The discovery and development of emerging therapies such as bispecifics, antibody-drug conjugates, biosimilars, and cell/gene therapies has raised the bar for bioassay scientists. There are numerous considerations to keep in mind during bioassay development, ranging from statistics to life cycle management. At the Fourth Annual Optimizing Bioassays for Biologics, bioassay experts will address the top challenges in bioassay design including novel technologies, increasing complex mechanisms-of-action, regulation, and the application of statistics in assay development. Case studies and best practices for handling the most common issues in biological assay design will be presented. Overall, this event will showcase ways to continue moving a biologic forward in the discovery pipeline.

Final Agenda

THURSDAY, MAY 3

OPTIMIZING BIOASSAY DESIGN
Beacon Hill Complex

12:00 pm Registration (Commonwealth Hall)

12:35 Luncheon in the Exhibit Hall with Poster Viewing (Commonwealth Hall)

1:40 Chairperson’s Remarks

Jenny Hu, Scientist, PKDM, Amgen, Inc.

1:50 Identifying and Controlling Sources of Variation in Cell-Based Potency Assays

Emily Lowe, PhD, Senior Scientist, Analytical Sciences, Kite Pharma, a Gilead Company

Cell-based potency assays are essential for engineered cell therapy products to demonstrate that drug product activity is linked to biological critical quality attributes. One of the biggest challenges in designing and executing cell-based potency assays is identifying and controlling variability. A poorly controlled and highly variable potency assay can increase invalid and re-test rates, or worse, cause a manufacturing process to appear out of control or a drug product to appear unstable. Identifying and mitigating sources of variability begins during initial assay design, as part of QbD for method development, and should continue to be a focus through life cycle management. Here, we will discuss expected and unexpected sources of variability and control strategies through presentative case studies.

2:15 USP121: Insulin Bioassay Development to Replace in vivo Rabbit Test

Dirk Usener, PhD, Head, Analytical Development 2, Bioanalytics, Sanofi

Each insulin batch has to be released for marketed use in the USA with an in vivo Bio-ID test according to USP <121>. An in vitro cell-based assay (InCellWestern) was developed and validated to substitute the in vivo assay for all batch release and stability testings, which was approved by FDA for Insulin Glargine and Glulisine. Furthermore, this in vitro cell-based assay (ICW) is planned to be implemented in USP chapter <121> which has been pre-published in USP Pharmacopoeial Forum 43(4).

2:40 Bioassays for a Bispecific Antibody Drug Conjugate (ADC)

Ashley Mullan, Scientist, Development, Analytical Sciences, MedImmune

Antibody Drug Conjugates (ADCs) are a class of biotherapeutics in which a cytotoxic agent designed to induce target cell death is conjugated to a monoclonal antibody (mAb intermediate) specific for a tumor associated antigen. This presentation will be a case study for a bi-specific ADC targeting two distinct epitopes on the target antigen. In addition to the panel of lot release assays, characterization assays developed to monitor the two different target binding epitopes will also be discussed.

3:00 Development of Cell-Based Potency Assays for Multi-Specific Antibodies

Natalia Kozhemyakina, PhD, Head, Bioassay Laboratory, Analytical Development Department, BIOCAD

Multi-specific antibody formats provide a perspective platform for the development of novel generation of more effective biotherapeutics. However, the characterization of such antibody-based multi-specifics is often made complicated by the need assays which reflect MoA of antibody fragments of which they consist. The presentation highlights automated approach, which can be applied to conduct potency screening and full analysis of candidates. It allows choose the most potent of them in the shortest period of time and as a result reduce development time.

3:20 Near-Universal Equivalence Bounds for Similarity in Bioassay

David Lansky, PhD, President, Precision Bioassay, Inc.

Testing for similarity via equivalence tests is an essential part of modern bioassay analyses. Common methods for setting equivalence bounds rely all-to-heavily on the capability of the bioassay. Sensitivity analyses show that scaled shifts in parameters measure non-similarity in ways that are assay-independent. Well-chosen equivalence bounds for scaled shifts yield assays with limited bias in potency due to non-similarity. Hence, knowledge of the required assay capability (specifically, limits on potency bias and potency trend bias) can be used to inform equivalence bounds for non-similarity. This helps is a key step towards use of an analytic target profile.

3:50 Networking Refreshment Break (Harbor & Mezzanine Level)

4:20 Development of a Reporter Gene Potency Assay for Bispecifics

Joseph Callahan, PhD, Technical Development Scientist, Genentech, Inc.

