Immunogenicity Prediction and Control

Regulatory Perspectives, Risk Factors, and Management

October 5 - 6, 2022 ALL TIMES EDT

The impact of immunogenicity on safety and efficacy – and consequent cost to the industry – is well understood. Accordingly, investigators are focusing on factors that contribute to immunogenicity, as well as a number of different approaches to predict immunogenicity at the drug discovery stage. There are several major problematic areas with novel modalities. Efforts are being made to suppress immune responses to these products and to introduce tolerizing and deimmunization approaches. Attend in 2022 to hear insightful feedback from the FDA and learn from top industry and academics in this field.

Wednesday, October 5

Registration Open (Arlington Registration)12:00 pm


ROOM LOCATION: Arlington Salon V

12:45 pm

Chairperson's Opening Remarks

Michael Partridge, PhD, Director, Bioanalytical Sciences, Regeneron


Differential Immune Responses to Deamidated Adeno-Associated Virus Vector

Ronit Mazor, PhD, Principal Investigator, CBER, FDA

Adeno-associated viruses (AAV) are potent vectors used for gene therapy, but recent clinical findings have revealed immunogenicity-related challenges, including formation of a cytotoxic immune response against AAV capsid protein in transfected cells. We report that deamidation, a spontaneous post-translational modification that occurs in multiple AAV serotypes, can result in formation of CD4 T cell epitopes with a Th1 cytokine pattern in donor samples with specific HLA backgrounds.

1:20 pm

Immunogenicity of AAV-Based Gene Therapy

Klaudia Kuranda, PhD, Head of Immunology, Spark Therapeutics, Inc.

Adeno-associated virus (AAV) gene transfer is a promising treatment for many patients with life-threatening genetic diseases. However, host immune response to the vector seems to pose a challenge for the durability and safety of AAV-mediated gene therapy. Today, AAV immunogenicity impacts on trial design and patient eligibility and remains a focus of preclinical studies that attempt to minimize host immune response against therapeutic vectors.

1:50 pm

Nonclinical Pharmacodynamic Effects and Immunogenicity Assessment of Prophylactic Immune Modulation Prior to Gene Therapy Dose Administration in C57BL/6 Mice

Brian Long, PhD, Principal Scientist, Clinical Immunology, BioMarin Pharmaceutical, Inc.

Clinical programs investigating AAV-vectored gene therapies have adopted the use of immune modulatory therapies to prevent or treat inflammatory responses and preserve transgene expression. We evaluated the pharmacodynamic and immunomodulatory effects of seven different immunosuppressive agents administered prophylactically prior to gene therapy dose administration and continued for four weeks in C57BL/6 mice. This study demonstrates prophylactic immune modulation prior to AAV administration can improve liver-directed transgene expression in mice.

2:20 pm An Integrated Approach to Managing Immunogenicity Risk and Optimum Protein Design

Emilee Knowlton, Senior Immunology Sales Specialist, Sales, ProImmune, Inc.

Immunogenicity risk assessment is an essential step in bringing therapeutic drugs to the market. ProImmune's risk management tools evaluate immunogenic epitopes and the corresponding functional T cell responses that can lead to unwanted immune responses.  Case studies will highlight how the integrated platform is used to address key questions in the drug development phase. 

Refreshment Break in the Exhibit Hall with Poster Viewing (Arlington Salon IV)2:50 pm


Understanding and Navigating Immune Responses to Proteins Used as Therapeutics and in Novel Modalities Like Gene Editing

Zuben Sauna, PhD, Research Biologist, CBER, FDA

Immunogenicity refers to immune responses to proteins used in therapeutic applications. Immunogenicity is of concern during drug development and licensure as it can affect the safety and/or efficacy of drug products. The presentation will provide: (1) An overview of tools available for immunogenicity risk assessment during drug development and licensure. (2) Specific examples that illustrate the use of in silico, in vitro, and ex vivo methods for immunogenicity risk assessment of proteins used therapeutically and those used in emerging technologies such as gene editing. Judicious application of tools available for immunogenicity risk assessment can permit better decision-making and potentially safer drugs.

