Optimizing Bioassays for Biologics

Demonstrating biological activity in a drug is critical, yet is becoming increasingly difficult as the industry moves towards use of molecules with multiple mechanisms such as bispecifics and antibody drug conjugates (ADCs). Along with this complexity there are numerous considerations: from statistics to life cycle management. At the Second Annual Optimizing Bioassays for Biologics, industry experts will discuss the most pressing bottlenecks in bioassay management beginning with novel technologies and techniques and moving through to assay bridging and transfer. High throughput, high density approaches will be emphasized at this year’s event and solutions for optimizing sensitivity, specificity, and selectivity will be discussed. The use of design of experiment (DoE) will be integrated throughout to provide delegates with best practices for moving a biologic forward in the discovery pipeline. 

Recommended Dinner Short Course*

SC9: Overcoming the Challenges of Immunogenicity Assessment

*Separate registration required.




12:35 pm Luncheon in the Exhibit Hall with Poster Viewing

1:40 Chairperson’s Remarks

Tilman Schlothauer, Ph.D., Principal Scientist, Large Molecule Research, Roche Innovation Center

1:50 Potency Assay Development and Associated Extended Characterization Strategy of Therapeutic mAb

Tilman_SchlothauerTilman Schlothauer, Ph.D., Principal Scientist, Large Molecule Research, Roche Innovation Center

During the past years we have collected several examples of profound structure function correlations for IgG-based therapeutic modality development. These insights have been used as the basis for the following Potency Assay strategy. Applying a set of orthogonal cell-based and cell free functional assays always provides a deep understanding of the molecule and related critical quality attributes.

2:20 Reporter-Gene Assay for the Measurement of Functional Activity and Neutralizing Antibody Response to Infliximab: 3 Years of Clinical Laboratory Application

Julio Delgado, M.D., M.S. Tenured Associate Professor, Pathology, University of Utah School of Medicine; Section Chief, Immunology Division, ARUP Laboratories

TNF-α antagonists such as infliximab are effective for the treatment of inflammatory bowel disease and other inflammatory and autoimmune diseases. Development of an immune response and subsequent antibodies against these protein-based drugs is a major impediment that contributes to therapeutic failure, or adverse effects such as hypersensitivity reactions. Rational and cost-effective evaluation of therapeutic failure includes measurement of serum drug levels, and detection of drug-specific antibodies. Reporter-Gene Assay (RGA) methodology is a robust platform for measuring biologically active serum levels of TNF-alpha antagonists, and for the detection of neutralizing antibody responses. The RGA assay allows for high-throughput testing, which makes it well suited for the clinical laboratory. At this point, this assay is primarily indicated for the management of patients developing treatment failure. Prospective randomized studies are necessary to establish appropriate medical decision intervals for therapeutic drug monitoring using this assay.

2:50 Development of Luminex as a Platform for the Detection of Anti-Drug Antibody IgE Isotypes in Human Serum

LiNa_LooLiNa Loo, Ph.D., Associate Principal Scientist, Bioanalytical Development, Merck

Since biotherapeutic drugs such as monoclonal antibodies (mAbs) have the potential to induce immunogenicity, it is critical to perform an immunogenicity assessment to ensure drug efficacy and patient safety. Here, Luminex and Mesoscale were evaluated as platforms for detection of anti-drug antibody IgE isotype in human serum. By using a mouse-human chimeric drug-specific monoclonal IgE antibody as the positive control, the assay characteristics were compared for the two platforms.

3:20 Predicting Adverse Immune Responses to Biologics

Anne_DickinsonAnne Dickinson, Ph.D., Professor, Marrow Transplant Biology, Newcastle University

There are currently no reliable human in vitro assays which test for immunogenicity, sensitivity, efficacy and allergic reactions of biologics that are equivalent to or superior to in vivo animal testing. Alcyomics has developed a novel test Skimune™, a non-artificial (non-3D) human in vitro skin test which can predict allergic or adverse immune reactions to test compounds. The test gives a predictive readout of graded histopathological skin damage and has been shown to correlate with inflammatory cytokine release and T cell proliferation responses. We have further developed this assay to investigate monoclonal antibodies (Skimune™Mab) and antibody drug conjugates (Skimune™Mab-antibody drug conjugate.

3:50 Refreshment Break


4:20 Potency Assay Selection for Antibody-Drug Conjugates: Challenges and Considerations

Shelly_ElvingtonShelley Elvington, Ph.D., Technical Development Scientist, Genentech

Antibody-drug conjugates combine the target specificity of a monoclonal antibody with the activity of a potent therapeutic compound. This unique format leads to special considerations when developing potency assays. Here, I will present our strategy for ADC assay development, including selection of appropriate formats and cell lines that take into account the unique properties of the ADC (including impact of drug-antibody ratio).

4:50 Bioassay Strategy and Challenges for a kλ Body - A Next Generation Bispecific

Anaelle_DosSantosAnaëlle Dos Santos, Head, Bioanalytics Unit, NovImmune SA

kλ bodies are a novel bispecific format with proprieties indistinguishable from an IgG. The strategy will be presented for development of a potency assay for a kλ body which has a carefully selected affinity for CD47 in order to selectively block this receptor in B cell malignancies. A comprehensive suite of highly sensitive binding and reporter gene assays have been developed and qualified to assess the potency of the product in batch release, stability and characterization studies. The advantages and limitations of each of these assays will be discussed.

