring neutralizing anti-drug antibodies against interferon beta

Hermanrud, Christina; Ryner, Malin; Luft, Thomas; Jensen, Poul Erik; Ingenhoven, Kathleen; Rat, Dorothea; Deisenhammer, Florian; Sørensen, Per Soelberg; Pallardy, Marc; Sikkema, Daniel J.; Bertotti, Elisa; Kramer, Daniel; Creeke, Paul I.; Fogdell‐Hahn, Anna

doi:10.1016/j.jim.2016.01.004

Cited by 12 publications

(11 citation statements)

References 212 publications

(239 reference statements)

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“…Nevertheless, the difference in sensitivity between ADA assay methods highlights the importance of using the same validated assay across laboratories to get comparable results when performing ADA testing. One of the first tasks of the ABIRISK project was to establish and validate assays for binding (manuscript submitted) and neutralizing ADA against IFNβ [ 31 ] and ADA against natalizumab (manuscript in progress), since methodological variations among different assays can impact the results reported between studies. The clinical relevance of low titer ADA can be questioned and therefore the most sensitive assays may not be required for routine monitoring of ADA in the clinic.…”

Section: Discussionmentioning

confidence: 99%

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

et al. 2017

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Antibodies against biopharmaceuticals (anti-drug antibodies, ADA) have been a well-integrated part of the clinical care of multiple sclerosis (MS) in several European countries. ADA data generated in Europe during the more than 10 years of ADA monitoring in MS patients treated with interferon beta (IFNβ) and natalizumab have been pooled and characterized through collaboration within a European consortium. The aim of this study was to report on the clinical practice of ADA testing in Europe, considering the number of ADA tests performed and type of ADA assays used, and to determine the frequency of ADA testing against the different drug preparations in different countries. A common database platform (tranSMART) for querying, analyzing and storing retrospective data of MS cohorts was set up to harmonize the data and compare results of ADA tests between different countries. Retrospective data from six countries (Sweden, Austria, Spain, Switzerland, Germany and Denmark) on 20,695 patients and on 42,555 samples were loaded into tranSMART including data points of age, gender, treatment, samples, and ADA results. The previously observed immunogenic difference among the four IFNβ preparations was confirmed in this large dataset. Decreased usage of the more immunogenic preparations IFNβ-1a subcutaneous (s.c.) and IFNβ-1b s.c. in favor of the least immunogenic preparation IFNβ-1a intramuscular (i.m.) was observed. The median time from treatment start to first ADA test correlated with time to first positive test. Shorter times were observed for IFNβ-1b-Extavia s.c. (0.99 and 0.94 years) and natalizumab (0.25 and 0.23 years), which were introduced on the market when ADA testing was already available, as compared to IFNβ-1a i.m. (1.41 and 2.27 years), IFNβ-1b-Betaferon s.c. (2.51 and 1.96 years) and IFNβ-1a s.c. (2.11 and 2.09 years) which were available years before routine testing began. A higher rate of anti-IFNβ ADA was observed in test samples taken from older patients. Testing for ADA varies between different European countries and is highly dependent on the policy within each country. For drugs where routine monitoring of ADA is not in place, there is a risk that some patients remain on treatment for several years despite ADA positivity. For drugs where a strategy of ADA testing is introduced with the release of the drug, there is a reduced risk of having ADA positive patients and thus of less efficient treatment. This indicates that potential savings in health cost might be achieved by routine analysis of ADA.

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Section: Discussionmentioning

confidence: 99%

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

et al. 2017

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“…From 2757 MS patients recruited in Sweden and Germany and treated with three different IFNβ preparations (Table 1 ), bADA levels were measured by capture ELISA [ 20 ] and nADA titers using a luciferase-based bioassay [ 21 , 22 ] (Additional files 1 - 3 ). The bADA levels were correlated with nADA presence (Spearman ρ = 0.66) and nADA titers ( ρ = 0.71).…”

Section: Resultsmentioning

confidence: 99%

“…Binding ADA levels were measured by capture ELISA [ 20 ] at a single site (Munich) and were calculated from optical densities using a standard curve (Additional file 2 ). For the assessment of nADA titers, measured as the inverse of serum dilutions using a luciferase-based bioassay [ 21 ], samples were first screened, and titration was only conducted for samples positive during screening [ 22 ]. Assessment of nADA titers was conducted at two separate sites (Innsbruck and Copenhagen), to which samples were assigned using adaptive randomization to minimize differences regarding the recruitment site, gender, the age at the blood draw, the IFNβ treatment preparation, and the total duration of IFNβ treatment.…”

Section: Methodsmentioning

confidence: 99%

Treatment- and population-specific genetic risk factors for anti-drug antibodies against interferon-beta: a GWAS