4:00 pm

A Case Study of Unpredicted Clinical Immunogenicity of a Heteroimmunoglobulin

Troy Barger, Senior Scientist, Amgen

Evolving drug modalities present ever-increasing challenges to immunogenicity prediction and risk assessment. In this case study, a heteroimmunoglobulin antibody against TL1A and TNF alpha was observed to have a high rate of immunogenicity in a clinical study contrary to risk assessment from available predictive tools. Characterization of the immune response is described focusing on the bioanalytical data. Finally, formation of large complexes is proposed to account for the immunogenicity observed.

ROOM LOCATION: Arlington Salon V


Immunogenicity Assessment of Gene Therapy Products


Michael Partridge, PhD, Director, Bioanalytical Sciences, Regeneron

  • Assessment of pre-existing antibodies to gene therapy vectors and the potential impact of these antibodies on the therapy
  • Discussion around T cells: An important issue for many gene therapy products
  • Assays: Is the protein therapeutics paradigm applicable? Tiered assessment; Binding ADA and NAb (inhibitor); Conservative 5%/1% false positive concept; Cut point determination. E.g. Hemophilia GTx Guidance and recommendations for assessment of inhibitors. Is assessment of binding antibodies recommended as well?
  • Are these elements needed for assessment of immunogenicity to the components of the GTx and the transgene?
  • Is an integrated summary of immunogenicity recommended? 
  • Is an immunogenicity risk assessment recommended?

Andrew Byrnes, PhD, Chief, Gene Transfer and Immunogenicity Branch, CBER, FDA

Brian Long, PhD, Principal Scientist, Clinical Immunology, BioMarin Pharmaceutical, Inc.

Klaudia Kuranda, PhD, Head of Immunology, Spark Therapeutics, Inc.

Close of Day5:00 pm

Thursday, October 6

Registration and Morning Coffee ( Arlington Foyer)7:30 am


ROOM LOCATION: Arlington Salon V

7:55 am

Chairperson's Remarks

Jack A. Ragheb, MD, PhD, Senior Vice President Translational Sciences and Medicine, NexImmune

8:00 am

Adaptive Responses to SARS-CoV-2 Infection and Vaccination

Daniela Weiskopf, PhD, Research Assistant Professor, La Jolla Institute for Immunology

I will present data from several studies performed in the last two years. The results show that SARS-CoV-2 infection induces multi-specific and multi-functional adaptive responses which is durable over the 6-8 months period. Neutralizing antibodies are likely key to protect from infection, but substantial contributions of T cells likely at the level of protection from severe disease. Side-by-side longitudinal comparison of different vaccine platforms reveals durable B and T cell responses, and allow correlation with vaccine efficacy. T cell reactivity is largely preserved at the level of SARS-CoV-2 variants, including Omicron and Delta.



Understanding and Circumventing the Immune Responses to Approved Protein Therapeutics

Daniel LaGasse, PhD, Research Regulator, CBER, FDA

Immunogenicity can compromise the safety and/or efficacy of therapeutic protein products. Due to the serious conditions they treat and lack of alternatives many protein therapeutics are approved even if they elicit immune responses. In the clinic, in addition to poor patient outcomes, the social and economic costs associated with neutralizing antibodies are considerable. In this presentation, I survey the immunogenicity of approved therapeutic proteins, discuss strategies for clinical management of immunogenicity, and identify challenges associated with circumventing the immune responses to approved protein therapeutics.

9:00 am Development of an iLite Reporter Cell Platform for the Quantification of Anti-AAV Neutralizing Antibodies

Jordi Rodó Morera, PhD, R&D, Svar Life science

The efficacy of gene therapies using recombinant AAV can be limited by an antibody-mediated immunity against the virus. AAV infections, potentially occurring in early childhood, may result in a pre-existing immunity to one or more AAV serotypes. Here we present a novel two-component iLite reporter cell system for the detection and quantification of neutralizing antibodies against AAV vectors. This assay allows for reliable assessment of anti-AAV antibodies in clinical applications.  

Breakout Discussions9:30 am

Breakout discussions provide an opportunity to discuss a focused topic with peers from around the world in an open, collegial setting. Select from the list of topics available and join the moderated discussion to share ideas, gain insights, establish collaborations or commiserate about persistent challenges. Please visit the breakout discussions page on the conference website for a complete listing of topics and descriptions.

