| Autor: |
Federica R, Achini-Gutzwiller, Cornelia M, Jol-van der Zijde, Anja M, Jansen-Hoogendijk, Arjan C, Lankester, Robbert G M, Bredius, Maarten J D, van Tol, Dirk Jan A R, Moes, Marco W, Schilham |
| Rok vydání: |
2022 |
| Zdroj: |
Therapeutic drug monitoring. |
| ISSN: |
1536-3694 |
| Popis: |
Alemtuzumab is a humanized monoclonal antibody that targets the CD52 glycoprotein expressed on most lymphocytes, subsequently inducing complement- and antibody-mediated cytotoxicity. Owing to its ability to induce profound immune depletion, alemtuzumab is frequently used in patients prior to allogeneic hematopoietic stem cell transplantation to prevent graft rejection and acute graft-versus-host disease. In this clinical context, a stable immunoassay with high sensitivity and specificity to determine alemtuzumab levels is essential for performing pharmacokinetic and pharmacodynamic analyses; however, the available methods have several limitations. Here, we report the successful development and validation of an efficient and highly sensitive enzyme-linked immunosorbent assay (ELISA) technique based on commercially available reagents to quantify alemtuzumab in human serum or plasma.This ELISA technique was developed and validated in accordance with the European Medicines Agency guidelines on bioanalytical method validation.The assay sensitivity (lower limit of quantification) is 0.5 ng mL-1 and the dynamic range is 0.78-25 ng mL-1. To accommodate quantification of peak concentration as well as concentrations below the lympholytic level (0.1 µg mL-1), patients' serum samples were prediluted 20-400 times according to the expected alemtuzumab concentration. The overall within-run accuracy was between 96 and 105%, whereas overall within-run precision (coefficient of variation, CV) was between 3 and 9%. The between-runs assessment provided an overall accuracy between 86 and 95% and an overall CV between 5 and 14%.The developed assay provides accurate insight into alemtuzumab exposure and its effects on the clinical response to treatment, which is key to optimizing treatment strategies. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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