Bioassays 2020: Scientific Approaches and Regulatory Strategies

Analytics play a critical role in assuring product quality through release testing, process control, deep characterization, defining critical quality attributes, comparability and analytical similarity. A framework that would facilitate the use of this information and allow for effective knowledge management will enhance quality control strategies and drug development. This concept is explored. Bioassays are a key part of analytics for quality; Such assays and biological models in general can also facilitate effective clinical development of drugs. Communication and structures that leverage biology across disciplines may enhance drug development.

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The technical aspects of bioassay strategies are essential as ever while the biotech industry launches new biologics including modified proteins, DNA, RNA and cell-based therapeutics. The focus of this session will reinforce the importance of those bioassay technical features that contribute to the successful performance of a bioassay from assay development through to the post-marketing stage. Some of the common threads intersecting bioassay applications and strategies that will be discussed during this session include bioassay design considerations, critical reagents (e.g. reference standards), data analysis approaches, and control measures used throughout the product lifecycle.

DoE is a crucial part during the development of potency assays at the Potency Assay Skill Center of Boehringer Ingelheim. The DoE approach is accelerating assay development as it allows the combination of several development experiments in one set. Therefore the total number and extend of assay runs needed is significantly reduced. Furthermore, it increases assay quality as it e.g. helps to identify 2-factor interactions which are only hardly detectable with the OFAT (one factor at the time) approach. Automation of DoEs is done to reduce variability of potency assays which improves the output of the DoEs. The process of DoE automation was split into two parts: Semi-automation of pipetting steps have been introduced in DoE experiments by using automated electronic multichannel pipettes. To further minimize handling time and to improve the assay throughput, fully automation of DoE by using an automation workstation is being established. Opportunities and challenges of the semi and fully automated DoE approaches will be shown.

The technical aspects of bioassay strategies are essential as ever while the biotech industry launches new biologics including modified proteins, DNA, RNA and cell-based therapeutics. The focus of this session will reinforce the importance of those bioassay technical features that contribute to the successful performance of a bioassay from assay development through to the post-marketing stage. Some of the common threads intersecting bioassay applications and strategies that will be discussed during this session include bioassay design considerations, critical reagents (e.g. reference standards), data analysis approaches, and control measures used throughout the product lifecycle.

Experience shows that the handling of compendial and especially non-compendial and in-house reference standards and materials are often a challenge for OMCLs (Official Medicines Control Labs). The qualification of biological reference materials by the manufacturer and the OMCL has a direct influence on the final results of analytical tests and is thus of extremely high importance. The replacement of the reference vaccine’ case study highlights the difficulties encountered by the manufacturers and National Control Labs dealing with biological materials and methods(e.g. :In vivo, ELISA), and how to overcome them.

Potency determination via bioassay is a relative measure that requires an evaluation of parallelism between the dose-response relationships of a reference standard and sample material. Typical approaches for assessing parallelism include difference (p-value) and equivalence tests. These traditional methods rely on a statistical assessment of model parameters as opposed to direct evaluation of the similarity of the dose response curves. A simple and intuitive curve similarity approach is proposed that tests the hypothesis that the sample material is a dilution or concentration of the reference standard. The test is achieved by quantifying the total area between the two curves and provides a single composite measure of parallelism that ranges from 0 to 1. Both a frequentist and a Bayesian approach to the test are provided. A simulation study shows that the curve similarity approach overcomes the drawbacks of the typical methods and is effective at detecting parallelism and non-parallelism for bioassays.

Catalent Biologics provides advanced technologies and solutions for protein and gene therapy development, manufacturing and clinical supply. With technology platforms including GPEx® cell line engineering, SMARTag® bioconjugation and AAV viral vector expression combined with comprehensive analytical services and fill/finish supply, Catalent Biologics is your premier partner for clinical and commercial success.

With increasing numbers of cell and gene therapy products in development and entering commercialization, more and more sponsors are facing a unique set of challenges of developing bioassays for cell and gene therapy products. Developing appropriate bioassays for cell and gene therapy products tends to be significantly more difficult than for conventional biologics due to a number of different factors, including- the difficulty of adequately modelling the clinical mode(s) of action in a simple cellular system; the challenge of defining, assessing suitability and generating sufficient quantity of, reference substances; non-linear or non-sigmoidal dose response relationships in some bioassays; qualitative or only semi-quantitative responses in others; a need for rapid analytical turnaround times in some cases; an evolving understanding of clinical mode(s) of action in some cases, which may require a close interplay between analytical and clinical development; and finally, the challenge of selecting the best bioassay(s) for release and stability testing vs. characterization/comparability testing. This session will focus on case studies and regulatory feedback that we hope will be beneficial to all sponsors faced with the daunting task of developing a comprehensive control strategy for a cell or gene therapy product.

The development of a relative potency assay to support biologic drug products requires significant budget, resources, and time to achieve the desired quality for release testing. Potency assays ensure safe and efficacious products are delivered to patients, and cell-based bioassays require 1) a relevant and permissive cell line, 2) ability to demonstrate the drug’s mechanism of action (MoA), and 3) quantitative end point. Cell line engineering can achieve these requirements and simplify the MoA detection in a reporter gene system. Here we describe a case study where an engineered reporter gene cell line was used to develop and qualify a relative potency assay within Analytical Development. A cross-over study was successfully performed to transition assay platforms during an ongoing Phase I GMP stability study.

Ultragenyx Gene Therapy is focused on advancing AAV-based gene therapies for rare, but serious, genetic diseases. Potency assessment for such advanced therapeutic product is of critical importance, but can also be very challenging. Because a reliable potency assay is required as early as Phase I/II of product development, we have developed a relative potency assay by measuring the expression of transgene specific mRNA after the transductions of appropriate target cells. Assay was miniaturized and multiplexing was implemented to improve assay throughput and performance. This potency assay shows a high level of robustness and throughput, and requires only minor adjustment while modifying it for novel products. This presentation describes how this platform potency assay was developed and successfully adapted for two different gene therapy products.

We share your passion for science. Promega is committed to excellence in cell-based assays and protein characterization tools for biologics - from lead generation to lot release. Our bioassays for ADCC, immune checkpoints and biosimilars, plus our mass spec proteases and reagents, are used widely throughout the industry. Let’s collaborate.