Immunogenicity assessment is a key element in ensuring the safety and efficacy of Antibody-Directed Enzyme Prodrug Therapy (ADEPT). At Creative Biolabs we use advanced computational models and laboratory analyses to deliver a range of immunogenicity services. Our goal is to assist clients in identifying potential immune responses and monitoring cellular immune reactions triggered by antibody drugs, thereby providing valuable data to support the subsequent optimization of ADEPT.

Introduction

Immunogenicity analysis plays a crucial role in the development of ADEPT, ensuring patient safety while also supporting the optimization of drug design and improving therapeutic efficacy. To assist clients in evaluating the safety of antibody therapeutics and their pharmacokinetic properties, Creative Biolabs has introduced a range of immunogenicity analysis services. These services aim to enhance drug design and formulation, ultimately minimizing the risk of adverse immune reactions.

• Integrated Services: We offer comprehensive immunogenicity testing services that span from the early development phase through clinical trials, ensuring consistency and complementarity of data at every stage.
• High-Throughput Testing Technology: Based on advanced high-throughput screening techniques, we can enhance testing efficiency and improve the reliability of experimental results.
• Tailored Analysis Solutions: Our scientists provide customized immunogenicity analysis plans based on the specific requirements of each project, catering to the unique characteristics of different antibody drugs.

Fig.1 The Immunogenicity of Antibody Drugs.^1,3

Services

At Creative Biolabs, immunogenicity analysis not only evaluates the immunogenicity of existing drugs but also provides valuable data to optimize antibody variants during the early stages of drug design. Furthermore, our labs can use the findings from these analyses to select protein sequences or modification techniques that exhibit lower immunogenicity, thereby reducing the likelihood of immune reactions.

• Immunogenicity Risk Assessment
  - Using advanced tools to evaluate the immunogenicity risk of antibody therapeutics, analyzing potential immunogenic sites within the drug's molecular structure.
• Antibody Response Testing
  - Conduct tests for both specific and non-specific antibody responses through ELISA to evaluate the presence of antibodies in clinical samples, thereby monitoring the immune responses of patients to the drug.
• T Cell Response Analysis
  - Employ flow cytometry and ELISPOT techniques to assess the peripheral blood T cell responses in patients to antibody drugs, to determine their immunogenic potential.
• Neutralizing Antibody Activity Testing
  - Measure the activity of neutralizing antibodies induced by the antibody drug to evaluate their impact on therapeutic efficacy.
• Long-term Immune Monitoring
  - Perform regular immune monitoring to track the dynamic changes in antibody production.

Fig.2 Immunogenicity Analysis of Antibody Drugs on Animal Models.^2,3

Procedures

STEP 1: Characterization of Drug Molecule Structure

Conduct a comprehensive analysis of the molecular structure of therapeutic antibodies, focusing on aspects such as their amino acid sequences, glycosylation modifications, and characterization of secondary and tertiary structures. These characteristics can significantly influence the drug's immunogenicity, establishing a structural framework for subsequent analyses.

STEP 2: Immunogenicity Prediction

Utilize bioinformatics assays to identify potential immunogenic epitopes. Our assays evaluate the risk of immunogenicity by determining how strongly antigenic peptides bind to HLA molecules. This process is essential for the early detection of areas that could trigger negative immune reactions.

STEP 3: Animal Model Experiments

Inject therapeutic antibodies into mice or other animal models and monitor the immune response, specifically the production of anti-drug antibodies (ADA). By designing experiments with repeated dosing, varying doses, and different administration routes, a more comprehensive understanding of the drug's immunogenicity can be achieved.

STEP 4: ADA Detection and Analysis

Develop sensitive methods for detecting ADAs, including ELISA and neutralization assays, to quantitatively and qualitatively analyze ADA production. Investigate the effects of ADAs on drug efficacy and toxicity, and monitor their variations across different doses and time points.

STEP 5: Stability and Variability Assessment

Evaluate the physicochemical stability of therapeutic antibodies and their degradation in biological environments, identifying specific conditions that may lead to alterations in antigenic epitopes. Factors such as thermal stability and pH changes can affect structural integrity, potentially increasing immunogenicity.

Case Study

Recently, Creative Biolabs has conducted an effective ADEPT Immunogenicity analysis study through a variety of systematic analysis systems.

1. Assessment of Immunogenic Risk

Initially, a preliminary analysis of the candidate antibody will be used by our labs to evaluate its potential immunogenicity. This assessment service primarily focuses on the sequence characteristics, structural features, and similarity to human antibodies.

2. Selection of Animal Models

Before conducting experiments, appropriate animal models (such as mice or monkeys) are chosen based on the characteristics of the antibody, determining whether to use immunosuppressive or non-immunosuppressive models.

3. Preparation of Test Samples

Candidate antibodies are produced and purified through recombinant DNA technology. Typically, protein A affinity chromatography is employed to achieve high-purity antibodies, ensuring the reliability of subsequent experiments.

4. Immunogenicity Testing

(1) Immunization: The purified antibody is administered to the animal model via injection at a specified dose (commonly 1 mg/kg), typically either intraperitoneally or intramuscularly.

(2) Sample Collection: Blood serum samples from the animals are collected at regular intervals after immunization—specifically at 1 week, 2 weeks, 4 weeks, and 8 weeks—to capture the dynamic changes in the immune response.

(3) Antibody Titer Determination:

ELISA is utilized to identify specific antibodies targeting the candidate antibody in the serum of test animals. A concentration gradient is created, after which serum samples are incubated with the antibody. This is followed by the introduction of enzyme-conjugated secondary antibodies, which trigger a color reaction. The resulting optical density values are then measured.

(4) Immunophenotyping Analysis:

Flow cytometry is employed to analyze the activation status of T cell subsets and B cells, assessing the nature and intensity of the immune response.

Creative Biolabs offers a collection of immunogenicity services to accelerate the success of your ADEPT projects. Through in-depth research and analysis, we assist you in evaluating and optimizing the immunogenicity of antibody therapies, ensuring their safety and efficacy in clinical applications. Feel free to contact us anytime for more information about our immunogenicity services.

References

1. Pedersen, Morten E., et al. "Assessment of immunogenicity and drug activity in patient sera by flow-induced dispersion analysis." Scientific Reports 12.1 (2022): 4670.
2. Kim, Yun Jung, et al. "Preclinical immunogenicity testing using anti-drug antibody analysis of GX-G3, Fc-fused recombinant human granulocyte colony-stimulating factor, in rat and monkey models." Scientific Reports 11.1 (2021): 12004.
3. Distributed under Open Access license CC BY 4.0, without modification.

For Research Use Only | Not For Clinical Use