iLite® T-cell Activation Bioassays 

A Biological Relevant and powerful way to assess CD3 induced T-cell activation

iLite® CD3 Effector Cell/ T-Cell Activation Bioassays

The iLite CD3 Effector Cell line is a reporter gene cell line that offers a convenient and powerful way of measuring the efficacy antibodies have to elicit  T cell activation through CD3  interactions. 

In recent years, a variety of immunotherapy strategies aimed at inducing, strengthening, and/or engineering T cell responses have emerged as promising approaches for the treatment of diseases such as cancer and autoimmunity. With the iLite CD3 Effector cell -  a cleverly engineered effector cell that closely resembles the natural engagement of the TCR/CD3 complex, and homologous target cells with a controlled antigen expression (+) or depletion (-),  this can be assessed with a robust assay.

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The iLite  T-Cell Activation Bioassays is a dual cell system based on bioluminescent cell-based assays. The portfolio consists of cell lines provided in an “assay-ready” format for a rapid and convenient workflow and a further reduction in assay variability and can be used for the discovery and development of novel biologics such as bispecific antibodies and CAR-T cell therapies

 

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T-Cell Activation in vivo
A mechanism of cell-mediated immune defense

T cells play a central role in cell-mediated immunity and can mediate long-term, antigen-specific, effector, and memory responses. 

T cell activation requires activation of two signals in T cells: recognition of antigen through the TCR/CD3 complex and CD28 co-stimulation via CD80/CD86. Co-engagement of these receptors on the cell surface leads to intracellular signaling events and the activation of nuclear transcription factors such as Nuclear Factor of Activated T cells (NFAT), NF-kB, and AP-1.

ILITE T-CELL ACTIVATION ASSAY - How does it work?

To be activated, the T-cells must receive several signals through antigen-presenting cells and engagement of the CD3/TCR Complex. Additional signals are provided by co-stimulatory molecules and their interactions. Many bi-specific antibody-based therapeutics utilize this activation pathway to induce a targeted immune response from the body's own T-cells.  The iLite T-cell activation product line offers a convenient and powerful way of measuring the efficacy of antibodies to elicit this effect in vitro.  

Luminescence is based on the crosslinking of CD3 complex, antibody, and target

The T-cell activation process is triggered when the effector cell interacts with a drug antibody bound to its target cell. The CD3 complex on the surface of the effector cell binds to the CD3 part of the antibody, thus creating a bridge between the effector and target cells.

Following the formation of this bridge, the engineered effector cells will, instead of activating the T-cell as in the in vivo situation, produce luciferase through an intracellular pathway and generate luminescence exclusively from this cross-linking and signaling. The strength of the luminescence correlates to the ability of the drug to activate the T-cells. 

A normalization gene for easy comparisons between samples

Like most iLite cell lines, the iLite CD3 Effector Cells also have a secondary luciferase readout from a luciferase gene expressed under the control of a constitutive promoter. This enables normalization of each individual readout according to cell number and compensates for potential matrix effects.

 

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APPLICATIONS for the iLite® ADCP Portfolio

Novel biologics formats, including bispecific antibodies and CAR-T cell therapies, require robust, reproducible measurement of T cell activation.

The iLite CD3 T-Cell activating  Reporter Bioassay is designed for evaluating CD3 mediated T-cell activation, exhibits greatly reduced variability, and offers an easier workflow than traditional T-cell activation Assays. 

 Applications such as

  • Potency lot release and
  • Antibody screening
  • Assessing the comparability between innovators and biosimilars
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TARGET CELL PORTFOLIO

Engineered, homologous, and specific target cells that enable screening for antibody induced effector functions

Can´t find a perfect match? Let us develop it for you!

In case you haven't got an ideal target cell line and can't find what you’re looking for in our off the shelf solution, you can rely on our extensive expertise to develop the most appropriate one and we have vast experience in designing cell lines ideally fit for purpose!

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