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IL-23 is an important pro-inflammatory cytokine of the IL-2 family. Conventional assays for IL-23 activity are based on its ability to proliferate other cell lines, which can be problematic as these cell lines can lose their IL-23 dependence.

This poster describes the development of an IL-23 responsive cell line that can be used for the quantification of IL-23, anti-IL-23 and anti-p40 in as little as 6 hours. Measure IL-23 activity with no response from IL-12 using assay-ready cells available in a convenient 96-well format. Take advantage of the included normalization gene that can be used to compensate for differences in cell numbers and matrix effects.

We have over 10 years’ experience in working with development, validation and production of custom genetically modified cell lines and assay ready cells used in reporter gene assays.

In this poster, we share some of the lessons we learned along the way from this extensive work. In addition, we share our recommendations on what can be considered as an appropriate level of assay read cells characterization using an example from our FGF21 cell line.

Human fibroblast growth factor-21 (FGF21) has the potential to improve glycemic control in patients with type 2 diabetes, as well as the control of weight gain. Analogs of FGF21 are currently being tested in clinical trials in patients with type 2 diabetes and there is a need for a specific assay with enhanced sensitivity both for the quantification of the potency and neutralizing antibody response to FGF21 and related analogues in human serum.

In this poster, we describe the development of a reporter gene cell line that exhibits a high degree of sensitivity to treatment with human FGF21 even in the presence of human serum. These cells can be used in a frozen assay-ready format that confers ease of use for both potency assays and neutralizing antibody assays for monitoring patients treated with FGF21 related products.

In this poster, we demonstrate the robustness of our iLite^® assay system, describing parameters such as accuracy, precision, and plate homogeneity, as well as assay characteristics such as EC50 and LLOQ.

Using an IL-23 cell line as an example, we show that the assay has high repeatability and precision as well as good accuracy. In addition, the assay gives a high fold induction (19x) as well as a lower limit of quantification (LLOQ) in the low nanoliter range (0.1ng/mL). Taken together, the results show that the iLite^® Technology and the Assay Ready format is capable of generating a high performing bioassay, suitable for many applications within the drug continuum.