Cambridge Healthtech Institute’s 11th Annual

Immunogenicity Assessment & Clinical Relevance  

Assay Strategy for Meaningful Evaluation

October 16-17, 2019

 

The industry continues to be challenged by the development, application and interpretation of immunogenicity assays. The FDA has advocated a more stringent approach for cut point setting and assay validation creating further difficulties. Moreover, the industry remains uncertain about when the more challenging neutralizing antibody assays should be applied and which type of assay is reliable and acceptable. Additional ongoing challenges concern managing drug and target interference, understanding and handling the impact of pre-existing antibodies, and interpreting the clinical significance of assay data.  

Final Agenda

Wednesday, October 16

7:30 am Registration and Morning Coffee

ASSESSING THE CLINICAL RELEVANCE OF ASSAYS

8:25 Chairperson’s Opening Remarks

Sandra Garces, MD, PhD, Clinical Research Director, Global Drug Development, Amgen


8:30 KEYNOTE PRESENTATION: Designing a Therapeutic Drug Monitoring Program to Optimize the Management of Biologic Therapies in Routine Clinical Practice

Sandra Garces, MD, PhD, Clinical Research Director, Global Drug Development, Amgen

In clinical practice, significant heterogeneity is observed in therapeutic responses to biologic therapies, varying from non-response to loss of response or to complete clinical remission. Therapeutic drug monitoring (drug levels and ADA) can work as a tool to help us understand clinical heterogeneity and to adopt tailored therapeutic decisions that can improve the cost-effectiveness of biologic therapies. Different types of algorithms can be designed according to the specific goals and type of assays available. Practical cases will be used to better illustrate when and how physicians can use such information to guide therapeutic decisions in clinical practice, highlighting the impact different PK and ADA assays may have in the interpretation of the results.

9:00 An Integrated Approach to Assessing the Presence of Neutralizing Activity for Biopharmaceuticals

Wu_YulingYuling Wu, PhD, Associate Director, Clinical Pharmacology Biologics and Bioanalysis, AstraZeneca

Multiple bioanalytical assays, including screening/confirmatory assays, NAb (neutralizing antibody), PK (pharmacokinetic), and PD (pharmacodynamic) assays, are commonly developed and applied to different stages of clinical development of biopharmaceuticals. The industry remains uncertain about when the more challenging NAb assays would be applied and how to effectively use PK/PD/ADA integrated data to better understand the presence of neutralizing activity.

9:30 KEYNOTE PRESENTATION: FDA Regulatory Perspective on Clinical Relevance of Immunogenicity Assay Validation

Kong_LeopoldLeopold Kong, PhD, Biotech Quality and Immunogenicity Reviewer, FDA 

Anti-drug antibody (ADA) assay should be sufficiently sensitive to detect ADA before they reach levels associated with altered pharmacokinetic, pharmacodynamic, safety, or efficacy profiles. We will discuss key points and considerations in determining assay suitability in the context of product immunogenicity risk and clinical experience. We will highlight a case study in which analysis of available clinical pharmacology data in addition to validation data was necessary to assess assay suitability.

10:00 Coffee Break in the Exhibit Hall with Poster Viewing

DEVELOPMENT OF NEUTRALIZING ANTIBODY ASSAYS

10:40 NEW: Advantage of Cell-based Binding Assay for Neutralization Antibody Assay

Tatyana Yun, PhD, Senior Scientist, Merck

For most of the mAb biotherapeutics targeting cell surface protein, a competitive ligand binding assay is possible for neutralizing antibody testing. However, recombinant protein may not fully reflect natural counterpart and may introduce artificial effects. Here we share a couple of case studies where we utilize the Meso Scale Discovery platform to develop cell-based binding assays, which is easier and generally more sensitive than cell-based functional assays, while providing more physiological binding than competitive LBA.

11:10 FEATURED PRESENTATION: Highly Drug Tolerant Neutralizing Antibody (NAb) Assays: New Approaches to Drug Removal in Competitive Ligand Binding Assays

Michael Partridge, PhD, Associate Director, Bioanalytical Sciences, Regeneron Pharmaceuticals

Neutralizing antibody (NAb) assays typically have poor drug tolerance (DT), potentially underestimating NAb incidence. This presentation will describe new approaches to improve DT using bead-based drug depletion methods that avoid interference due to carryover of the drug-capture reagent. Analysis of ADA positive clinical samples with elevated drug levels using these techniques detected more NAbs. However, these NAb positive samples were low titer and did not appear clinically relevant.

