Podcast
Questions and Answers
What is a key strategy to minimize immunogenicity in monoclonal antibodies?
What is a key strategy to minimize immunogenicity in monoclonal antibodies?
What characteristic defines a chimeric monoclonal antibody?
What characteristic defines a chimeric monoclonal antibody?
In the context of monoclonal antibodies, what does CDR stand for?
In the context of monoclonal antibodies, what does CDR stand for?
Which of the following types of antibodies are developed due to the high immunogenic response of murine antibodies?
Which of the following types of antibodies are developed due to the high immunogenic response of murine antibodies?
Signup and view all the answers
Which clinical aspect should be included in the immunogenicity reporting of therapeutic proteins?
Which clinical aspect should be included in the immunogenicity reporting of therapeutic proteins?
Signup and view all the answers
Which cells are classified as innate immune cells?
Which cells are classified as innate immune cells?
Signup and view all the answers
What do pattern recognition receptors (PRRs) primarily recognize?
What do pattern recognition receptors (PRRs) primarily recognize?
Signup and view all the answers
What triggers the production of proinflammatory cytokines in innate immune cells?
What triggers the production of proinflammatory cytokines in innate immune cells?
Signup and view all the answers
Which of the following is NOT a pathogen-associated molecular pattern (PAMP)?
Which of the following is NOT a pathogen-associated molecular pattern (PAMP)?
Signup and view all the answers
Which of the following immune responses is primarily associated with innate immunity?
Which of the following immune responses is primarily associated with innate immunity?
Signup and view all the answers
What does a significant increase in cytokine levels over the baseline indicate?
What does a significant increase in cytokine levels over the baseline indicate?
Signup and view all the answers
Which type of immune cells are primarily responsible for assessing the immunogenicity of therapeutic proteins?
Which type of immune cells are primarily responsible for assessing the immunogenicity of therapeutic proteins?
Signup and view all the answers
What method can be used to measure cytokine production inside the cell?
What method can be used to measure cytokine production inside the cell?
Signup and view all the answers
Which cytokines are mentioned as products of dendritic cells and macrophages?
Which cytokines are mentioned as products of dendritic cells and macrophages?
Signup and view all the answers
What type of assays focus on the innate immune system's response to therapeutic proteins?
What type of assays focus on the innate immune system's response to therapeutic proteins?
Signup and view all the answers
What is the most cost-effective stage for performing immunogenicity risk assessment?
What is the most cost-effective stage for performing immunogenicity risk assessment?
Signup and view all the answers
Which method is NOT mentioned as a strategy for assessing immunogenicity risk?
Which method is NOT mentioned as a strategy for assessing immunogenicity risk?
Signup and view all the answers
What percentage of approved biologics reported clinical immunogenicity, highlighting the risk of failure in prediction methods?
What percentage of approved biologics reported clinical immunogenicity, highlighting the risk of failure in prediction methods?
Signup and view all the answers
Which of the following approaches can be used to design candidates with low predicted immunogenicity?
Which of the following approaches can be used to design candidates with low predicted immunogenicity?
Signup and view all the answers
What is a potential consequence of having a protein with high immunogenicity observed in early human studies?
What is a potential consequence of having a protein with high immunogenicity observed in early human studies?
Signup and view all the answers
What main factors can influence the immunogenic potential of a therapeutic molecule apart from its structure?
What main factors can influence the immunogenic potential of a therapeutic molecule apart from its structure?
Signup and view all the answers
What does in vitro prediction of immunogenicity risk primarily involve?
What does in vitro prediction of immunogenicity risk primarily involve?
Signup and view all the answers
What is the purpose of performing immunogenicity risk assessment retrospectively?
What is the purpose of performing immunogenicity risk assessment retrospectively?
Signup and view all the answers
What is the primary purpose of labeling PBMCs with a proliferation dye in a CD4+ T cell proliferation assay?
What is the primary purpose of labeling PBMCs with a proliferation dye in a CD4+ T cell proliferation assay?
Signup and view all the answers
How long should PBMCs be incubated with the test article during the CD4+ T cell proliferation assay?
