Find out how the iLite® technology provides a single seamless solution for accelerating drug development

iLite® Technology

A versatile tool for the whole drug development process

The iLite technology, based on a reporter gene assay format, provides a seamless solution for applications extending across the whole drug development process, as well as for monitoring of biological drugs.

The technology can be developed for virtually any pharmaceutical target and allows an easy, rapid and accurate test format for measurement and quantification of drug activity and immunogenicity. The cells are supplied in an Assay Ready Format, removing the need for cell-culturing and allowing a seamless transfer of the assay between departments or to third party collaborators.

By combining unique features such as a normalization readout and chimeric transcription factors with a highly flexible product format, the iLite technology helps you make the most of your bioassay.

The dual reporter gene system - How it works

When a ligand binds to its receptor, a specific intracellular signaling pathway is activated, triggering the transcription of a specific reporter gene construct coding for luciferase. When a substrate is added, the luciferase generates light, and the amount of luciferase and thereby the amount and activity of the ligand can then be measured as light emission using a luminometer.

A unique feature of the iLite cells is that they also contain a second reporter gene which is under the control of a constitutive promoter, meaning that it is expressed continuously in any living cell. In other words, the amount of light generated from this second reporter gene will be correlated to the number of living cells, and can therefore be used for normalizing results to compensate for differences in cell number.

Normalization Gene

The second reporter gene works as a kind of internal control, and will compensate for differences in cell number, obviating the need for counting the number of cells applied in each well. Another major and unique benefit is the possibility of compensating for serum matrix effects, or other complex matrices that might have a toxic effect on the cells. Normalization can also be used if the luciferase is sensitive to the compound you’re analyzing, as can occur during high-throughput screening procedures.

Assay Ready Format

iLite cells are delivered as Assay Ready Cells, and stored at -80°C. The Assay Ready format removes the need for cell culture and continuous maintenance of cells. The cells are simply thawed and diluted before use in the assay. Besides significant reductions in the amount of labor required and assay turnaround times, the Assay Ready Cell format also gives superior assay reproducibility in comparison to cells in culture, since all cells are cryopreserved at the same passage number.

Accelerating your drug development program

Having the right tools for assessing your drug is key to accelerating your path to market. 

Both FDA and EMA recommend the use of bioassays during drug development:

Generally FDA considers that bioassays are more reflective (than competitive ligand-binding assays) of the in vivo situation and are recommended” (Guidance for Industry: Assay Development for Immunogenicity Testing of Therapeutic Proteins, issued by FDA, Dec 2009.)

Ideally, the potency assay will represent the product's mechanism of action (i.e., relevant therapeutic activity or intended biological effect)” (Guidance for Industry: Potency for cellular and gene therapy products, issued by FDA, Jan 2011)

For most biological products, the most appropriate neutralizing antibody assay is a bioassay which measures the neutralization of the bioactivity” (EMA Guidance on immunogenicity assessment of monoclonal antibodies intended for in vivo clinical use, issued by EMA, 2012.)

Assay Precision

The Assay Ready Format not only reduces the assay turnaround times and amount of labor required, but also reduces assay variability in comparison with cells in culture. Traditionally, cell-based assays are known for their high variability, with %CV often over 25%. Robustness assays of iLite assays have shown that repeatability and intermediate precision are in the range of 4-11% CV, and sample accuracy between 92-107%. In other words, by using iLite Assay Ready Cells, you are getting the biological relevance of a cell-based assay with the ease of use and precision normally associated with a ligand binding assay.

Sensitivity and Specificity

The sensitivity of iLite cells is enhanced through up-regulation of key components, such as the receptor and certain signaling pathway proteins. The up-regulation of receptors also confers a higher specificity, and this is enhanced through the use of chimeric transcription factors and synthetic reporter gene promoters. These elements are used as a lock and key to transcription of the reporter gene – only the chimeric transcription factor can bind to the synthetic promoter, and endogenous transcription factors are thereby unable to trigger expression of the reporter gene. In this way, pathway cross-talk can be effectively minimized.

In need of biological
drug monitoring services?

Our Diagnostic Services offer drug monitoring services using iLite reporter gene cell-based assays performed by our in-house laboratory as a lab developed tests. 

In need of cell-based 
bioanalytical services?

Our Bioanalytical Services offer a range of cell-based assay services using iLite and can help accelerate and optimize your drug development program.