SVAR 11:40 Development of an ILite Reporter Cell Lines for the Quantification of Anti-AAV Neutralizing Antibodies

Tovey-MichaelMichael Tovey, Managing Director, Paris Management, Svar Life Science X

The efficacy of adeno-associated virus (AAV) mediated gene-therapy is limited by the development of antibodies against the virus capsid. A cell-based assay for the quantification of anti-AAV neutralizing antibodies (NAbs) will be described that reflects the effect of anti-AAV NAbs on the expression of recombinant AAV vectors.


11:55 Development of a Neutralizing Antibody Assay for Kadcyla

Chung_ShanShan Chung, PhD, Associate Director and Principal Scientist, Genentech, Inc.

Kadcyla is an antibody drug conjugate composed of a humanized anti-HER2 antibody (trastuzumab) covalently conjugated to a cytotoxic drug (DM1). Kadcyla is indicated for the treatment of patients with HER2-positive metastatic breast cancer. This presentation will describe in vitro characterization of Kadcyla for biological activities pertinent to its mechanisms of actions, and development of a fit-for-purpose neutralizing antibody assay to support characterization of Kadcyla-specific anti-drug antibodies in human serum.

12:25 pm Lunch on Your Own

MANAGING DRUG INTERFERENCE AND DRUG TOLERANCE

1:40 Chairperson’s Remarks

Sandra Garces, MD, PhD, Clinical Research Director, Global Drug Development, Amgen

1:45 Assay Strategies to Monitor Immunogenicity of New Antibody Therapeutics in Ophthalmology

Stubenrauch_KayKay Stubenrauch, PhD, Head of Large Molecule Bioanalytical Sciences 1, Roche Diagnostics

New antibody therapeutics are increasingly used to treat chronic ocular diseases. Corresponding bioanalysis (PK, ADA, PD) is challenging with respect to limited ocular sample availability and accessibility. Improved approaches, alternative assay formats, and new technologies are required – for instance, to adequately consider the drug influence on ADA and soluble target analysis in Ophthalmology.

2:05 Impact of Pre-existing Antibodies on Cut Point Determinations and Immunogenicity Assessment Strategy

Niu_HongmeiHongmei Niu, PhD, Senior Scientist, EMD Serono

Pre-existing antibodies are frequently observed in clinical settings. Irrespective of the clinical outcomes, the prevalence of pre-existing antibodies at baseline imposes an analytical burden. A few case studies will be presented to describe mitigation approaches on assessment of an appropriate assay cutpoint and detection of the treatment-emergent ADA response in the presence of the pre-existing antibodies.

 

2:25 Approaches to Resolve False Reporting in Neutralizing Antibody Assays Caused by Reagent Leaching from Affinity Capture Elution Solid Phase

Xiang_YuhongYuhong Xiang, Senior Scientist and Manager, Jounce Therapeutics

Insufficient drug tolerance presents a major challenge in neutralizing antibody (NAb) assays development for biotherapeutics. A case study of SPEAD in conjunction with a competitive ligand binding NAb assay is presented. A significant degree of biotin-drug conjugate leaching was observed resulting in false reports in NAb assay. Mitigation approaches have been evaluated to address drug/biotin drug conjugate leaching. Optimized conditions enabled a significant reduction of drug/biotin-drug conjugate leaching.

2:45 Problem Solving Round Table Discussions

Please join us for a moderated round table discussion with brainstorming and interactive problem solving. These sessions bring together attendees from diverse backgrounds to exchange ideas and develop future collaborations around a focused topic, in an informal environment.

TABLE 1: Recent Advances with Novel Modalities: Gene Therapy, CAR-T therapy, Peptides and More

Moderator: Jim McNally, PhD, Executive Director, Head of Research and Clinical Assays, Program Lead, CRISPR Therapeutics

  • Recent data on pre-existing reactivity for AAV
  • Immunogenicity assessment of novel modalities
  • Application of current guidance to novel modalities
  • What is your product? The vector, the expressed product?

TABLE 2: Strategies to Improve Drug Tolerance and Manage Interference

Moderator: Lilia Macovei, PhD, Bioanalytical Principal Investigator, Senior Scientist, Pfizer

  • Dissecting strategies for managing the drug/target interferences and drug tolerance in Gene therapy programs vs LBA
  • Case studies of matrix interference in ADAs and Nabs

TABLE 3: Individual vs. Population-Risk Assessment of Immunogenicity – Relevance for Labeling Information

Moderator: Sandra Garces, MD, PhD, Clinical Research Director, Global Drug Development, Amgen

  • Differences between the two
  • Assessing the risk of IMG at a population-level and defining the impact of IMG on the overall benefit-risk profile of the product
  • How to demonstrate a causal association between the development of ADA and undesirable clinical outcomes and assess the impact and clinical relevance of those outcomes in the specific patient population
  • How to define clinically useful information on IMG for labelling purposes

