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Issue 69, 2019
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A simple and robust reporter gene assay for measuring the bioactivity of anti-RANKL therapeutic antibodies

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Abstract

RANKL (receptor activator of nuclear factor κB ligand) plays a key role in the differentiation, activation and survival of osteoclasts. Denosumab, which targets RANKL, is approved for osteoporosis or bone loss that has a high risk for fracture and bone metastases from solid tumors. Bioactivity determination is essential for the safety and efficacy of therapeutic antibodies. At present, the mechanism of action (MOA) based bioassay for anti-RANKL monoclonal antibodies (mAbs) is the measurement of tartrate resistant acid phosphatase (TRAP) activity, which takes about five days and has complex operation and relatively high variation. In this study, we developed a reporter gene assay (RGA) based on a RAW264.7 cell line stably expressing luciferase reporter under the control of nuclear factor-κB (NF-κB) response elements. After optimizing the key parameters, the validation results based on ICH-Q2 not only show superior specificity, precision, linearity, accuracy and passage stability, but also a short duration and simple operation. These results demonstrate the RGA based on the RANKL–RANK–NF-κB pathway can be an excellent alternative for measuring the bioactivity of anti-RANKL mAbs.

Graphical abstract: A simple and robust reporter gene assay for measuring the bioactivity of anti-RANKL therapeutic antibodies

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Article information


Submitted
12 Sep 2019
Accepted
16 Nov 2019
First published
03 Dec 2019

This article is Open Access

RSC Adv., 2019,9, 40196-40202
Article type
Paper

A simple and robust reporter gene assay for measuring the bioactivity of anti-RANKL therapeutic antibodies

C. Yu, L. Wang, Y. Ni and J. Wang, RSC Adv., 2019, 9, 40196
DOI: 10.1039/C9RA07328K

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    [Original citation] - Published by The Royal Society of Chemistry.

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