Development and Validation of an Enzyme-Linked Immunosorbent Assay to Measure Adalimumab Concentration

Desvignes, Céline; Edupuganti, Soujanya Ratna; Darrouzain, François; Duveau, Anne-Claire; Loercher, Amy; Paintaud, Gilles; Mulleman, Denis

doi:10.4155/bio.15.30

Cited by 16 publications

(12 citation statements)

References 17 publications

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“…Adalimumab concentration was measured by using a validated ELISA. The limit of detection was 0.022 µg/mL; the concentrations for the lower and upper limits of quantification were 0.073 and 9 µg/mL, respectively 13. All biological analyses were performed after study completion without the knowledge of clinical data or group of randomisation.…”

Section: Methodsmentioning

confidence: 99%

Methotrexate effect on immunogenicity and long-term maintenance of adalimumab in axial spondyloarthritis: a multicentric randomised trial

et al. 2020

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ObjectivesAnti-drug antibodies (ADA) are responsible for decreased adalimumab efficacy in axial spondyloarthritis (SpA). We aimed to evaluate the ability of methotrexate (MTX) to decrease adalimumab immunisation.MethodsA total of 110 patients eligible to receive adalimumab 40 mg subcutaneously (s.c.) every other week were randomised (1:1 ratio) to receive, 2 weeks before adalimumab (W-2) and weekly, MTX 10 mg s.c. (MTX+) or not (MTX−). ADA detection and adalimumab serum concentration were assessed at weeks 4 (W4), 8 (W8), 12 (W12) and 26 (W26) after starting adalimumab (W0). The primary outcome was the proportion of patients with ADA at W26. Four years after the study completion, we retrospectively analysed adalimumab maintenance in relation with MTX co-treatment duration.ResultsWe analysed data for 107 patients (MTX+; n=52; MTX-; n=55). ADA were detected at W26 in 39/107 (36.4%) patients: 13/52 (25%) in the MTX+ group and 26/55 (47.3%) in the MTX− group (p=0.03). Adalimumab concentration was significantly higher in the MTX+ than MTX− group at W4, W8, W12 and W26. The two groups did not differ in adverse events or efficacy. In the follow-up study, MTX co-treatment >W26 versus no MTX or ≤W26 was significantly associated with adalimumab long-term maintenance (p=0.04).ConclusionMTX reduces the immunogenicity and ameliorate the pharmacokinetics of adalimumab in axial SpA. A prolonged co-treatment of MTX>W26 seems to increase adalimumab long-term maintenance.

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Section: Methodsmentioning

confidence: 99%

Methotrexate effect on immunogenicity and long-term maintenance of adalimumab in axial spondyloarthritis: a multicentric randomised trial

et al. 2020

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“…A majority of the ELISAs were commercial kits (n=10) but in-house assays were also performed (n=2). ELISA formats varied (53)(54)(55). In some cases, other anti-TNF-a therapeutics could also be detected, however, most were specific for adalimumab.…”

Section: Study Design and Assays For Assessing Adalimumab Levelsmentioning

confidence: 99%

The First WHO International Standard for Adalimumab: Dual Role in Bioactivity and Therapeutic Drug Monitoring