Bispecific antibodies bind two unique antigens. Here, we present a case study for a bispecific antibody reporter gene potency assay that uses two cell-types with a target cell-dependent luminescent readout. The assay was demonstrated to be MOA-reflective, robust, QC friendly, and sensitive to molecular changes in the drug. Strategies for optimization and qualification, stressed sample testing results, and efforts to enable future multi-product application will be discussed.

4:50 Adopting Multiplexing Technology for the Development of Anti-Bispecific Drug Neutralizing Antibody Bioassay

Jenny Hu, Scientist, PKDM, Amgen, Inc.

Bispecific therapeutic antibody (BsAb) recognizes two different antigen targets. BsAb usually requires separate assays to detect neutralizing antibody (NAb) response specific to each of the functional domain. Double efforts are needed for the development, optimization, and validation of two NAb assays. By utilizing multiplexing technology, we are able to develop one assay that simultaneously detects NAbs against two functional domains of BsAb.

5:20 End of Day

5:20 Registration for Dinner Short Courses (Commonwealth Hall)

Recommended Dinner Short Course(s)*

SC11: Strategic/Modular Bioassay Design and Analysis

David Lansky, PhD, President, Precision Bioassay, Inc.

*Separate registration required.

FRIDAY, MAY 4

8:00 am Morning Coffee (Harbor Level Lobby)

COLLABORATING WITH YOUR STATISTICIANS
Beacon Hill Complex

8:30 Chairperson’s Remarks

Perceval Sondag, Senior Manager, Statistics, PharmaLex

8:35 KEYNOTE PRESENTATION: Partnering with Statisticians throughout the Bioassay Lifecycle – From Clinical to Commercial

Kenneth R. Miller, PhD, Principal Scientist, Global Technical Operations, Analytical, AstraZeneca

The bioassay lifecycle consists of three stages: (1) design and development, (2) performance qualification, and (3) continued performance verification. This presentation will show how bioassay scientists and statisticians work together during each of these stages of the bioassay lifecycle. Topics will include design and evaluation of screening and robustness, design of experiments (DoE) studies, transfer of methods from development to QC, and routine monitoring of bioassays in development and QC laboratories.

9:05 The Total Error Approach Applied to Assay Validation and Transfer

Perceval Sondag, Senior Manager, Statistics, PharmaLex

Recently, the lifecycle management concept for analytical procedures was introduced. It is strongly related to the Quality by Design concept given in the ICH-Q8 guidance. This contrasts with ICH-Q2 recommendations that only focus on the validation step to evaluate the performance of an analytical procedure. ICH-Q2’s well-known check-list approach fails to provide assurance of the quality of future results with respect to the intended use of the procedure. Introduced in the early 2000s, the total error approach introduces a decision-making process based on the prediction of future results, and not on the current assay performances. Since the release of the new chapter <1210>, the USP is now onboard with this approach. This talk presents how easy it is to apply the total error approach to assay validation, and how to extend it to assay transfer. 

9:35 Simplifying Implementation of Robust Bioassays in Development to QC Lot Release

Jane Lamerdin, PhD, Director, Research & Development, Eurofins Pharma Discovery Services

Our quantitative and robust cell-based assay platform includes simple bioassays based on the native biology of the relevant receptors for potency determination & stability testing of biological drugs. These homogeneous assays employ convenient thaw-and-use cryopreserved cells to minimize assay variability and are highly scalable and suitable for automation. The talk will include bioassay qualification data for representative biosimilar and immune-oncology targets, and a bridging study to replace traditional bioassay for QC lot release.

10:05 Networking Coffee Break (Harbor & Mezzanine Level)

10:35 Review of Methods for Combining Estimates of Potency

Areti Manola, Senior Principal Biostatistician, Non-Clinical Statistics, Manufacturing, Toxicology and Applied Statistical Sciences, Janssen, Pharmaceutical Companies of Johnson and Johnson

Biopharmaceutical products require as a condition for marketing a statement on the potency of the lot and its adherence to specifications, for example 80%LC <= potency estimate <= 125%LC. There may also be a specification on the confidence interval of the potency estimate. The potency estimate may arise from a bioassay procedure that yields multiple estimates of potency. Combining these individual estimates into a single pooled estimate is an important statistical practice. Combining potency estimates is also necessary during method validation and calibration of standards. Finney (1978) and Bliss (1952) have given extensive discussions on combining potency estimates. The USP and EP have also included a section on combining estimates. In this talk, we will review these methods and emphasize that none of these references have dealt with the question of correlated estimates. We will propose an approach within the framework of a hierarchical model and compare with standard and regulatory approaches.

