Linking mechanism of action (MoA) information from cell-based functional assays of increasing physiological relevance with binding data, allows the triaging of drug candidates with the most desired characteristics. Our in vitro primary cell and co-culture assay platforms are carefully designed to balance translational relevance with sample throughput, providing the level of data you need to characterise, and progress your leads with confidence. Using our deep understanding of the biology of your therapeutic target, we will select the best in vitro model to generate a complete mechanistic profile and can couple this with supporting immuno-toxicology data. 

In vitro pharmacology and extended MoA characterisation

Immune cell activation assays; T cell, NK cell, DCs, MLRs, antigen recall

Our primary immune cell assays are designed to facilitate extended MoA characterisation, support the development of your in vitro pharmacology data package and inform for potential biomarkers. For these purposes, we have developed a comprehensive suite of assay platforms employing human immune cells. 

A key feature of our comprehensive range of immunology assays and associated tools, is the high content information that can be generated in a single assay run. All primary cell assays are adapted to 96 well format and we couple this with extensive readouts to provide unparalleled phenotyping and characterisation using:

• Multi-parameter flow cytometry (up to 25 colours) using the Novocyte
• Multiplex cytokine analysis (up to 40 analytes) using Magpix Luminex or LegendPlex
• Kinetic assessments of cell viability or cytotoxicity using the xCELLigence RTCA 

We also provide access to primary patient material via our relationship with local biorepositories. Provision of ex vivo patient-derived models combined with high content readouts can validate mechanistic information generated from in vitro models and provide insight into potential biomarkers that can be used to guide inclusion criteria and design of clinical studies.

• T cell assays – T cell activation assays, including pre-screened MLRs, pre-screened antigen recall assay, polyclonal T cell stimulation; In vitro T cell exhaustion model; Regulatory T cell assays - Inducible Tregs (iTregs), natural Tregs (nTregs) and expanded Tregs (eTregs); T cell mediated target cell cytolysis (TDCC) assays employing multi-parameter endpoint or kinetic readouts
• Macrophage and dendritic cell assays – Phagocytosis (ADCP); differentiation & polarisation (M0, M1, M2a, M2b, M2c, M2d, TAMs); macrophage mediated T cell activation, MLRs 
• Natural Killer (NK) cell assays – Antibody-dependent cellular cytotoxicity (ADCC), NK cell-mediated cytolysis - immune cell engagers and NK cell therapies; NK cell activation 
• Assays employing patient-derived material – Our access to patient-derived samples, including blood-based products, allows us to assess your molecules in a more physiological relevant setting, linking disease phenotype to your molecule’s MoA. Our high content analyses (multi-parameter flow cytometry and multiplex cytokine) facilitate in depth interrogation of MoA and allow for prediction of potential biomarkers of efficacy.

Explore our interactive tumour microenvironment to learn more about the primary immune cell platforms we have available.

Antibody effector function assays

For antibody-based drugs, lead optimisation activities such as humanisation, affinity maturation and Fcγ receptor engineering can have a significant impact on binding and effector functions. Our bioanalysis experts provide accurate and robust solutions that allows fine resolution of the effects of these activities on binding, efficacy and effector functions:

• Binding assessments – Antigen, Fcγ receptor binding, C1q binding and dependent binding for multi-specific molecules using various methodologies including SPR, homogenous assays (AlphaLisa and AlphaScreen), ELISA and cell-based binding by flow cytometry
• Signaling reporters – large portfolio of in-house developed signaling reporter systems, including Fcγ reporter reporters, in 384-well plate and “Thaw&Go” formats
• Effector function characterisation – ADCC, ADCP, trogocytosis, CDC, T-cell mediated target cell cytolysis, employing pre-screened or genotyped donors, in 96- and 384-based formats to reduce variability and increase assay accuracy and robustness.

Employing the correct effector cell preparation is critical and the choice should be influenced by the assay methodology and the hypothesis under test. We take several steps to enhance assay performance by: 

• Providing direct access to characterised PBMC and whole blood donors (CD16a genotyped; V/V, V/F, F/F, CD32a genotyped R/R, R/H, H/H)
• Stocking thousands of cryopreserved PBMC vials
• Performing functional pre-screening assays for ADCC (As well as TDCC, MLRs and antigen recall) 
• Preparing isolated immune cell banks.

In vitro immuno-toxicology

Cytokine Release Syndrome (CRS), Tissue cross-reactivity (TCR) and “on-target, off-tumour” hazard assessment

In vitro hazard assessments are a regulatory requirement for many therapeutics, particularly targeted biologics. We have several discrete assay formats that we can apply to your molecule to reduce the risk of unanticipated results at the expensive clinical trial stages.

• Measurement of “On-target, Off-tumour” effects – Antibody Analytics has developed proprietary cell line platforms that facilitate assessment of the impact of target antigen density on potency and efficacy of a targeted therapeutic. This allows the estimation of the impact of antigen density (e.g. number of receptors) on a molecule’s therapeutic index. Contact our team to learn about our system 
• Cytokine Release Assay (CRA) – We test therapeutics in plate bound and soluble formats, comparing whole blood with PBMCs directly and benchmark against approved antibodies. We consider the MoA of your therapeutic when designing the assay. For therapies such as immune cell engagers and engineered cell therapies, cytokine release can be part of the intended mechanism of action, and this is considered in both the design and interpretation
• Identifying antigen cross-reactivity – Tissue cross-reactivity (TCR) studies are a regulatory requirement for targeted therapeutics and in collaboration with our partners we can provide access to GLP-accredited TCR assessment in primary human tissues, performed and presented in a format acceptable to regulatory bodies and in compliance with their requirements.

Custom assay development 

Our scientists relish the opportunity to establish novel cell models and platforms. With all of our experts under one roof, they can readily brainstorm and collaborate to provide the solutions you need. Speak to one of our biology experts today to discuss the unique challenges you are facing.