Part Two: Immunogenicity Prediction and Control - Immunogenicity Summit





8:30am Chairperson’s Remarks

Matthew Baker, Ph.D., Chief Scientific Officer, R&D, Antitope Ltd.

8:35 High Pressure Dissociation of Aggregate Structure and Resulting Decreased Immunogenicity

Matthew Seefeldt, Ph.D., Vice President, Research, BaroFold, Inc.

Studies have identified the causal relationship of the presence of small levels of aggregates in protein-based therapeutics to the development of immunogenicity toward native proteins after repeated dose administration. BaroFold’s novel, proprietary, high pressure refolding technology is effective for the reversal of aggregates present in commercial drug formulations and is scaleable in the GMP setting. We have demonstrated proof-of-principle on commercial growth hormone formulations, decreasing the immunogenicity of Nordiflex and IFN-beta-1b formulations in murine models. This talk will summarize the lessons that we have learned in the context of aggregate size, structure, content and immunogenicity.

9:05 Impact of Product Related Factors (CMC factors) on Immunogenicity

Satish SinghSatish K. Singh, Ph.D., Research Fellow, Biotherapeutics Pharmaceutical Sciences, Pfizer, Inc.

Several factors influence the generation of an immune response including patient characteristics, disease state and the therapy itself. Product-related factors such as the molecule design, the expression system, post translational modifications, impurities, contaminants, formulation and excipients, container/closure as well as degradation products are also implicated. The talk will critically examine the available data for the impact of these latter factors on clinical immunogenicity. Deconvolution of the clinical impact of product attributes from patient susceptibility is not readily feasible.

Bernard Malliere9:35 Quantification of Pre-Existing CD4 T Lymphocytes Specific to Therapeutic Proteins as a Tool of Evaluation of Immunogenicity Potential

Bernard Maillere, Ph.D., Head, Immunochemistry, Institute of Biology and Technologies, CEA

We have quantified the number of pre-existing CD4 T lymphocytes specific to murine, chimeric, humanized and human therapeutic antibodies in the blood of healthy individuals. Irrespective of the humanization levels, the number of antibody-specific CD4 T lymphocytes was in good concordance with their known immunogenicity. We also successfully applied this approach to human erythropoietin and provide a possible explanation for the immune response that was observed for certain batches of recombinant EPO.

10:05 Predicting and Assessing the Immunogenicity of Adnectins, a Novel, Proprietary Class of Targeted Biologics

Nick Marsh, Ph.D., Director, Pharmacology, Adnexus Therapeutics, Inc.

10:35 Networking Coffee Break, Poster and Exhibit Viewing



11:10 Immune Tolerance Mechanisms: Animal Models and Approaches

David W. Scott, Ph.D., Vice Chair, Research, Uniformed Services, University of Health Sciences, Bethesda

This presentation will focus on the parameters for immunogenicity of biotherapeutics and how an understanding of tolerance mechanisms can be used to control undesirable immune responses. The nature of the multiple factors impacting on immunogenicity and approaches to control them will be discussed.  Novel and classical approaches to induce tolerance will be reviewed together with their advantages and disadvantages. The uses of IgG chimeric fusion proteins and gene therapy, for example, have provided additional insights for tolerance mechanisms.  Moreover, this has led to a focus on regulatory T cells in this and other models.  These studies have an impact on the design of protein therapeutics for reduced immunogenicity and adverse events.  

11:40  The Absence of the G1m1 Allotype may be a Risk Factor for Anti-antibody Immunogenicity

Fiona Harding, Ph.D., Senior Principal Research Scientist, Abbott Biotherapeutics


12:10 pm Sponsored Presentation (Opportunity available, please contact Ilana Quigley,

12:25 Luncheon Presentation or Lunch on Your Own (Opportunity available, please contact Ilana Quigley, 


2:00 Chairperson’s Remarks

David W. Scott, Ph.D. Vice Chair for Research, Uniformed Services University of Health Sciences, Bethesda, MD

2:05 A Human Lymphoid Organ Model (HuALN) For Predictive Testing of Immunogenicity, Immunotoxicity and Immune Functions in vitro

Christoph GieseChristoph Giese, Ph.D., Director, Cell and Tissue Services, ProBioGen AG

Human tissue based models which emulate immune organ function are conceived to bridge the gap in testing immune functionality, immunotoxicity and predictive immunogenicity between early lead optimization and the pre-clinical development stage. The model of the Human Artificial Lymph Node (HuALN) is designed to investigate induced immune responses in vitro. The 3D organoid model can be used for long-term culture and repeated dosing. Cytokine release, antibody secretion, cellular functionality and tissue formation are monitored.

2:35 In Silico Prediction of Immunogenicity: Sense and Non-Sense

Philippe Stas, M.B.A., Head, Applied Protein Services, Lonza Biologics; Head, Algonomics NV

Different methods and algorithms exist for the prediction of CD4+ T-cell epitopes. As these epitopes are a requirement for a sustained immunogenicity, selection of protein therapeutics based on their relative epitope content has been applied for over 2 decades. This presentation focuses on the use of in silico methods for lead selection, and how they complement the in vitro strategies for immunogenicity assessment. In specific cases, antibody engineering techniques can be used to reduce or avoid immunogenicity of the drug. Selected case studies are presented to explore the clinical relevance of predictive methods in biotherapeutic development.

Frank Carr3:05 Generation of Therapeutic Proteins Against Inflammatory Disease and Cancer Targets with a Low Risk of Clinical Immunogenicity

Frank J. Carr, Ph.D., Director, Biologics Research, Antitope Ltd.

CD4+ T cell epitopes in the variable region sequences of therapeutic proteins have been identified by ex vivo T cell epitope mapping. The number and potency of T cell epitopes correlated with the immunogenicity (frequency of anti-therapeutic responses) of these proteins in the clinic. Furthermore, the presence of aggregated antibody induced more frequent ex vivo T cell responses. Data will be presented that provides evidence linking the presence of T cell epitopes in the sequences of therapeutic proteins with immunogenicity observed in patients as well as how proteins can be engineered to avoid T cell epitopes.

3:35 Networking Refreshment Break, Poster and Exhibit Viewing

4:00 Reducing the Immunogenicity of the Pseudomonas Exotoxin-Based Immunotoxins by Modification of B Cell Epitopes

Masanori OndaMasanori Onda, M.D., Ph.D., Laboratory of Molecular Biology, NCI/NIH

Our approach to de-immunize our immunotoxin is to identify the B cell epitopes and to modify them by mutagenesis. The mutated new immunotoxin was fully active and of extremely low immunogenicity. This approach can also be applicable for other protein therapeutics.

4:30 Design Space Available for Non-immunogenic Proteins

Stephen Lee, Ph.D., Associate Professor, Biomedical Engineering, Ohio State University

My group has derived an algorithm which explicitly identifies literally all possible variants of any protein that lack binding motifs for preselected class I or class II haplotypes, based on set theory. It is useful in vaccine or non-immunogenic protein design. The number of possible epitope-free variants can be immense (>10^10 in many cases). Along with data from deep protein mutagenesis experiments, this suggests identification of non-immunogenic protein sequences should be possible today, though discovery of the subset of biofunctional variants will involve laborious, deep mutagenesis, and non immunogenic proteins may be very different in sequence from their parents.

5:00 End of Part Two

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