TABLE 4: Assessing Immunogenicity in Oncology Trials

Moderator: Mohamed Hassanein, PhD, Staff Scientist, Bioanalytical Sciences, Regeneron Pharmaceuticals

  • Immunogenicity assessment Immune checkpoints blockers mAbs
  • Challenges and solutions for evaluating NAbs of bispecifics and multi domain biologics
  • Immunogenicity assessment for IO new therapeutic modalities (i.e. Oncolytic viruses, therapeutic vaccines, DNA editing vectors)
  • Immunogenicity assessment for cell-based therapeutics (i.e. CAR-T, TILs)

NEW TABLE 5: The Challenge of Drug- and Matrix-Interference in Immunogenicity Testing

Moderator: Tatyana Yun, PhD, Senior Scientist, Merck

  • Current practice to overcome drug and matrix interference
  • Case studies where desired drug tolerance cannot be achieved
  • Issues with current practice of overcoming drug and matrix interference

NEW TABLE 6: Assay Development and Validation: Selecting the Optimal Format for your Assay, Overcoming Technical Challenges, Data Reporting and Cut Point Calculations

Moderator: Michael Partridge, PhD, Associate Director, Bioanalytical Sciences, Regeneron Pharmaceuticals

 

NEW TABLE 7: What Does the FDA Want to Know? Case Studies on Immunogenicity Related to Generic Peptides and Biologics

Annie De Groot, MD, CEO/CSO, EpiVax

  • In silico
  • In vitro
  • MAPPS

3:40 Refreshment Break in the Exhibit Hall with Poster Viewing

RECENT ADVANCES IN GENE THERAPY

4:20 Importance of Pre-existing Abs to the Viral Capsid during Immunogenicity Assessment of Viral Vectors based Gene Therapy

Jim_McNallyJim McNally, PhD, Executive Director, Head of Research and Clinical Assays, Program Lead, CRISPR Therapeutics

This talk will focus on the assessment of pre-existing antibodies to the viral capsid and the implications for their impact on the successful dosing of gene therapy drug candidates. Case studies will show how the thought process about pre-existing antibodies is evolving. There will be a focus on the tools used to measure pre-existing antibodies against the viral capsid and their utilization as exclusion criteria for entry into clinical studies.

4:40 Bioanalytical Support for Gene Therapy Programs: Strategies and Hurdles

MacoveiLilia Macovei, PhD, Bioanalytical Principal Investigator, Senior Scientist, Pfizer Inc.

Recombinant viral vectors have been reliable tools for basic research for many years and are now making their way into translational use with the use of rAAVs in early clinical trials as delivery platforms. Their tissue tropism and relative safety due to their low rate of genomic integration represent key features making rAAVs promising instruments as vectors for future gene therapy drugs. In this presentation, we will discuss the bioanalytical assay development for the adeno-associated viral vectors used for an AAV-based investigational product. The anti-capsid and anti-transgene ADA assay development, unique challenges, and immunogenicity strategy to detect anti-capsid neutralizing antibodies will be presented.

5:00 Immunogenicity Assessment of AAV Gene Therapy Vectors Combined with Tolerogenic Particles

SLeung2Sheldon Leung, PhD, Director, Bioanalytical Laboratory, Selecta Biosciences

As with all biologics, a thorough immunogenicity assessment of AAV is important for safety and efficacy. In addition to the evaluation of anti-capsid and neutralizing antibodies, the assessment of AAV gene therapeutics can include detection of anti-transgene antibodies as well as cell-mediated and innate immune responses. Further characterization of isotypes and binding epitopes can provide additional useful information. Pre-clinical immunogenicity data within the context of ImmTOR technology (tolerogenic particles) will be discussed.

5:20 Considerations and Strategy for Detection of Pre-Existing Neutralizing Antibody in AAV Gene Therapy

JiangZhihua Jiang, PhD, Principal Scientist, BioMedicine Design, Pfizer Inc.

Pre-existing neutralizing antibodies (NAb) against AAV capsids interfere with the efficacy of AAV-based gene therapy, even at low antibody titers. Screening and titering anti-AAV NAb in enrollment of subjects is critical for therapeutic success in both nonclinical and clinical studies. Though cell-based assay is widely used for anti-AAV NAb titration, insufficient assay sensitivity reduces prediction value of NAb titer on therapeutic efficacy. Additionally, lack of standardization of detection methods causes the difficulties in comparing the prevalence of NAb positivity among different populations and the thresholds of NAb titer for subjects enrolled in various studies. This presentation will discuss the detection methods for anti-AAV NAb, the strategies in improving assay sensitivity, and the standardization of method.

