Reporter cell assay-based functional quantification of TNF-α-antagonists in serum

• Verfasst von: Schuster, Lukas [VerfasserIn]
• Verfasst von: Sauter, Max [VerfasserIn]
• Verfasst von: Uhl, Philipp [VerfasserIn]
• Verfasst von: Meid, Andreas [VerfasserIn]
• Verfasst von: Haefeli, Walter E. [VerfasserIn]
• Verfasst von: Weiß, Johanna [VerfasserIn]
• Verfasst von: Theile, Dirk [VerfasserIn]
• Titel: Reporter cell assay-based functional quantification of TNF-α-antagonists in serum
• Titelzusatz: a proof-of-principle study for adalimumab
• Verf.angabe: Lukas Schuster, Max Sauter, Philipp Uhl, Andreas Meid, Walter E. Haefeli, Johanna Weiss, Dirk Theile
• E-Jahr: 2020
• Jahr: 26 February 2020
• Umfang: 5 S.
• Fussnoten: Im Text wird alpha als griechisches Zeichen dargestellt ; Gesehen am 24.04.2020
• Titel Quelle: Enthalten in: Analytical biochemistry
• Ort Quelle: San Diego, Calif. : Elsevier, 1960
• Jahr Quelle: 2020
• Band/Heft Quelle: 596(2020) Arikel-Nummer 113646, 5 Seiten
• ISSN Quelle: 1096-0309
• DOI: doi:10.1016/j.ab.2020.113646
• Datenträger: Online-Ressource
• Sprache: eng
• Sach-SW: Adalimumab
• Sach-SW: HEK-Blue™-hTLR3
• Sach-SW: NF-κB
• Sach-SW: Reporter cell assay
• Sach-SW: Tumor necrosis factor
• K10plus-PPN: 169607617X

Abstract

Quantification of therapeutic antibodies is commonly based on physico-chemical assays such as enzyme-linked immunoabsorption assays (ELISA) and lately on mass spectrometry. However, the functional integrity of evaluated immunoglobulins is yet not assessed. Consequently, a commercially available reporter cell line was used to quantify the functional concentration of the anti-tumor necrosis factor alpha (TNF-α) antibody adalimumab present in serum of a healthy beagle dog treated with 3 mg intravenous adalimumab (Humira®). HEK-Blue™-hTLR3 cells express a secreted alkaline phosphatase under the control of a nuclear factor kappa B (NF-κB) response element. Its enzymatic activity can be recorded using colorimetry, which reports activity of extracellular NF-κB stimuli such as TNF-α. Using an adalimumab concentration-response calibration curve, the functional concentration of serum adalimumab was estimated to be 4.9 ± 1.4 μg/ml, which was in excellent agreement with ELISA results (4.8 μg/ml). The obtained data suggest that this simple, easy-to-handle reporter cell assay can be used for the functional quantification of adalimumab present in samples from in vitro or pre-clinical in vivo experiments. Moreover, this assay could be used in vitro to compare the pharmacodynamics of adalimumab biosimilars or different anti-TNF-α compounds, respectively.