11:05 Everything You Always Wanted to Know about Relative Potency Bioassays… but Were Afraid to Ask

Gaël Debauve, PhD, Associate Director, Bioassay Development, Analytical Sciences for Biologics, UCB

Biological activity is measured through appropriate relative potency methods. Because of the inherent variability of biological test systems, an absolute measure of potency is more variable than a measure of activity relative to a standard. This has led to the adoption of new statistical tools addressing bioassay specificities. Though case studies, we will illustrate how USP1033 validation requirements were tackled and even exceeded using, for example, total error and variance decomposition analysis. Finally, we will provide some insights to the question: “Is my bioassay stability indicating?”

11:35 PANEL DISCUSSION: Challenges in Applying Statistics to Bioassay Design

Moderator: Perceval Sondag, Senior Manager, Statistics, PharmaLex

Panelists: Kenneth R. Miller, PhD, Principal Scientist, Global Technical Operations, Analytical, AstraZeneca

Gaël Debauve, PhD, Associate Director, Bioassay Development, Analytical Sciences for Biologics, UCB

Areti Manola, Senior Principal Biostatistician, Non-Clinical Statistics, Manufacturing, Toxicology and Applied Statistical Sciences, Janssen, Pharmaceutical Companies of Johnson and Johnson

Steven Walfish, Principal Scientific Liaison, USP

• Applying design of experiments (DoE) approaches
• Optimizing bioassay design using statistics
• Fostering collaborations between statisticians and bioassay scientists
• Application of the USP Bioassay Chapters

12:35 pm Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on your Own

1:05 Networking Refreshment Break (Harbor & Mezzanine Level)

BIOASSAY INNOVATIONS
Beacon Hill Complex

1:35 Chairperson’s Remarks

Steven Walfish, Principal Scientific Liaison, USP

1:40 The USP Bioassay Chapters: The Honeymoon’s Over?

Steven Walfish, Principal Scientific Liaison, USP

USP Chapters <111>, <1030>, <1032>, <1033> and <1034> were last updated nearly ten years ago. The USP Statistics Expert Committee is performing a periodic review of the USP Chapters to determine if there is any new advancement that would make the bioassay chapters more valuable to their stakeholders. This presentation focuses on the USP suite of bioassay chapters. A discussion of the current state, and future plans will be shared with the participants. An open dialogue with participants is expected to gain valuable feedback on areas where the chapters can be improved to be more user-friendly. Do not miss your chance to be part of the change.

2:10 A New Class of International Standards to Support Biotherapeutic Monoclonal Antibody Products Quality and Consistency over Time

Sandra Prior, PhD, Senior Scientist, Biotherapeutics, National Institute for Biological Standards and Control (NIBSC)

The approval of biosimilar biotherapeutic monoclonal antibodies (mAbs) is controlled by robust regulatory processes. However, in an increasingly complex multi-product market place there is a need for public biological standards to support current controls. With this aim, we have developed the first World Health Organization (WHO) mAb potency international standard (IS) to support the performance and calibration of bioassays and local standards. These ISs are stable lyophilized preparations that define international units (IU) of bioactivity allowing data harmonization amongst stakeholders and promote product consistency overtime.

2:40 Thaw-and-Use Target Cells Pre-Labeled with Calcein AM for Antibody Dependent Cell-Mediated Cytotoxicity Assays

Shan Chung, PhD, Principal Scientist and Group Leader, Bioanalytical Sciences, Genenetech, Inc.

This presentation will describe the development and implementation of thaw-and-use pre-labeled target cells for in vitro antibody-dependent cell-mediated cytotoxicity (ADCC) assays. Cells were pre-labeled with the fluorescent dye calcein AM, cryopreserved in single-use aliquots, and used directly in assays after thawing. Compared to freshly labeled cells, these cells showed comparable viability, label retention, and reactivity to ADCC mediated by various effector cells, and provided favorable precision and accuracy, as well as improved consistency and robustness to ADCC assays.

3:10 The Ins and Outs of Automating Potency Assays

Adrienne Wildt, PhD, Associate Director, Bioanalytical Sciences, Immunogen

Routine assays with repetitive steps are major components of bioanalytical support. Automating these tasks increases throughput, reduces performance variations and decreases repetitive strain. Strategies for automating bioassays using various robotic liquid handling solutions used during CMC product development will be presented. Case studies on automating biochemical as well as cell-based assay that support drug substance release and characterization will be discussed.

3:40 End of Conference