All biological therapeutics have the potential to induce an immune-mediated response ranging from benign to severe adverse effects. It is important to assess the immunogenicity risk of potential biotherapeutics in producing neutralizing and non-neutralizing anti-drug antibodies (ADA), especially in clinical phases of drug development. The effects of these neutralizing and non-neutralizing anti-drug antibodies can encompass diminished clinical efficacy of the biotherapeutic being administered to hypersensitivity, allergic reaction or even cytokine storms. Anti-drug antibodies induced by biologic therapeutics often impact drug pharmacokinetics, pharmacodynamics response, clinical efficacy, and patient safety.
Immunogenicity testing on the Single Molecule Counting (SMC™) technology platform
SMC™ technology can support all phases of immunogenicity testing using digital counting on the SMCxPRO™ high-sensitivity instrument for low-level protein detection.
SMC™ advantages include:
Ultrasensitive down to pg/mL detection for low-affinity ADA and reduced need for dilutions
Wide dynamic range for detection of high-affinity ADA with minimal matrix interference
All ADA subtypes can be detected including IgM and IgE
Tolerance to high drug concentrations in sample
Reduced wash steps for detection of low-affinity antibodies and reduced assay time
Integrated software package that is 21 CFR part II compliant and LIMS compatible
SMC™ Immunogenicity Products and Services
Our assay development kit is designed to guide and assist you with the process. Our experienced application scientists can also help design, validate and also train end-users to fully utilize the platform.
SMC™ Immunogenicity Assay Development Kit (Cat. No. 03-0175-00)
Combining our immunoassay portfolio to study the impact on the immunogenicity of a therapeutic can provide great insight into the mechanism of the response. The SMC™ technology offers increased sensitivity which may assist in the detection of low-affinity antibodies and lead to earlier detection of a primary ADA response, overcome matrix effects, and may reduce drug tolerance.
Using the Luminex® platform, our MILLIPLEX® configurable complement and immunoglobulin panels can help you further elucidate the mechanism of response and the immune complex meditated responses to the ADA.
The benefits of ultrasensitive immunogenicity testing
Some products are given in low/sub nM concentrations and ADA levels of 10 ng/mL can be expected to clear or neutralise all of the given drug. The extra sensitivity of the SMC™ platform can detect ADA that would otherwise be missed by traditional technologies. By having the option of increasing dilution and still detecting ADA, drug tolerance can be improved by overcoming traditional sensitivity limitations. The drug tolerance at 100 ng/mL can sometimes be poor. The SMC™ Technology allows you to detect ADA which would previously been undetectable. Low sample volumes in mouse studies or difficult to obtain human matrixes, such as spinal fluid, can be diluted for additional assays. The enhanced sensitivity of the SMC™ technology detects both IgE, present in very low concentrations, and IgM isotypes, traditionally difficult to detect.
FDA immunogenicity guidelines
The FDA has provided guidance for the industry which sets out recommendations to facilitate the industry’s development and validation of assays for assessment of immunogenicity. In the FDA 2019 Guidance, it is recommended that the initial screening assay be able to detect all relevant immunoglobulin (Ig) isotypes. For non-mucosal routes of administration and in the absence of a risk of anaphylaxis, the relevant ADA isotypes are IgM and IgG. For mucosal routes of administration, IgA isotype ADAs are also relevant. Also, for therapeutic protein products where there is a high risk for anaphylaxis or where anaphylaxis has been observed, results from antigen-specific IgE assays may be informative.
It is recommended that screening and confirmatory IgG and IgM ADA assays achieve a sensitivity of at least 100 nanograms per millilitre (ng/mL), although a limit of sensitivity greater than 100 ng/mL may be acceptable depending on risk and prior knowledge. Recent data suggest that concentrations below 100 ng/mL may be associated with clinical events (Plotkin 2010; Zhou et al. 2013). Assays developed to assess IgE ADA should have sensitivity in the high picograms per millilitre (pg/mL) to low ng/mL range.