Immunotoxicology Assays For Early Immune Safety Assessment

In vitro cytokine release assays are essential immunotoxicology assays used to evaluate the potential of drug candidates to trigger excessive immune activation, including cytokine storms. These events represent a serious safety risk and are a recognised regulatory concern, particularly for biologics and immune-modulating therapies.

RoukenBio offers advanced cytokine release assays, including PBMC cytokine release assays, designed to generate clinically relevant insights into immune activation risk. Our assays support early identification of safety liabilities, helping teams refine candidates and reduce downstream development risk.

What sets our cytokine release assays apart:

• Well-characterised donors
   Highly characterised donor panels improve reproducibility and relevance when assessing immune activation risk.
   
• High-content cytokine analysis
   Multi-parameter cytokine profiling provides a detailed view of inflammatory responses beyond single-analyte readouts.
   
• Tailored assay designs
   Assays are customised to your therapeutic modality, target biology, and mechanism of action, with soluble and plate-bound formats available.
   
• Multiple assay formats
  Extensive experience across:
  
  PBMC cytokine release assays
  High-density PBMC formats
  Whole blood assays
   

By integrating high-content analysis with flexible PBMC-based assay formats, RoukenBio delivers actionable immunotoxicology data that supports safer candidate progression and more confident clinical decision-making.
 

On-target, off-tumour side effects represent a significant immunotoxicology risk during the development of antibody-targeted therapies for solid tumours. This includes immune cell engagers, CAR-T therapies, antibody-drug conjugates (ADCs), and conventional monoclonal antibodies. While regulatory authorities such as the FDA increasingly emphasise early evaluation of these risks, predictive in vitro models have historically been limited.
 

RoukenBio addresses this challenge through its proprietary IndEx-2 system, a specialised in vitro platform designed to predict and characterise on-target, off-tumour toxicities with a high degree of control and translational relevance.
 

Key features of the IndEx-2 system include:
 

• Broad therapeutic applicability
  Suitable for immune cell engagers, cell therapies, ADCs, and monoclonal antibodies across multiple tumour indications.
   
• Precision antigen control
  A finely titratable, inducible antigen expression system enables independent control of one or two receptor expression levels within a consistent genetic background.
   
• Multi-mechanism impact analysis
  High-content methodologies allow assessment of how antigen density influences multiple mechanisms of action, supporting a deeper understanding of efficacy and safety trade-offs.
   
• Correlated tissue relevance
  In vitro findings are contextualised using primary tissue expression data to strengthen clinical interpretation.
   
• Flexible cell line options
  Access to a broad panel of off-the-shelf cell lines, with the option to develop custom models tailored to your target biology.

By integrating the IndEx-2 system into immunotoxicology assessment strategies, RoukenBio enables earlier identification of off-tumour risk, supporting safer candidate selection and more informed progression decisions.
 

Immunogenicity is a critical component of immunotoxicology assessment, particularly for biologics and advanced therapies where unintended immune responses can compromise safety, efficacy, or long-term dosing. RoukenBio provides a comprehensive suite of immunogenicity assays designed to assess how therapeutic candidates interact with the immune system and to identify risk factors early in development.
 

Our immunogenicity assessment services support mechanistic understanding, candidate optimisation, and regulatory-aligned evaluation across multiple immune pathways.
 

Our immunogenicity services include:

DC Maturation Assays:

• Utilise monocyte-derived dendritic cells (moDCs) to assess the potential of your therapeutic to induce ADA (anti-drug antibodies).
• Measure cell surface marker expression (e.g., CD83, CD80, CD86) via flow cytometry.
• Analyse cytokine secretion (e.g., IL-1β, IL-6, TNFα) and signalling pathway activation (e.g., AkT, Syk).
• Tailored to assess formulation factors or protein aggregation that may contribute to immunogenicity. 
   

T Cell Activation Assays:

• Evaluate immunogenic risk by measuring the activation of CD4+ T cells in response to therapeutic proteins.
• Multiple assay formats available, including PBMC-based assays, DC-T cell co-culture systems, and CD4+ T cell-specific setups.
• Assess key readouts such as early T cell activation markers (e.g., CD154, CD137), cytokine secretion (e.g., IL-2, IFNγ), and T cell proliferation (e.g., Ki-67 expression). 
   

ADA Assay Development:

• Method development and validation for assessing anti-drug antibodies (ADA), aligned with EMA and FDA guidelines.
• Tiered strategies for binding and neutralising ADA, including determination of antibody class and titre. 
   

Key Benefits of Our Immunogenicity Assessments:

• HLA-characterised donor cohort: Our donor pool has an HLA distribution corresponding to the world population, and can be tailored to specific subpopulations of interest.
   
• Multiple readouts: We offer a variety of readouts to assess different modes of immunogenic potential, ensuring comprehensive evaluation.