Mastering the biosafety of biologics from early phase to successful IND and BLA filings

Noah Horn, Manager, Viral Clearance Services; Lara Strittmatter, Manager, BioPharma Cell and Gene Therapy, EBPT Lancaster

Because biological products rely on complex nutrient and matrix needs of the living systems required to produce them, extraneous contamination events, either through accidental introduction (adventitious) or endogenous to the cell line, pose substantial risk to the end-point purity of a product. To confirm the absence of adventitious agents and demonstrate control of the manufacturing process, samples of unclarified bioreactor material, termed Unprocessed Bulk (UPB), are required to undergo confirmatory and quantitative viral and microbial testing. EBPT’s Lancaster site offers UPB testing services which include detection via in vitro Adventitious Agent (IVAA) testing, quantitative GMP Transmission Electron Microscopy (TEM) for retroviral viral-like particles (RVLPs) enumeration, mycoplasma testing, bioburden analysis, MMV (minute virus of mice) targeted-PCR, as well as confirmation of safe product generation with viral clearance validation.

Viral safety is a major regulatory expectation for biological products and can lead to review and approval delays by agencies, such as the FDA, if not adequately addressed. Biological products are cultivated in environments that may pose diverse risks. IVAA testing remains an important aspect in determining the safety profile of any biologic product produced in bioreactors. IVAA is tasked with confirmation of whether contaminating virus species are present in every manufactured lot, but the test does not determine the strain or quantity of virus present. This work is performed by applying the product to a variety of cells from different biological origins, including human and cells of the same species as those used to produce the biologic. Once inoculated, the cells are incubated under conditions and time periods sufficient to allow growth and detection of any contaminating virus. In addition to the traditional IVAA approach of live cell assays, EBPT is expanding its capabilities with Next Generation Sequencing (NGS) to detect viral contaminants in product matrices that may not be suitable for in vitro analysis or that require a shorter turnaround time before patient administration.

For cell lines containing endogenous retroviruses, regulatory standards require the removal of RLVPs from the final product. Thus, accurate quantification of these endogenous RLVPs is essential for determining the Log Reduction Value (LRV) of the harvest material. In 2023, EBPT Lancaster implemented an in-house GMP method for the quantification of RVLPs using thin-sectioned TEM. This allows handling of all UPB testing by a single, coordinated biosafety team at one site. In addition, during the validation of this method, we increased efficiency through an improved embedding method, automated imaging, and a streamlined results screening approach. As a result, we have significantly reduced turnaround times making our process more competitive and can offer all UPB testing results within 35 days. The LRV of three lots obtained via this method is used to inform the requirements of the viral clearance demonstration.

Viral clearance testing evaluates the capabilities of the production pipeline purification processes to determine if viral contaminants could be successfully destroyed and removed prior to fill-finish execution. To meet this vital safety requirement, EBPT’s Viral Clearance Team produces and validates virus lots in-house ensuring that each client’s purification process is robustly challenged. To empower the evaluation of each product, every project is customized to meet the target LRV. EBPT’s viral clearance facility includes fully equipped, specialized suites for process execution and evaluation while providing staff with expertise to perform these critical tests to include in our client’s Investigative New Drug (IND) and Biological Licensing Application (BLA) filings. The Viral Clearance Department continues to expand its capabilities by adding new testing options.

Facilitating the integration of new products and supporting technologies to further improve global health and life expectancy is what drives Eurofins BPT. With our partners, we strive to provide life-saving products to the people that need them. The Biosafety teams at EBPT’s Lancaster, PA, facility have the resources and technical expertise to support the growing diversity of biologic products. Our capabilities support clients from early phase development through IND and BLA filings and beyond.

EBPT’s Biosafety Team utilizes a breadth of technical experience and regulatory familiarity to perform safety testing critical to the development of new biologically based treatments alongside the continuing technological developments in the biopharmaceutical industry. The Biosafety teams’ capabilities encapsulate the full scope of testing needs from cell bank preparation and cell line characterization to its safety validation with TEM, IVAA, and viral clearance. All these services are centered on integrating customization and flexibility to best serve our clients, and ultimately the patients that depend on the safety of the products they need.