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Flow Cytometric approach to evaluate biotherapeutics' PharmacoDynamic
At ABL, we have already developed and qualified methods to assess the PharmacoDynamics analyses of therapeutic monoclonal antibodies within the framework of clinical trials. Today, we share data obtained from a Receptor Occupancy assay.
The principle of this assay is simple - as shown below - although mastering the method remains fairly complex.
The precision of the method developed at ABL has been carried out through multiple operators and instruments during several days.
The method was then used to measure the Receptor Occupancy during the course of multiple phase I/II clinical studies at different days (D0, D1, D7, D14, D21 and D28) in patients treated with low (A) or high (B) dose of the therapeutic mAb.
Assess your Immune Checkpoint Modulator
ABL is an expert in ranking drug candidates that modulate Immune Checkpoint Inhibitors.
Recent success of marketed molecules targeting immune checkpoints involved in T cell responses, such as Yervoy (Ipilimumab, anti-CTLA4), Keytruda (Pembrolizumab, anti-PD1) and Opdivo (Nivolumab, anti-PD1), in cancer patients have revived the field of immunotherapeutic approaches in oncology and more particularly of Immune checkpoint modulators.
At different levels of the immune system, as soon as an activation signal is delivered, inhibitory mechanisms get engaged to contain adverse effects of an exacerbated immune response (e.g. inflammatory chronic diseases, autoimmunity….).
For immune checkpoint inhibitors targeting innate immune responses, and more specifically NK cells, ABL has developed and validated an assay to evaluate the cytotoxic function of NK cells by flow cytometry, measuring the CD107a degranulation and the capacity of NK cells to produce IFN-gamma in response to target cells (Figure 1). In this example, the candidate antibody targeting an inhibitory receptor enhances the cytotoxic function, as demonstrated by increased frequency of CD107a/b positive cells and of IFN-g secreting cells. This representative example illustrates the low variability of the assay performed by 2 different operators in 5 independent series of experiments on one reference sample.
For immune checkpoint inhibitors targeting adaptive T cell responses, his scenario can be mimicked in an in vitro Mixed Lymphocyte Like Reaction (MLR), in which CD4+ T cells, when co-cultured in the presence of allogeneic dendritic cells, will get activated. In this model, ABL can evaluate the capacity of test compounds targeting immune check points to modulate proliferation or cytokine production, as shown below.
ABL is the immunomonitoring partner of several ongoing preclinical studies and clinical trials.
You can get a first taste of our approach for method validation and immune marker testing by downloading our application notes.
We’ve chosen to spotlight through 3 distinct case studies our contribution to:
- The functional and phenotypic characterization of immune cell subsets in the context of a Phase II clinical trial evaluating an antibody candidate
- The monitoring of the host immunological response during treatment course in a Phase II trial evaluating a candidate vaccine through the dosage of several cytokines
- The characterization of the PK/PD properties of a monoclonal antibody using a receptor occupancy assay developed and validated by ABL.
ABL produces Beads Based Critical Reagents
In order to develop specific customized assays, ABL has developed an expertise in coupling biomolecules onto several kinds of beads (magnetic, latex, or fluorescent)..
We curerently develop and produce high quality microparticle-based reagents. Our product quality relies on our track-record of analytical methods & robust quality system.
Coating microparticles can be conducted using either your specific biologics or reagents from commercial suppliers. ABL can for example develop suitable reagents for xMAP (Luminex) or Simoa (Quanterix)..
We are proud to be the provider of the world largest supplier of Spherotest microparticle-based kits. These assay kits measure the specific concentration of antibodies by an easy and rapid microagglutination technique.
A major concern for pharmaceutical companies
Biotechnology-derived proteins are increasingly used as therapeutic agents that may induce unwanted humoral and cellular immune response.
The consequences of such immune reactions to a therapeutic protein range from transient appearance of anti-drug antibodies, reduced efficacy of the drug, to severe life threatening conditions (allergy, anaphylactic reactions, induction of autoimmunity).
Therefore, immunogenicity of therapeutic proteins has to be addressed at the earliest possible stage of drug development.
Immunogenicity is the capacity to induce an immune response. Every protein-based molecule has an immunogenic potential.
While immunogenicity is seeked for vaccines, it is a serious issue for therapeutic proteins (such as antibodies, peptides, hormones, etc…).
The company is currently focusing its strategy on protein biomarkers of clinical interest and their applications for in-vitro diagnostic.
Indicia Biotechnology addresses the mutation of the pharmaceutical market in providing a wide range of customized services from biomarker validation to theranostic solutions across many therapeutic areas.
Biomarker analysis contributes to diagnose diseases, to evaluate the efficacy and the safety of drug candidates, to predict or to monitor the response of patients to medications.
Custom Assay Development
As an outsourcing alternative to internal assay design & development, we develop customized assays in multiple application fields: biomarker quantification, immunophenotyping, anti-drug antibody detection, pharmacokinetic, quality control methods,...
Validations of assays are performed following the current FDA "Guidance for Industry - Bioanalytical Method Validation" of May 2001 and/or the current EMEA “Guideline on validation of bioanalytical methods – Committee for medicinal products for human use” – November 2009.
For specific Clinical In Vitro Diagnostic tests, some specific steps are added to the validation process such as long term stability or shipment conditions.