BREAKOUT DISCUSSION: Risk Assessment and Mitigation Strategies for Immunogenicity of Life Saving Therapeutic Proteins and Novel Treatment Modalities: Deimmunization and Immune Tolerance Strategies

Amy Rosenberg, MD, Senior Director of Immunology and Protein Therapeutics, Epivax

  • Strategies to prevent or mitigate immunogenicity of therapeutic proteins and application of these strategies to prevent/mitigate immunogenicity of novel modalities: Cell and gene therapies​
  • - For what clinical scenarios are each strategy (deimmunization, immune suppression, therapeutic drug monitoring, immune tolerance induction) appropriate
    - Retreatment of patients with AAV vectored therapeutics is blocked by humoral immunity to capsid
    - Preexisting immunity to key elements of gene therapies (AAV, Cas9 proteins) may preclude patients from entry into clinical trials
  • Transplantation of gene-corrected Autologous Hematopoietic Stem Cells mediated by lentiviral vectors in patients with “inborn errors of metabolism”- strategies for implementation
    -  Can lentivirus integration be targeted to avoid integration into oncogenes leading to tumorigenicity? To optimize expression/production of the therapeutic
    - What level and duration of immune suppression will be required post-transplant to prevent rejection of gene-corrected Autologous HSC?

Immunogenicity Prediction – Translation into the Clinic

Klaudia Kuranda, PhD, Head of Immunology, Spark Therapeutics, Inc.

  • Are animal models of gene transfer able to predict immune response in humans? While immune-related toxicities seem to be rare in the clinic, is there a possibility that animal models are predictive, but the number of animals used for the preclinical studies is too low? 
  • The immunogenicity assay toolbox improves every day. Do we have the right assays today to detect anti-AAV or anti-transgene responses in animals? Can in vitro assays be used instead and which one?
  • How can the preclinical data generated be used in the clinic?  What is the true value of the pre-clinical assays and how can this be proved?​

BREAKOUT DISCUSSION: The Complex Landscape of the MHC-II Ligandome

Laura Santambrogio, PhD, Professor, Associate Director, Precision Immunology, Weill Cornell Medicine

  • The multiplicity of translational and post-translational mechanisms affecting the MHC-II restricted immunopeptidome   
  •  How to quantitatively assess the MHC-II ligandome and why it's important to know epitope copy numbers  
  •  MHC-II presented tolerogenic and immunogenic peptides  ​

Coffee Break in the Exhibit Hall with Poster Viewing (Arlington Salon IV)10:20 am



Tolerance and Check Point Inhibitors

Jack A. Ragheb, MD, PhD, Senior Vice President Translational Sciences and Medicine, NexImmune

11:20 am

Tolerogenic Treatment of Autoreactive T and Antibody Responses in a Mouse Model of Neuromyelitis Optica Using Aquaporin 4-Encapsulating PLG Nanoparticles

Stephen Miller, PhD, Professor of Microbiology-Immunology, Feinberg School of Medicine, Northwestern University

Neuromyelitis optica (NMO) is an autoimmune demyelinating disease of the CNS characterized by T cell and Ab responses to the water channel protein Aquaporin 4 (AQP4). We developed a robust NMO model in C57BL/6 mice characterized by AQP4-specific Th17 and Ab responses resulting in high disease incidence and severity. We demonstrate the ability of tolerance induced by AQP4-encapsulating PLGA nanoparticles to both prevent and treat ongoing disease.

11:40 am

Protein Resurfacing Reduces Binding by Preexisting Anti-Drug Antibodies and Alleviates Hypersensitivity Responses

Daniel Leventhal, PhD, Head of Immunogenicity, Generate Biomedicines

Anti-drug antibodies (ADAs) and hypersensitivity responses limit the use of many biotherapeutics. Generate Biomedicines has developed a generalizable method for computationally designing resurfaced proteins to avoid binding by preexisting ADAs. This technique was applied to L-asparaginase (ASN) to produce functional variants with up to 58 surface mutations. Resurfaced ASNs exhibited reduced binding to mouse, rabbit, and human ADAs, and alleviated hypersensitivity in an in vivo mouse model.