How long should PBMCs be incubated with the test article during the CD4+ T cell proliferation assay?
Signup and view all the answers
Why might depleting CD8+ T cells from PBMCs be beneficial in a CD4+ T cell proliferation assay?
Why might depleting CD8+ T cells from PBMCs be beneficial in a CD4+ T cell proliferation assay?
Signup and view all the answers
What does an increased incidence of T cell proliferation indicate in relation to immunogenicity?
What does an increased incidence of T cell proliferation indicate in relation to immunogenicity?
Signup and view all the answers
What role does the uptake of antigen by dendritic cells play in assessing immunogenic risk?
What role does the uptake of antigen by dendritic cells play in assessing immunogenic risk?
Signup and view all the answers
Which two antibodies are mentioned as targeting IL-17A?
Which two antibodies are mentioned as targeting IL-17A?
Signup and view all the answers
What is the significance of MHC II associated peptide proteomics (MAPPs) in immunogenicity assessment?
What is the significance of MHC II associated peptide proteomics (MAPPs) in immunogenicity assessment?
Signup and view all the answers
What does a high Incidence of anti-drug antibodies (ADA) suggest in the context of Secukinumab?
What does a high Incidence of anti-drug antibodies (ADA) suggest in the context of Secukinumab?
Signup and view all the answers
Study Notes
Humanization of Monoclonal Antibodies
- Immunogenicity is the ability of a molecule to trigger an immune response
- Strategies to minimize immunogenicity:
- Replace animal-derived drugs with human sources
- Increase human sequence content
- Change from murine (mouse) to humanized or human antibodies
- Murine antibodies are highly immunogenic
- Chimeric antibodies combine mouse variable region with human constant (e.g., Remicade, Simulect)
- Chimeric antibodies can also be immunogenic, leading to humanized antibody, where only the CDR region has murine sequences (e.g., Avastin)
- Humanization involves replacing mouse amino acids with human counterparts to minimize immunogenicity
Immunogenicity Testing in Clinical Trials
- Samples needed:
- Serum: For antibody detection
- Plasma: For cytokine and chemokine measurement
- PBMCs: For cellular assays
- Analyze samples at different time points:
- Baseline (before treatment)
- During treatment (various time points)
- After treatment
- Assays used:
- ELISA: To quantify levels of antibodies to the therapeutic protein
- Flow cytometry: To measure immune cell populations and their activation
- Cytokine assay: To assess the production of inflammatory cytokines
- T cell proliferation assay: To determine the activation and proliferation of T cells
- Report immunogenicity:
- Incidence: The percentage of individuals developing an immune response
- Titers: The concentration of antibodies in serum
- Correlation between immunogenicity and outcomes: Relationship between immune response and treatment efficacy, adverse events, and other clinical endpoints
Innate Immune Activation
- Innate immune cells recognize "self" and "non-self" through pattern-recognition receptors (PRRs)
- Examples of innate immune cells:
- Monocytes, neutrophils, macrophages, dendritic cells, natural killer cells, mast cells, eosinophils, basophils
- PRRs trigger a cascade of signals that lead to the production of proinflammatory cytokines
- PAMPs are pathogen-associated molecular patterns
- DAMPs are damage-associated molecular patterns
- PAMPs and DAMPs are conserved structures that are recognized by PRRs
Immune Cells Involved in Immunogenicity
- B cells: Produce antibodies that target the therapeutic protein
- T cells:
- CD4+ T helper cells: Amplify the immune response by stimulating B cells and other immune cells
- CD8+ cytotoxic T cells: Can directly eliminate cells that express the therapeutic protein
- Dendritic cells: Present antigens to T cells, initiating adaptive immune responses
- Macrophages: Engulf and destroy pathogens
- Neutrophils: Combat infections by releasing enzymes and other antimicrobial substances
Immunogenicity Risk Assessment
- FDA guidelines recommend immunogenicity risk assessment during drug development
- Immunogenicity is a significant factor in determining the safety and efficacy of therapeutic proteins
- Immunogenicity risk assessment strategies:
- In silico: Computational analysis of protein sequence to predict the likelihood of immune recognition
- In vitro prediction: Cell culture assays using immune cells to assess the interaction of the therapeutic protein with the immune system