5:50 Welcome Reception in the Exhibit Hall with Poster Viewing

6:50 Close of Day

Thursday, October 17

7:30 am Morning Coffee

REGULATORY PERSPECTIVES

7:55 Chairperson’s Remarks

Lilia Macovei, PhD, Bioanalytical Principal Investigator, Senior Scientist, Pfizer Inc.


8:00 KEYNOTE PRESENTATION: Peptides Identified on Monocyte Derived Dendritic Cells: A Marker for Clinical Immunogenicity to FVIII

Wojciech Jankowski, Commissioner’s Fellow, CBER, FDA

The immune response to protein-therapeutics (immunogenicity) is an important safety and efficacy concern during drug development and regulation. Non-clinical assays that can be used in the early stages of clinical development and to identify at-risk individuals and subpopulations in the clinic are an important unmet need. The so-called MHC Associated Peptide Proteomic (MAPPs) assay directly identifies peptides derived from a protein of interest on a donor’s MHC-II proteins. Here we have applied this technique to address several questions related to the use of Factor VIII (FVIII) replacement therapy, in the treatment of haemophilia A. Over a dozen FVIII products are marketed, but most fall into 3 categories: (i) Purified from human plasma (PD-FVIII). (ii) Full-length FVIII manufactured using recombinant DNA technology (FL-rFVIII). (iii) A truncated, beta-domain deleted rFVIII (BDD-rFVIII). We investigated whether there are differences in the FVIII peptides found on the MHC-II proteins of the same individual when incubated with products from the 3 classes.

8:30 NEW PANEL DISCUSSION: FDA Regulatory Guidance: Essential Insight from the FDA and Industry

NEW Moderator: Jim Little, Vice President, Business Development, Thomas A. Little Consulting

Panelists:

Joao Pedras-Vasconcelos, Ph.D., Biotech Quality and Immunogenicity Reviewer, Biotechnology Products, CDER, FDA

Wojciech Jankowski, Commissioner's Fellow, CBER, FDA

NEW: Zenobia F. Taraporewala, PhD, CMC Reviewer and Team Lead, CBER, FDA 

Jim McNally, PhD, Executive Director, Head of Research and Clinical Assays, Program Lead, CRISPR Therapeutics

Shannon Chilewski, PhD, Research Scientist II, Bristol-Myers Squibb

9:20 Coffee Break in the Exhibit Hall with Poster Viewing

ASSAY DEVELOPMENT AND VALIDATION

10:10 Strategies to Implement Immunogenicity Method Updates within a Clinical Program

Chilewski_ShannonShannon Chilewski, PhD, Research Scientist II, Bristol-Myers Squibb

As part of the immunogenicity risk assessment, a Sponsor may implement different approaches to ensure data comparability after an immunogenicity method change is needed. This presentation will cover 2 case studies with distinctly different approaches.

10:30 NEW: An Evaluation of Modern and Traditional Statistical Methods for Setting ADA and NAb Immunoassay Cut Points

Jim Little, Vice President, Business Development, Thomas A. Little Consulting

This presentation will use representative ADA/NAb data to explore traditional and modern statistical methods for screening and confirmatory cut points. Logical workflow and decision trees will be presented. Published popular methods from the literature will be presented along with more modern methods of distribution fitting alternatives. Also, FDA guidance will be examined for any direction/preference to method alternatives.

IMMUNOGENICITY OF IMMUNO-ONCOLOGY DRUGS

10:50 Towards a Fit-for-Purpose Approach for Evaluating Relevant Immunogenicity of mAbs in Oncology Clinical Trials

Hassanein_MohamedMohamed Hassanein, PhD, Staff Scientist, Bioanalytical Sciences, Regeneron Pharmaceuticals

Recent data from oncology trials indicated that human mAbs (h-mAbs) elicit low treatment-emergent immunogenicity. In this presentation, we will provide a tailored approach for assessing immunogenicity of mAbs in oncology trials that factors in both the low risk profile of h-mAbs, as well as the immunity status of the target population. The goal of this “fit-for-purpose” approach is to provide immunogenicity data that is relevant to health-care providers and patients.

11:10 High-Throughput in vitro Assays for Immunogenicity Predictions and Risk Assessment

Tasker_CarleyCarley Tasker, PhD, Associate Scientist, Merck Research Laboratories

Efficient assessment of biologic-specific immunogenicity is a critical step in determining safety and efficacy. We have developed a novel in vitro assay to streamline immunogenicity assessments, allowing for high-throughput prediction of intracellular processing and presentation of potential antigenic epitopes. In a competition-based assay, soluble T cell receptors recognizing an HLA-reference peptide complex are used to detect presentation of potential immunogenic epitopes by mono-allelic, antigen-presenting cell lines.

11:40 Close of Immunogenicity Assessment & Clinical Relevance


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