5:20 End of Day

5:15 Registration for Dinner Short Courses

Recommended Dinner Short Course*

SC12: Strategic Bioassay Design and Analysis

*Separate registration required.


8:00 am Registration and Morning Coffee

8:30 Chairperson’s Remarks

Shelley Elvington, Ph.D., Technical Development Scientist, Genentech

8:35 PANEL DISCUSSION: Bioassays for the Changing Biologics Landscape

Moderator: Shelley Elvington, Ph.D., Technical Development Scientist, Genentech

Panelists: Tilman Schlothauer, Ph.D., Principal Scientist, Large Molecule Research, Roche Innovation Center

  • Immuno-oncology bioassays
  • Antibody-drug conjugates
  • Bispecific antibodies


9:35 Successful Assay Validation – The Role of Sample Size Calculations to Ensure Regulatory Acceptance

Walter_HoyerWalter Hoyer, Ph.D., Principal Statistician,CMC Statistics, GSK Vaccines GmbH

In recent years regulatory authorities have strongly increased demand for (bio-)analytical method validations. Two recurring themes are “variance component analysis” to assess intermediate precision and more generally the preference for equivalence tests over the classical nullhypothesis testing approach, as outlined in USP. The presentation combines both aspects and presents a pragmatic way of performing power and sample-size calculations for the total variance of general multi-factor mixed models. The approach allows to enter an assay validation with confidence to pass and yet satisfy the stringent requirements of regulatory authorities.

10:05 Coffee Break


10:35 Mitigating Variability in a Non-Homogenous Assay Format-A Case Study

Linda Collins, Senior Associate Scientist, Functional Biocharacterization, Amgen

Selection of the assay format, inclusive of the plate type, is a critical step during bioassay development. This is particularly true when the assay is a non-homogeneous format that includes washing steps. This talk will present a case study of a cell-based potency assay that was developed with a specialized plate type. A change in the manufacturing process led to a change in the plate quality with a corresponding decline in assay performance. The efforts undertaken to mitigate the decline in method performance will be discussed. 

11:05 Optimization of a Potency Assay Using DoE in Support of Monoclonal Antibody Product Development

Arden Bond, Staff Scientist, Analytical Development, Genzyme

This presentation describes the use of Design of Experiment (DoE) to evaluate the method performance of a cell-based potency assay. DoE is a technique for designing and analyzing experiments, using a minimum number of assays by systematically varying several parameters simultaneously. After initial development of a cell-based potency assay, DoE was used to confirm method accuracy, range, sensitivity and robustness. Once appropriate method performance was demonstrated, the cell-based potency assay was used to support product development. The potency assay method design and optimization will be discussed.

11:35 pm Optimization and Robustness Study of a Potency Assay Using DoE

Jan_AmstrupJan Amstrup, Ph.D., Principal Scientist, CMC Bioassay, Novo Nordisk A/S

Optimization of a potency assay by use of DoE in the optimization process will be shown. Nine assay factors and four factor interactions based on time consumption and complexity of buffers were evaluated. Results obtained from the DoE provided improved assay conditions and settings that were successfully evaluated in a proof-of-concept assay. Furthermore, how to implement the optimizations will be discussed with regard to validation status of the assay.

12:05 pm Enjoy Lunch on Your Own

1:05 Refreshment Break

1:35 Chairperson’s Remarks

Gaël Debauve, Ph.D., Analytical Sciences for Biologicals - Bioassay Development Laboratory, UCB Pharma S.A., Braine-L’Alleud

1:40 PANEL DISCUSSION: DoE for Biological Assays

Moderator: Gaël Debauve, Ph.D., Analytical Sciences for Biologicals - Bioassay Development Laboratory, UCB Pharma S.A., Braine-L’Alleud

Panelists: Linda Collins, Senior Associate Scientist, Functional Biocharacterization, Amgen

Arden Bond, Staff Scientist, Analytical Development, Genzyme

Jan Amstrup, Ph.D., Principal Scientist, CMC Bioassay, Novo Nordisk A/S

  • Why? Business and regulatory expectations
  • How? Pro/cons, limitations and challenges
  • When/What? DoE integration into method lifecycle: When is it implemented?


2:40 Bioassay Lifecycle: From Development to Continuous Verification

Gael_DebauveGaël Debauve, Ph.D. Analytical Sciences for Biologicals - Bioassay Development Laboratory, UCB Pharma S.A., Braine-L’Alleud

Biological activity is a critical quality attribute for biopharmaceutical products and cell-based bioassays are generally used to accurately determine this activity. Classical proliferation assay presents a lot of drawbacks that could be overcome by the gene reporter technology (e.g. in terms of method simplicity, reduced variability and improved lead time). This presentation will go through a case study illustrating the gene reporter journey: from the decision to switch to a gene reporter assay to the tools implemented to monitor the method performance.

3:10 Determination of the Limit of Detection and the Limit of Quantitation during Assay Development in the Biopharmaceutical Industry

Eloi_KpameganEloi P. Kpamegan, Ph.D., MSF, Executive Director, Clinical & Nonclinical Biostatistics, Novavax, Inc.

The usefulness and optimal throughput of an assay may depend on the appropriate determination of the LOD and the LOQ. The experiment design and statistical analysis method used for the determination of LOD and LOQ are dependent on the assay type (e.g., ELISA, Functional or PCR). This presentation describes the design, testing and statistical procedures required to determine the LOD and LOQ during assay development. The procedures to be used to confirm the LOD and the LOQ during assay validation are discussed.

3:40 End of Optimizing Bioassays for Biologics