Creative Biolabs provides detection and characterization assays for anti-drug antibodies (ADAs) of biotherapeutic drug candidates. Following the current FDA and EMEA guidelines and AAPS white papers, Creative Biolabs has developed many approaches to detect the ADA responses by using a variety of leading instrumentation platforms, such as ELISA/RIA, surface plasmon resonance (SPR) and electrochemiluminescence (ECL).

Background

Arising of anti-drug antibodies (ADAs) against biotherapeutic drugs has been widely observed in clinical applications, such as recombinant Factor VIII, and interferon beta. ADAs have been associated with the reduced clinical efficacy of biotherapeutic drugs as well as a variety of potentially serious clinical adverse effects. ADAs can be classified into two categories, neutralizing ADAs (NAbs) that block the binding of biotherapeutic drugs to their targets, and non-neutralizing ADAs (non-NAbs) that mainly contribute to faster clearance and drug resistance. ADAs have also been linked with increased side effects, such as infusion reactions and even life-threatening auto-immune syndromes caused by the cross-reactivity of ADAs with endogenous proteins. Therefore, evaluation the ADA reactions against biotherapeutic drugs in both preclinical and clinical studies is of great value for efficacy optimization and risk management of biotherapeutic drug candidates, which is required for IND application at the same time.

SIAT® Anti-Drug Antibodies (ADA) Assays

With more than ten years’ experience in providing excellent services for researchers in both academia and biopharmaceutical industry, Creative Biolabs is an experienced expert on ADA assay development and validation. We strive to accelerate the development of your new drug candidate from preclinical to clinical stage and eventually to the market. Our SIAT® anti-drug antibodies assay services include but not limit to the following.

Positive control and reagent formation

Before ADA assays, suitable positive controls must be obtained. We can prepare polyclonal or monoclonal antibody against the active ingredients that are similar to the actual ADA samples. In addition, drugs labeled with biotin or fluorescent tags can be synthesized.

Assay method development & validation

We will develop proper assays based on currently used ones such as “Bridging”, direct or competitive ELISA, cytokine profile, sensitivity, specificity, and precision evaluation and stay in compliance with FDA and EMEA guidelines and AAPS white papers.

Sample analysis

Characterizations of ADAs include the titer, isotype of ADAs, and detection of neutralization status of the ADAs. A general problem with ADA assays is the formation of drug-ADA complexes in the sample. We have developed several approaches such as pre-incubation with ADA assay reagents and acid dissociation to solve this problem.

Pharmacokinetic /Toxicokinetic (PK/TK) analysis

Evaluation of possible changes in PK/TK that are induced by ADAs.

More SIAT® Immunogenicity Related Services at Creative Biolabs

• In Silico Immunogenicity Assessment
• In Vitro Class II HLA Binding Assay
• Ex Vivo Immunogenicity Assessment
  • DC-T Cell Proliferation Assay
  • Antigen Presentation Assay
• In Vivo Immunogenicity Assessment
• Anti-drug Antibodies (ADA) Assays
• De-immunization Service

Published Data

• Anti-Drug Antibodies in Pigtailed Macaques Receiving HIV Broadly Neutralizing Antibody PGT121

Fig. 1 ADA at time of infusion negatively correlates with viral suppression by PGT121. (Wen Shi Lee, 2021)

The study investigates the effects of anti-drug antibodies (ADAs) on the efficacy of the broadly neutralizing antibody PGT121 in pigtailed macaques infected with SHIV, revealing that multiple intravenous doses of PGT121 induce ADAs which subsequently reduce the plasma concentration of PGT121 and impair its viremia suppression capabilities. Notably, these ADAs target the constant regions of the human antibodies rather than their variable domains, indicating an immune response to the structural elements of the administered antibodies. This research highlights the significant challenge ADAs pose in antibody-based HIV therapy, stressing the importance of developing methods to mitigate ADA formation to sustain the therapeutic effectiveness of monoclonal antibodies in medical treatments, which is crucial for guiding future preclinical and clinical studies on the durable efficacy of such therapies.

FAQ

1. What are anti-drug antibodies (ADAs)?

   ADAs are immune responses generated by the body against administered therapeutic proteins or biotherapeutics. These antibodies can potentially neutralize the drug's effect or alter its pharmacokinetic profile, impacting the drug's efficacy and safety.

2. Why is detecting ADAs important in biotherapeutic development?

   Detecting ADAs is critical because they can influence the therapeutic efficacy and safety of biotherapeutic drugs. By understanding the presence and impact of ADAs, developers can better assess potential adverse effects and adjust treatment protocols to improve patient outcomes.

3. What are the common techniques used for ADA detection?

   Common techniques include enzyme-linked immunosorbent assays (ELISA), electrochemiluminescence (ECL) assays, and radioimmunoassays (RIA). Each method has specific advantages in sensitivity and specificity, with ELISA being widely used for its balance of ease and effectiveness.

4. How do tiered approaches work in ADA testing?

   The tiered approach to ADA testing typically involves three steps: screening for ADAs, confirming positive results from the screening test, and characterizing the confirmed ADAs. This structured approach helps to accurately identify and understand the clinical relevance of the ADAs.

5. What is the significance of neutralizing antibody (NAb) assays in ADA characterization?

   NAb assays are crucial for determining whether ADAs can inhibit the biological activity of the drug. This information is vital for assessing potential impacts on efficacy and safety, as neutralizing ADAs can directly counteract the therapeutic benefits of the drug.

6. How do drug tolerance levels affect ADA assays?

   Drug tolerance refers to the assay's ability to detect ADAs in the presence of the drug. High drug tolerance in an assay is essential to accurately measure ADA levels without interference from the drug itself, ensuring reliable results even at therapeutic concentrations.

7. What role does assay sensitivity play in ADA detection?

   Assay sensitivity is crucial for detecting low levels of ADAs, which is important for early identification of immune responses that may later result in clinical issues. Sensitive assays help in monitoring ADA development over time, allowing for timely intervention if needed.

8. How are ADA assays validated?

   ADA assay validation involves confirming that the assay is reliable, reproducible, and suitable for its intended use. Validation parameters include specificity, sensitivity, reproducibility, and robustness, ensuring that the assay consistently provides accurate and meaningful data.

Resources

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All listed services and products are For Research Use Only. Do Not use in any diagnostic or therapeutic applications.