et al. 2021

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The expanded availability of adalimumab products continues to widen patient access and reduce costs with substantial benefit to healthcare systems. However, the long-term success of these medicines is highly dependent on maintaining consistency in quality, safety and efficacy while minimizing any risk of divergence during life-cycle management. In recognition of this need and demand from global manufacturers, the World Health Organization (WHO) Expert Committee on Biological standardization established the WHO 1st International standard (IS) for Adalimumab (coded 17/236) in October 2019 with a defined unitage ascribed to each of the individual bioactivities evaluated in the study namely, TNF-α binding, TNF-α neutralization, complement dependent cytotoxicity and antibody-dependent cellular cytotoxicity. For development of the IS, two candidate standards were manufactured as per WHO recommendations. Analysis of extensive datasets generated by testing of a common set of samples including the candidate standards by multiple stakeholders including regulatory agencies using their own qualified assays in a large international collaborative study showed comparable biological activity for the tested candidates for the different activities. Use of a common standard significantly decreased the variability of bioassays and improved agreement in potency estimates. Data from this study clearly supports the utility of the IS as an important tool for assuring analytical assay performance, for bioassay calibration and validation, for identifying and controlling changes in bioactivity during life-cycle management and for global harmonization of adalimumab products. In addition, in a separate multi-center study which included involvement of hospital and clinical diagnostic laboratories, the suitability of the adalimumab IS for therapeutic drug monitoring assays was examined by analysis of data from testing of a common blind coded panel of adalimumab spiked serum samples representative of the clinical scenario along with the IS and in-house standards in diverse immunoassays/platforms. Both commercially available and in-house assays that are routinely used for assessing adalimumab trough levels were included. Excellent agreement in estimates for adalimumab content in the spiked samples was observed regardless of the standard or the method with inter-laboratory variability also similar regardless of the standard employed. This data, for the first time, provides support for the extended applicability of the IS in assays in use for therapeutic drug monitoring based on the mass content of the IS. The adalimumab IS, in fulfilling clinical demand, can help toward standardizing and harmonizing clinical monitoring assays for informed clinical decisions and/or personalized treatment strategies for better patient outcomes. Collectively, a significant role for the adalimumab IS in assuring the quality, safety and efficacy of adalimumab products globally is envisaged.

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“…Assay validation followed the procedure previously described [18][19][20]. Notably, coating was performed overnight at +4°C, by incubating 100 µL/well of 2 mg/l sVEGFR-2 in carbonatebicarbonate buffer.…”

Section: Assay Proceduresmentioning

confidence: 99%

“…The precision and accuracy were assessed as previously described [18][19][20]. Briefly, intra-and inter-day precisions and accuracies were defined as coefficient of variation (% CV = Standard deviation (SD)/mean) and absolute bias (% RE, where relative error (RE) = [measured valueactual value]/actual value), respectively.…”

Section: Precision Accuracy Limit Of Quantificationmentioning

confidence: 99%

A Robust Enzyme-Linked Immunosorbent Assay to Measure Serum Ramucirumab Concentrations

et al. 2021

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Aim: Ramucirumab, an anti-VEGFR2 monoclonal antibody, has been approved for the treatment of metastatic gastric and colorectal cancer. An assay measuring ramucirumab serum concentrations was needed to investigate its pharmacokinetics and concentration–response relationship. Results: An ELISA was developed and validated according to the international guidelines for ligand-binding assays. Ramucirumab calibration standards ranged from 0.125 to 40 mg/l. Low, middle and high quality controls were spiked at 0.2, 4 and 8 mg/l, respectively. The limits of quantification were established to be 0.125 and 10 mg/l for LLOQ and ULOQ, respectively. No cross-reactivity with anti-VEGF or anti-EGFR was detected. Conclusion: This in-house-developed ELISA is sensitive, accurate, reproducible and suitable for pharmacokinetic studies of ramucirumab.

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Development and Validation of an Enzyme-Linked Immunosorbent Assay to Measure Adalimumab Concentration

Abstract: This method is accurate, reproducible and may be useful for PK studies and for therapeutic drug monitoring of adalimumab.

Cited by 16 publications

References 17 publications

Methotrexate effect on immunogenicity and long-term maintenance of adalimumab in axial spondyloarthritis: a multicentric randomised trial

Methotrexate effect on immunogenicity and long-term maintenance of adalimumab in axial spondyloarthritis: a multicentric randomised trial

The First WHO International Standard for Adalimumab: Dual Role in Bioactivity and Therapeutic Drug Monitoring

A Robust Enzyme-Linked Immunosorbent Assay to Measure Serum Ramucirumab Concentrations

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