- In vivo: Animal studies to evaluate the immunogenic potential of the protein in a living organism
- Clinical immunogenicity assessments: Monitoring patients in clinical trials to evaluate the occurrence and severity of immune response
- More than 89% of approved biologics have reported clinical immunogenicity, highlighting the importance of risk assessment
In Silico Screening
- Computational approaches to analyze protein sequences and identify features that might trigger an immune response
- Methods assess:
- MHC protein binding: Predication of whether protein fragments can bind to major histocompatibility complex (MHC) molecules, a crucial step in antigen presentation to T cells
- T cell epitope prediction: Identify peptide sequences within the protein that are likely to be recognized by T cell receptors
- Predictive algorithms prioritize molecules based on immune response likelihood
- These tools are valuable in early stage development to select candidates with reduced immunogenicity risk
In Vitro Immunogenicity Testing
- Cell culture methods to study the interaction of the therapeutic protein with immune cells
- Commonly used cells types:
- PBMCs: Peripherial blood mononuclear cells containing a variety of immune cells
- Dendritic cells: Present antigens to T cells
- Macrophages: Phagocytic cells that can engulf and process antigens
- Assays:
- Cytokine release: Incubate test article with immune cells and measure cytokine production in the media or inside the cell
- T cell proliferation: Assess the ability of the therapeutic protein to stimulate T cell growth and proliferation
- These assays provide insights into the immune response at the cellular level and help identify potential immunogenic risks
Factors Influencing Immunogenicity
- Molecule structure:
- Amino acid sequence: Differences from human proteins can trigger immune recognition
- Post-translational modifications: Changes to the protein after synthesis, like glycosylation
- Dosage and route of administration
- Patient factors:
- Genetics: MHC genes variability
- Previous exposure to similar proteins
- Underlying immune status
Cytokine Assays
- Measure production of cytokines, small signaling molecules that mediate communication between immune system cells
- Common cytokines assessed:
- TNF-α: Tumor necrosis factor alpha, a proinflammatory cytokine
- IL-6: Interleukin 6, a proinflammatory cytokine
- Cells often tested include:
- Dendritic cells
- Macrophages
- A significant increase in cytokine levels compared to baseline may indicate immunogenic risk
T Cell Assays
- Investigate the activation of T cells using proliferation assay or by measuring cytokine production and cell surface marker expression
- Techniques:
- Proliferation assay: Quantify the ability of the therapeutic protein to stimulate T cell division
- Flow cytometry: Analyze T cells surface markers to determine activation status
- Depletion of CD8+ T cells from PBMCs is common to enhance the assay sensitivity
MHC II Associated Peptide Proteomics (MAPPs)
- Identify the peptides that are bound to MHC II molecules, which are expressed on antigen-presenting cells like dendritic cells
- MHC II binding: Determine the peptides from the therapeutic protein that are likely to be presented to T cells, a key step initiating an adaptive immune response.
- MAPPs assess the potential of a protein to interact with MHC II and trigger an immune response
- Predictive data can guide the development of proteins with less immunogenic potential
Immunogenicity Assessment in a Novel Monoclonal Antibody Development
- In vitro testing:
- Stimulate immune cells (e.g., PBMCs, dendritic cells) with candidate monoclonal antibodies
- Measure cytokine production (e.g., TNF-α, IL-6) and T cell proliferation
- Report immunogenic potential:
- Compare cytokine levels and T cell proliferation between control and candidate molecule stimulated cells
- Evaluate statistically significant differences
- Calculate immunogenic potential based on established metrics
- Factors besides structure:
- Dosage
- Route of administration
- Patient’s immune status
Studying That Suits You
Use AI to generate personalized quizzes and flashcards to suit your learning preferences.
Related Documents
Description
Test your knowledge on the humanization of monoclonal antibodies and their immunogenicity. This quiz covers strategies to minimize immune responses, the role of chimeric antibodies, and testing methods used in clinical trials. Perfect for students in pharmacology or immunology courses.
