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Immunogenicity overview of therapeutic biologics
- 1. Immunogenicity Overview of Therapeutic Biologics
- 2. What is Immunogenicity? The propensity of the therapeutic biologics to generate immune responses Wanted immunogenicity Unwanted immunogenicity
- 3. Background of Immunogenicity-APC Antigen + APC Phagosome Endosome/lysosome Endoplasmic reticulum MHC II Golgi apparatus <34 AA peptides T cell epitopes Secrete vesicles Peptide-MHC-II complex CD4+T Surface of APC Antigen presentation DC-T cell assay Affinity of peptides and MHC MHC II
- 5. Importance of Immunogenicity in Therapeutic Biologics Development
- 6. Importance of Immunogenicity in Therapeutic Biologics Development
- 7. Effects on bio-availability Effect on Safety and Efficacy Effect on PK including potential cross reactivity to endogenous proteins Inhibition of the function of endogenous protein Injection site reactions Systemic reactions mild or life threatening Formation of ADA (HAMA, HACA, HAHA) Formation of neutralizing antibodies Formation of immune complexes Formation of anti-idiotypic antibodies EX. 1: first generation therapeutic mAbs EX. 2: terminate the development Impact of Immunogenicity
- 9. x Immoblize drug Add serum containing Anti-drug antibodies Detect with labeled (e.g. biotinylated drug) Immunogenicity Assessment Strategy - SIAT ® ADA Assay x x
- 10. Locate T cell epitopes 3D model of Fv domain (of the antibody) Multiple sequence alignmets (of human and murine) Tcell epitopes Replace T cell epitopes Affinity with HLA allotype Compatibility with the 3D structure Immunogenicity Assessment Strategy - SIAT ® De-immunization Service
- 11. Address: 45-1 Ramsey Road, Shirley, NY 11967, USA Tel: 1-631-871-5806 Fax: 1-631-207-8356 Email: info@creative-biolabs.com Web: www.creative-biolabs.com Contact us
Editor's Notes
- Hello, everyone. Welcome to watch the video named immunogenicity overview of therapeutic biologics which created by creative biolabs. In this video, we will introduce the definition of immunogenicity, background of immunogenicity and importance of immunogenicity therapeutic biologics development.
- What is immunogenicity? The concept of immunogenicity is more complex. In the video, Immunogenicity is defined as the propensity of the therapeutic biologics to generate immune responses to itself and to related proteins or to induce immunologically related non clinical affect or adverse clinical events. There are two types of immunogenicity in therapeutic biologics development process: wanted immunogenicity and unwanted immunogenicity Wanted immunogenicity is typically related with vaccines, where the injection of an antigen (the vaccine) stimulates an immune response against the pathogen (virus, bacteria, cancer cell...) aiming at protecting the organism. Unwanted immune responses to therapeutic biologics may also neutralize their biological activities and result in adverse events not only by inhibiting the efficacy of the therapeutic biologics, but also by cross-reacting to an endogenous protein counterpart, leading to loss of its physiological function (e.g., neutralizing antibodies to therapeutic erythropoietin cause pure red cell aplasia by also neutralizing the endogenous protein). The meaning of immunogenicity in this overview is the latter adverse immune response in the therapeutic biologics discovery and development.
- Antigen processing and presentation plays a critical role in immunogenicity of organisms. The processes are performed by professional APCs (antigen presentation cells) such as DCs, macrophages and B cells. There are two involved events in the process: 1. antigen capture that delivers antigens to the cellular antigen processing machinery; 2. antigen processing and presentation that generates antigenic peptides bound to MHC molecules for presentation to adaptive immune cells. The detailed process is illustrated here: 1. Extracellular antigens are captured by APCs through phagocytosis, macropinocytosis and receptor-mediated endocytosis. In acidic environment of endosome or lysosome, antigens are degraded into many immunogenic peptides which contain T cell epitopes. Following intake of antigens, MHC class II molecular is synthesized in the endoplasmic reticulum, and then transported by golgi apparatus to combine with antigen peptides to form peptide-MHC II complexes. 2. The complexes are presented to the surface of APCs. TCR can recognize the peptide-MHC II complexes to activate T cells to initiate an immunogenic response. The quality of the antigen presentation depends on affinity of the peptide-MHC complexes and there is a direct relationship between peptide-MHC complex stability and the immunogenic response. In addition, binding ability between APCs and CD4+ T cell is determined by DC-T cell assay.
- T cell-dependent and independent pathway involved in an immunogenic response. T cell-dependent pathway involves the uptake of biologics by antigen-presenting cells like immature dendritic cells (iDCs) and B cells. DCs process the biologic into peptides, mature and migrate to the T cell zone of the draining lymph nodes where they present the antigenic peptides to naive T cells expressing antigen-specific T cell receptors. This leads to T cell activation and proliferation. B cells can also take up biologic through their B cell receptor, process and present biologic-derived peptides to activated T cells that have migrated to the B cell zones. Activated T cells stimulate B cells resulting in the generation of antigen-specific antibody secreting plasma cells. T cell-independent pathway involves the direct stimulation of B cells by aggregated form of biologic. Marginal zone B cells can be stimulated by biologic bearing blood borne peripheral DCs. This pathway leads to generation of plasma cells that predominantly secrete IgM antibodies. Cross talk between these pathways contributes significantly towards the immunogenic response. ADA anti-drug antibodies, Ag antigen, bbDC – blood-borne peripheral dendritic cells, mDC mature dendritic cell, TCR T cell receptor, Th cell T helper cell, TLR Toll-like receptor.
