Immunobiology Bioanalysis

Agilex offers a comprehensive immunobiology service to enable sponsors in the drug development arena to understand how their therapeutic is interacting with certain cell types; which cell types are being modulated by the therapeutic and which pathways and effector genes or proteins are being modulated. With extensive experience in autoimmune diseases, immunology and inflammation, Agilex is the CRO partner of choice for high quality results. Agilex’s Immunobiology services include multicolour flow cytometry services (up to 20 colors), droplet digital and conventional PCR assays, Elispot/Fluorospot, PBMC isolations and custom whole blood cell based assays.

Flow Cytometry Services

Flow cytometry is a powerful tool used in clinical studies to investigate the impact of new drug substances on immune cell function, providing critical information for safety and efficacy. By using antibodies to cell surface or intracellular proteins tagged with fluorescent markers, flow cytometry enables the rapid collection of data related to multiple parameters for a heterogenous cell mixture, in a single tube. Flow cytometry is thus key to understanding how a drug substance has altered the profile of cells or their function.  Agilex Biolabs has state-of-the -art flow cytometry facilities, boasting custom built BD FACSymphony A3 flow cytometers, featuring 5 lasers and 20 colours to facilitate investigations into immune cell function.

Flow Cytometry assays to support any phase of drug development.

• Immune Cell Profiling

• Cellular function Assays (eg: cellular proliferation and intracellular signaling) 

• Receptor Occupancy Assays

Immune Cell profiling

Immune cell profiling involves the analysis of immune cells, including T-cells, B-cells, NK-cells, monocytes, macrophages and dendritic cells. Broadly referred to as immunophenotyping, immune cell profiling provides powerful multiparametric analysis to distinguish various populations of cells and the impact of a drug substance on them. At Agilex Biolabs, we have a ready to use off the shelf TBNK multitest assay (BD Biosciences), which is IVD/CE designated and clinically useful for monitoring T, B and NK cells during a clinical study. The assay can measure relative abundance, but also absolute numbers when combined with counting beads. Immune cell profiling is usually best performed using fresh samples, due to the instability of certain cell populations. However, this can often be overcome for specific cellular markers using specialized collection tubes which contain a stabilizer or fixative (eg: Transfix tubes). Agilex recently conducted a comparative study between K2 EDTA and Transfix tubes to determine the precision of both post-stain and sample stability of fresh and fixed CD3, CD4, CD8, CD19 and NK cells in whole blood with and without fixation. Our study showed that Transfix tubes could extend sample stability to 10 days.

See the results of our study here (link to TBNK technote).

Cellular function Assays

T cell and B cell function is critical to adaptive and humoral immunological responses to infection and disease (eg: cancer and auto-immune diseases). Flow cytometry is a powerful tool to enable clinical investigators to identify the expression and functionality of specific populations of immune cells in a rapid and multiparametric manner. At Agilex Biolabs our Scientists have extensive experience in developing custom flow cytometry assays to investigate the impact of drug substances on immune cell function. Our Scientists recently conducted a study to investigate the precision of T cell proliferation and marker analysis in cultured polymorphonuclear (PBMC) cells by flow cytometry. The study, which can be found here (link to T cell technote), provides powerful information on the influence of cell culture on assay precision, particularly when using cell permeable dyes for proliferation analysis.

Over the past decade, clinical studies featuring Immuno-modulator therapies, either antibody drugs or small molecule chemical inhibitors of key signaling pathways, have increased considerably. Key to identifying efficacy of these drug substances are Flow Cytometry assays measuring activation or phosphorylation of signal transduction proteins as many of these novel therapies are designed to inhibit pathways and inhibit phosphorylation. Agilex recently conducted a study to investigate the efficacy of a novel anticancer drug. Part of the efficacy study was the development of an intracellular staining phospho STAT Flow Cytometry assay and droplet digital PCR assay. Using a one-step fix, lyse, permeabilise method, Agilex Scientists successfully setup up the method to support the clinical study. Read more about the assay and how Agilex can support your drug development program, here (link to pSTAT and ddPCR technote).

Receptor Occupancy Assays

Receptor occupancy assays are instrumental in discerning the interaction between biotherapeutics and their target receptors. These assays delve into the intricacies of drug-receptor binding kinetics. They shed light on critical parameters such as the duration of drug-receptor interaction and the degree of receptor saturation. By elucidating the pharmacodynamic profile of a drug candidate, receptor occupancy studies furnish invaluable insights into its mechanism of action and therapeutic potential. At Agilex Biolabs, our Scientists have decades’ experience in developing Receptor Occupancy assays – either free or total assays – to support clinical trials. Contact us today to discuss how Agilex can support your drug development program.

Droplet digital and Conventional PCR Assays

Our expert scientific team offers a breadth of molecular biology services in the development, optimization and validation of droplet digital or PCR/qPCR assays to support preclinical and clinical stage drug development. Our laboratories are equipped with the latest generation of PCR equipment including BioRad’s droplet digital PCR suite (CFX96 thermocycler, BioRad CX200 droplet reader) as well as a Revvity Chemagic Liquid handling robot to support biodistribution and shedding assays for novel oncolytic virus programs, cell and gene therapy programs or oncology studies. Our team recently set up a circulating tumor DNA (ctDNA) project for the detection of single base pair mutations to detect an inactivating mutation in codon 600 of exon 15 of the Braf gene using the Biorad droplet digital PCR system. The study (which can be downloaded here) demonstrates the sensitivity and precision of the BioRad System to enable detection of low copy number singe point mutations, critical for investigators within the immune-oncology drug development space. Contact us today to discuss how Agilex can support your next PCR project.

PCR Assays to support preclinical and clinical drug development

• Custom ddPCR/qPCR assays

• Gene expression analysis

• Mutation detection

• Biodistribution/shedding

• PK of RNA therapeutics

ELISpot/Flurospot

Agilex offers Enzyme-linked Immunosorbent Spot (ELISpot) assays to enable the enumeration of single antigen producing immune cells with high sensitivity in either single assay or multiplex (Flurospot) format. Agilex’s immunobiology team have extensive experience in cell enrichment assays using magnetic activated cell sorting (MACS) to support a range of ELISPot programs in the immune-oncology or vaccines arena, or enrich the cellular population down to the cell type of interest to support flow cytometry. The Agilex team recently conducted a study to analyse discrete antigen producing lymphocyte populations from whole blood using MACS-isolation, ELISPot and flow cytometry (available to download here). To learn more about how Agilex can support your next program, contact our scientists for a confidential discussion today. 

PBMC isolations and custom whole blood cell-based assays

Peripheral Blood Mononuclear cells (PBMCs) are powerful tools in multicenter immuno-oncology and vaccine clinical trials and coupled with flow cytometry or ELISpot, provide vital information on immune function – either associated with a therapeutic, or as a consequence of its role in modulating immune function. The Agilex scientific team have experience in isolating PBMCs from whole blood and have a network of laboratories across the phase 1 units in Australia to facilitate fast preparation and storage of PBMCs.Agilex team recently conducted a longitudinal study to investigate PBMC stability to support clinical studies (PBMC stability Technote) in Australia In addition, the Agilex team have experience in MACS-cell separation, stimulation and activation assays to support cytokine release (technical note available here) of immune cell function (proliferation and Activation) (technical note available).