IMMUNOTHERAPY ASSAYS

IMMUNOTHERAPY ASSAYS Custom solutions for immunotherapy drug development.

From idea to IND/NDA/BLA – Customized services and license free assay development for immunotherapy drug development

Immunotherapy is a biological therapy method of treating diseases by activating or suppressing a person’s own immune system. Various types of human diseases such as cancer, autoimmune diseases, allergies, graft rejection (transplant rejection), neurological diseases and viral diseases are treated with immunotherapy.

Types of immunotherapy methods

1. Immune checkpoint inhibitors (Fig. 1)

2. Adoptive Cell Therapies.

• Tumor-infiltrating lymphocyte (TIL) therapy
• Chimeric antigen receptor T (CART)-cell therapy
• Natural killer (NK) cell therapy

3. Monoclonal antibodies

• Recombinant monoclonal antibodies
• Antibody drug conjugates
• Single chain antibodies, including bispecific antibodies

4. Oncolytic virus therapy

5. Cancer vaccines

6. Immune system modulators

• Cytokines (e.g., interferon-alpha (IFN-α), interleukin-2 (IL-2), etc.,)
• Bacillus Calmette–Guérin (BCG)
• Immunomodulatory drugs

Fig. 1: Currently used immunotherapy methods

Marin Biologic Laboratories (“Marin Bio”) offers customized R&D, pre-clinical and clinical assay services for immunotherapy development. Our customized services also include development of project-based innovative solutions or technologies for our clients. Our clients own the technologies we developed for them.

Immunotherapy discovery, pre-clinical (non-GMP) and GMP assays we offer:

Molecular Biology & Protein Expression

• Antibody cloning
• Recombinant antibody expression, and purification
• Biochemical and functional characterization of antibodies

ELISA/ Immunoassays

• ELISA
• Neutralizing antibody bioassays
• Pharmacokinetics (PK) assays
• Pharmacodynamics (PD) assays

Cell-based assays

• ADCC, Antibody-Dependent Cellular Cytotoxicity bioassays
• Drug or target-specific functional assays
• Reporter assays
• Flow cytometry
• CDC or CMC, Complement-Dependent Cytotoxicity or Complement Mediated Cytotoxicity bioassays
• CTL, Cytotoxic T-Lymphocyte killing bioassays
• Flow Cytometry bioassays for proliferation and surface marker differentiation
• MLR, Mixed Lymphocyte Reaction bioassays
• ADA, Anti-Drug Antibody inhibiting drug action (also see ELISA page)
• Monocyte/macrophage and granulocyte activation bioassays
• Oxidative Stress and Nitrosative Stress bioassays

Cancer immunotherapy T-cell assays

• Cytokine response assays
• T-cell proliferation assay
• T-cell killing assay
• Mixed Lymphocyte Reaction (MLR) assay
• Treg suppression assay

Immunogenicity assay

• Anti-drug antibody (ADA) assay
• Neutralizing antibody (NAb) assay
• T-cell assays: Peripheral blood mononuclear cell (PBMC)- and dendritic cell (DC)- based assays.
• Cytokine Release Syndrome Assay
• Natural Killer (NK) Cell Assay
• Phagocytosis Assay

Custom stable cell line development

• For assay development, stability testing , and lot release testing.

Master cell bank

• Production and characterization master cell bank for immunoassay, cell-based assays, and vector production.

Lot release assays/tests for immunotherapy drugs

This service includes regulatory compliant (GLP/cGMP) assays for potency lot release testing, bioanalysis, characterization, and stability.

Table 1: Immunotherapy drug release testing services

Stability assays/tests for immunotherapy drugs

• Cell Based Assays
• ELISA & other Antibody Based Assays
• Flow Cytometry and Cell Characterization
• Protein, Enzyme and other Biochemical Assays
• Immunoassays
• Radioimmunoassay
• Mixed Lymphocyte Reaction Assays (MLR)
• PCR and qPCR

Case study: Case study Functional characterization of recombinant bispecific antibody with an in-house developed tumor cell killing assay

Goal: Cell-based potency assay using an Immunogene therapy drug which binds a killer T-cells to the cancer cell line and measures cancer cell killing. This experiment compares Reference Standard to Test Sample (bifunctional or bispecific antibody construct).

The effector and target cell lines are incubated at a specific effector cell-to-target cell concentration. The target cancer cells are pre-loaded with a fluorescent dye. Increasing concentrations of bispecific antibody are added. Incubation of cells with drugs is 2.5 hours and the fluorescence released into the supernatant from dead target cells is measured (Fig. 2).

Fig. 2: Cell-based potency assay using an Immunogene therapy drug

Recent Blogs

• Single-dose immunogene therapy – A universal alternative for CAR-T cell therapy
• Capsid engineering of adeno-associated virus (AAV) vectors