- This is example roadmap for immunogenicity prediction. A potential step-wise approach to pre-clinical immunogenicity testing is depicted here. Step 1 would consist of the in silico screening of linear sequences from multiple therapeutic candidates for T cell epitopes and clusters. At the conclusion of Step 1, therapeutic candidates would be rated for immunogenic risk. Step 2 would be an in vitro evaluation of immunogenicity in a series of T cell assays. This phase could include examination of antigen processing and presentation, post-translational modifications, and different formulations. Subsequent Step 3 immunogenicity testing of therapeutic candidates may be carried out in vivo in established “humanized” animal models such as the Hu-SCID and HLA-transgenic mice. The culmination of this immunogenicity prediction strategy would be advancement into the drug development pipeline.
- Therapeutic biologics, including protein, enzyme, antibody, antibody-drug conjugate (ADC), are of significant value in the treatment of various diseases. “What about immunogenicity?” This is often the first question asked when a discussion about therapeutic biologics development occurs. Thus this issue has been the subject of a wide variety of reference material, and has been the subject of a cross-industry group that has written multiple white papers on aspects of immunogenicity. Immune responses to therapeutic biologics may pose problems for both therapeutic biologics efficacy and patient safety patient safety. Therefore, immunogenicity assessment is so important that it can not be ignored in the drug discovery process. Creative Biolabs launches siat immunogenicity system. Featured Services of SIAT System includes: siat insilico immunogenicity assessment, siat ex vivo immunogenicity assessment, siat in vivo immunogenicity assessment, siat anti-drug antibodies assays and siat de-immunogenicity.
- Effect of immunogenicity in the therapeutic biologics development can be summarize as follows:Effects on bio-availability, Effect on safety and efficacy, Effect on PK including potential cross reactivity to endogenous proteins, Inhibition of the function of endogenous protein, Injection site reactions, Systemic reactions mild or life threatening, Formation of ADA (HAMA, HACA, HAHA), Formation of neutralizing antibodies, Formation of immune complexes, Formation of anti-idiotypic antibodies. There are some real cases for illustrating the impact of immunogenicity. The first generation therapeutic mAbs were of murine origin, leading to highly adverse immune responses in patients because of the foreignness of the antibodies. Immunologically based adverse events, such as anaphylaxis, cytokine release syndrome, and cross-reactive neutralization of endogenous proteins mediating critical functions, have caused sponsors to terminate the development of what otherwise may have been efficacious therapeutic biologics. Fundamentally, with therapeutic biologics being developed today, the most important factor concerning immunogenicity is that it is a co-variate of pharmacokinetics, when immunogenicity against a therapeutic biologics occurs, it increases clearance and decreases exposure to that therapeutic.
- Characterisation and screening for physico-chemical determinants or formulation-based factors like impurities, heterogeneity, aggregate formation, oxidation and deamidation in the biologics will aid both in the prediction of immunogenicity and in the development of less immunogenic therapeutic agents. Moreover, predicting potential immunogenic epitopes in biologics will be an important and effective strategy to improve their safety and efficacy. A variety of preclinical immunogenicity screening strategies are being used during biologic development as listed here. SIAT immunogenicity assessment system from creative biolabs can achieve above goals.
- Arising of anti-drug antibodies (ADAs) against biotherapeutic drugs has been widely observed in clinical applications, such as anti-TNF adalimumab, 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. 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).
- Creative Biolabs offers SIAT® de-immunization service in conjunction with our SIAT® Immunogenicity Assessment. The workflow of our de-immunization service as is shown in the diagram. Step 1:Locate the T cell epitopes by our SIAT® immunogenicity assessment service. Step 2: Construct a 3D model of the Fv domain of the antibody. The model will help to study the location of predicted T cell epitopes and to evaluate the effects of a substitution on the stability and binding affinity of the antibody. The 3D structure is modeled by homology to the most related structure. Step 3:Make multiple sequence alignments of human and murine germline as a source of possible substitutions for T cell epitopes. Replace the T cell epitopes with a closest human germline sequence according to the multiple sequence alignments. If the possibilities of human germline are exhausted, continue with murine germline and then ad-hoc substitutions. All substitutions will be tested by the method in the next step. Step 4:Test the compatibility of the substitutions with the 3D structure of the antibody. Check the affinity of the substitutions with HLA allotypes. The overlapping peptides should not be transformed into T cell epitopes. The substitutions should avoid inducing strain in the antibody scaffold. If all tests are passed, accept the substitution. Otherwise, new substitutions are evaluated and selected.
- If you are interested in learning more about our siat immunogenicity assessment system, please don’t hesitate to contact us at info@creative-biolabs.com or call us at 1-631-871-5806. We will be more than happy to serve you!