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Anal Bioanal Chem ; 410(27): 7067-7075, 2018 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-30178083

RESUMEN

IL-6 has an important role in the pathogenesis of autoimmunity and chronic inflammation. Several mAbs that target IL-6 or the IL-6 receptor (IL-6R) have been established and approved for the treatment of various diseases such as multicentric Castleman's disease and rheumatoid arthritis. Quality control of therapeutic antibodies requires accurate determination of bioactivity. However, current cell-based anti-proliferation assays are tedious, time consuming, and result in high variation. We therefore developed a reporter gene assay (RGA) based on an IL-6-dependent DS-1 cell line that stably expressed the reporter luciferase controlled by the serum-induced element (SIE) response element, which was a key element located downstream of the IL-6 signaling pathway. The RGA method demonstrated good performance characteristics after careful optimization, including high specificity, stability, accuracy, precision, and robustness. It also had superior precision and sensitivity. The assay is simple compared with the traditional anti-proliferation assay. This novel RGA based on the IL-6-IL-6R-STAT3 pathway can be useful, in conjunction with the anti-proliferation bioassay, to determine the bioactivity of anti-IL-6/anti-IL-6R therapeutic mAbs. Graphical abstract The mechanism sketch of the reporter gene assay for the bioactivity determination of anti-IL-6/anti-IL-6Rα mAbs.


Asunto(s)
Anticuerpos Monoclonales Humanizados/farmacología , Evaluación Preclínica de Medicamentos , Interleucina-6/antagonistas & inhibidores , Receptores de Interleucina-6/antagonistas & inhibidores , Anticuerpos Monoclonales Humanizados/inmunología , Ingeniería Celular , Línea Celular , Proliferación Celular , Evaluación Preclínica de Medicamentos/métodos , Genes Reporteros , Humanos , Interleucina-6/inmunología , Luciferasas/genética , Luciferasas/inmunología , Receptores de Interleucina-6/inmunología , Proteínas Recombinantes/inmunología
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