Andlauer

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Martin

et al. 2020

BMC Med

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Background Upon treatment with biopharmaceuticals, the immune system may produce anti-drug antibodies (ADA) that inhibit the therapy. Up to 40% of multiple sclerosis patients treated with interferon β (IFNβ) develop ADA, for which a genetic predisposition exists. Here, we present a genome-wide association study on ADA and predict the occurrence of antibodies in multiple sclerosis patients treated with different interferon β preparations. Methods We analyzed a large sample of 2757 genotyped and imputed patients from two cohorts (Sweden and Germany), split between a discovery and a replication dataset. Binding ADA (bADA) levels were measured by capture-ELISA, neutralizing ADA (nADA) titers using a bioassay. Genome-wide association analyses were conducted stratified by cohort and treatment preparation, followed by fixed-effects meta-analysis. Results Binding ADA levels and nADA titers were correlated and showed a significant heritability (47% and 50%, respectively). The risk factors differed strongly by treatment preparation: The top-associated and replicated variants for nADA presence were the HLA-associated variants rs77278603 in IFNβ-1a s.c.- (odds ratio (OR) = 3.55 (95% confidence interval = 2.81–4.48), p = 2.1 × 10−26) and rs28366299 in IFNβ-1b s.c.-treated patients (OR = 3.56 (2.69–4.72), p = 6.6 × 10−19). The rs77278603-correlated HLA haplotype DR15-DQ6 conferred risk specifically for IFNβ-1a s.c. (OR = 2.88 (2.29–3.61), p = 7.4 × 10−20) while DR3-DQ2 was protective (OR = 0.37 (0.27–0.52), p = 3.7 × 10−09). The haplotype DR4-DQ3 was the major risk haplotype for IFNβ-1b s.c. (OR = 7.35 (4.33–12.47), p = 1.5 × 10−13). These haplotypes exhibit large population-specific frequency differences. The best prediction models were achieved for ADA in IFNβ-1a s.c.-treated patients. Here, the prediction in the Swedish cohort showed AUC = 0.91 (0.85–0.95), sensitivity = 0.78, and specificity = 0.90; patients with the top 30% of genetic risk had, compared to patients in the bottom 30%, an OR = 73.9 (11.8–463.6, p = 4.4 × 10−6) of developing nADA. In the German cohort, the AUC of the same model was 0.83 (0.71–0.92), sensitivity = 0.80, specificity = 0.76, with an OR = 13.8 (3.0–63.3, p = 7.5 × 10−4). Conclusions We identified several HLA-associated genetic risk factors for ADA against interferon β, which were specific for treatment preparations and population backgrounds. Genetic prediction models could robustly identify patients at risk for developing ADA and might be used for personalized therapy recommendations and stratified ADA screening in clinical practice. These analyses serve as a roadmap for genetic characterizations of ADA against other biopharmaceutical compounds.

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“…Within the ABIRISK consortium, this assay will be used for testing for the development of anti-IFN-β antibodies in patient samples prospectively collected after start of therapy with IFN-β and compared with the results from bioassays used for the detection of antibodies with neutralizing capacity (8). Furthermore, newly developed and unique monoclonal anti-IFN-β antibodies of human origin will be tested, and the assay sensitivity and drug tolerance reassessed for this PC.…”

Section: Resultsmentioning

confidence: 99%

“…The aim of this study was to develop and validate a method for the detection of ADA toward IFN-β in human serum, following the published ABIRISK recommendations. These recommendations include a typical three-tiered approach, in which the samples are first screened and confirmed for binding ADA in a two-step analysis as described in this manuscript, and then further analyzed for neutralizing capacity and titers in a bio-assay recently published (8). The assay validation set forth in this article followed established industry and regulatory guidelines (9–13).…”

Section: Introductionmentioning

confidence: 99%

Development and Validation of an Enzyme-Linked Immunosorbent Assay for the Detection of Binding Anti-Drug Antibodies against Interferon Beta

et al. 2017

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objective: To develop and validate a method for the detection of binding anti-drug antibodies (ADAs) against interferon beta (IFN-β) in human serum as part of a European initiative (ABIRISK) aimed at the prediction and analysis of clinical relevance of antibiopharmaceutical immunization to minimize the risk.Method: A two-tiered bridging enzyme-linked immunosorbent assay (ELISA) format was selected and validated according to current recommendations. Screening assay: ADA in serum samples form complexes with immobilized IFN-β and biotinylated IFN-β, which are then detected using HRP labeled Streptavidin and TMB substrate. Confirmation assay: Screen "putative positive" samples are tested in the presence of excess drug (preincubation of sera with 0.3 µg/mL of soluble IFN-β) and percentage of inhibition is calculated.

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Development and validation of cell-based luciferase reporter gene assays for measuring neutralizing anti-drug antibodies against interferon beta

Cited by 12 publications

References 212 publications

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

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

Treatment- and population-specific genetic risk factors for anti-drug antibodies against interferon-beta: a GWAS

Development and Validation of an Enzyme-Linked Immunosorbent Assay for the Detection of Binding Anti-Drug Antibodies against Interferon Beta

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