Biological activity is a critical authenticity and quality attribute of biologics. It is extremely important to be sure that we create methods that reflect molecule mode of action, especially in case of multiple MoA, and at the same time is sufficiently precise and accurate. It is well known how potency assay can be variable, laborious, and time consuming with its long multi-stage protocols. The presentation provides case studies of development strategies for innovative molecules and highlights best practices for handling the most common issues in biological assay development.

Standards are essential to the development and control of biological products. Considering their importance, there is no consensus on the source of a standard, the basis and means of standard qualification, and stability evaluation. This talk will discuss principles and practices related to standards used to report potency of biological products and propose strategies. Those proposals will borrow from practices related to quality by design, highlighting fitness-for-use of a standard.

This discussion will cover standards for regenerative medicine products. Discussion will focus on standards for bioassays, published standards and how to implement these standards be addressed. An overview of current standards under development and how to get involved, as well as a forum for questions will be provided.

The Biologics Price Competition and Innovation Act was passed as part of the Affordable Care Act and under that law, insulins will be regulated as biologics in March 2020. Under the regulations for Biologics License Application review, it is expected that the potency of the insulins and of their associated biosimilars will be assessed quantitatively in a cell-based assay or bioassay that, ideally, represents the product mechanism of action. Currently, the USP is recommending an in vivo rabbit bioidentity test for the assessment of the biological activity of insulins. However, reduction in animal experiments is a worldwide goal for many and it is likely that in vitro bioassays will be submitted for assessing the potency of insulins products. This research talk will highlight some of the aspects of metabolic functional assays to think about during development and product characterization for insulin products and for antibodies that target metabolic pathways.

A rabbit blood sugar bioidentity assay is required by the FDA to evaluate biological activity for insulin analogs per USP<121> guideline. Not only are a large number of live animals used, but the method is also highly variable and expensive. A reporter cell-based assay was developed by utilizing insulin’s role in regulating hepatic gluconeogenesis pathway. This functional assay was qualified to demonstrate its performance in linearity, accuracy and precision, and was further evaluated on its robustness using design-of-experiment (DoE) approach. Lastly, it was compared with an established insulin receptor phosphorylation assay to demonstrate its superior performance.

Relative potency is a measure obtained from the comparison of a test sample to a standard based on the capacity to produce the expected biological activity. USP <1034> presents several different models and suitability criteria to determine the reliability of the estimate. This talk will cover traditional linear and non-linear bioassay models with an emphasis on model suitability including parallelism.

At the heart of bioassays for biologics are critical reagents used to directly or indirectly measure biologic markers or signals. A comprehensive analytical toolbox of biochemical, functional and biophysical methods (UPLC-SEC, IEX, RP, HIC, CE, icIEF, Octet, and HRMS) has been developed to evaluate the quality of critical reagents. Several case studies, including reagents, troubleshooting and comparison of small scale hybridoma and large-scale recombinant protein, lifecycle management of biologics, developing reference material, emerging standards and regulatory frameworks, will be presented.