Innate Immune Response Modulating Impurities Testing for Biosimilars: Where We Are and What We Are Missing

Daniela Verthelyi, MD, PhD, Chief, Laboratory of Immunology, CDER, FDA

Comparative in vitro analytical methods to characterize innate immune response modulating impurities could potentially provide a more robust understanding of immunogenicity risk for biosimilars and help streamline their development. This talk will discuss the risks posed by innate immune response modulating impurities, available assays, and data interpretation, as well as common pitfalls and remaining knowledge gaps.

12:30 pm Biotherapeutic Internalization as an Immunogencity Risk Assessment Tool

Lynn Kamen, PhD, Scientific Officer & Executive Director, BioAgilytix

As therapeutic antibodies evolve, the potential for anti-drug antibody (ADA) responses become more likely, making in vitro immunogenicity prediction assays an important tool for drug development.  A key component of the immune response is the uptake of antibody by antigen presenting cells (APCs).  This presentation will highlight the development of a dendritic cell (DC) internalization assay that measures antibody internalization and shows a correlation between the level of internalization and immunogenicity.

1:00 pm LUNCHEON PRESENTATION:A Strategy for Building a Powerful Immunosafety Risk Assessment to Support Regulatory Submissions

Noel Smith, Head of Immunology, Early Development Services, Lonza

An understanding of the potential immunogenicity and immunotoxicity risk of your drug candidate is a key part of pre-clinical development. Human primary immune cell assays can provide crucial information on both the innate and adaptive immune response induced by a drug candidate. We discuss how these assays can be optimized and qualified to ensure the data is highly sensitive, accurate and robust and can effectively support both lead selection and regulatory filings. 

Session Break1:30 pm


2:00 pm

Chairperson's Remarks

Laurent P. Malherbe, PhD, Research Advisor, Eli Lilly & Co.

2:05 pm

Predictive Assays, Studies, and Tools: How Can These Be Improved?

Marc Pallardy, PhD, Professor & Team Leader, Inserm UMR 996, University of Paris-Saclay

The presence of protein aggregates in therapeutic products has been correlated with ADA production and it has been shown that aggregates could play the role of danger signals facilitating T cell-dependent immune responses. Aggregates could be recognized by professional antigen-presenting cells, such as dendritic cells (DC), inducing their maturation and the activation of CD4+ T cells and thus helping B cell maturation and production of high-affinity ADA.

2:25 pm

Towards Harmonization of in vitro T Cell Assays to Predict Immunogenicity of Biologics

Laurent P. Malherbe, PhD, Research Advisor, Eli Lilly & Co.

In vitro immunogenicity tools are used during the development of biologics to select clones with decreased clinical immunogenicity risk. Here we will discuss ongoing efforts to benchmark in vitro T cell activation assays and the cross-industry work led by HESI and AAPS to develop common reference biologics, a critical first step toward assay harmonization.

2:45 pm

CANCELLED: Immunogenic and Tolerogenic Peptides

Laura Santambrogio, PhD, Professor, Associate Director, Precision Immunology, Weill Cornell Medicine

I will discuss all the molecular components that regulate the selection of the MHC-II presented peptides and the variables which contribute qualitatively and quantitatively, to the composition of the MHC II ligandome. Altogether, ensuring that the immunopeptidome landscape is highly sensitive to any changes in the composition of the intra- and extracellular proteome for a comprehensive survey of the microenvironment for MHC II presentation to CD4 T cells.


3:05 pm

Integrated Approach to Assess Immunogenicity Risk of Biotherapeutic Products

Sivan Cohen, PhD, Senior Principal Scientist, Genentech

Biotherapeutics have revolutionized the treatment of many diseases. However, unwanted immune responses still present a major challenge to their widespread adoption. Many patients treated with biotherapeutics develop anti-drug antibodies (ADAs) that may reduce the efficacy of the therapy or cross-react with the endogenous counterpart of a protein therapeutic, or both. Therefore, predicting the risk for immunogenicity of biotherapeutic products at early stages is a crucial need. This presentation will focus on in silico and in vitro T cell assay studies to characterize the immunogenic potential of different biotherapeutic proteins and their correlation to the clinically observed outcome.

3:25 pm

Assessing Immunogenicity Using Immune Humanized Mice

Kristina E. Howard, Principal Investigator, Division of Applied Regulatory Science (DARS), CDER, FDA

Close of Conference3:45 pm