Viral Vaccine Process Development

Guiding Principles In Process Development For A Viral Vaccine

At Batavia Biosciences, our approach to viral vaccine process development is underscored by three fundamental guiding tenets. Herein, we illustrate these principles:

Optimized Process Development from the Outset

Our holistic strategy for the inception of a production protocol aims to ensure a seamless transition into phase I and II clinical trials. This strategy is deeply rooted in a comprehensive grasp of the prospective commercial manufacturing process, from the raw materials used to the equipment and process structure earmarked for clinical production.

The objective is to ensure a smooth transition to commercial-scale manufacturing. Any oversight in this step might warrant considerable process alterations down the line, potentially causing regulatory bodies to demand new clinical trials for validating safety and efficacy. Drawing from our wealth of experience in constructing detailed and pragmatic product development plans within the viral vaccine industry, we guide our clients in choosing the right pathway for process development. This contributes to a more efficient product-to-patient delivery and increases shareholder value, a vital factor given the ever-evolving market dynamics, patient demographics, and dosing regimens in the viral vaccine landscape.

Cost-effective Viral Vaccine Production

Considering the substantial influence of manufacturing costs on the product’s overall price, establishing a cost-efficient production process is crucial. This includes focusing on key elements like the pre-culture process to optimize production bioreactor utilization, ideal bioreactor process yield (USP), and purification recovery (DSP). These aspects greatly impact the Cost of Goods (CoG) and necessitate a stringent evaluation to identify optimal output parameters and acceptable variances.

These parameters then serve as performance metrics during process development (design space). We offer in silico cost modeling services and have a proven track record of aligning process development with critical economic parameters. Our comprehensive suite of technologies and capabilities plays a vital role in reducing CoG and development timelines.

 

Robust Documentation Practices

Since the inception of Batavia Biosciences, we’ve understood the pivotal role of the Quality Assurance (QA) department in R&D project dialogues. This realization has significantly contributed to our successes in IND and IMPD support. We underscore the importance of early QA involvement through three initiatives designed to streamline the clinical trial process:

Planning Phase: Our customers universally seek strategies to expedite the clinical trial process or to defer product development investments. In response, our QA department collaborates with technical staff and customers in constructing comprehensive product development plans. These plans justify, outline, and establish risk-based decisions on the most effective route to the clinic.

Quality Management System: Within the R&D environment, all selected materials and equipment are vetted for GMP manufacturing suitability, ensuring a seamless transition of the developed process to a GMP facility without any redundant process elements. Additionally, the Quality Management system adopted in R&D mirrors that used in GMP.

Internal Tech Transfer Elimination: Our laboratory personnel involved in R&D also possess the requisite training and qualifications to operate within a GMP environment. Consequently, the team responsible for developing production and purification processes in R&D is also fully equipped to transition the project into GMP. This approach eliminates time-consuming internal technology transfer processes that would otherwise be mandatory.

By adhering to these guiding principles, Batavia Biosciences strives to consistently deliver on viral vaccine process development, effectively serving the broader scientific community.

Upstream Process Development For Vaccine Production

Our operations span various production scales and employ a wide array of cell culture apparatus for both adherent and suspension growth. Our scales extend from high-efficiency 20 mL bioreactors to larger 200 L systems. Our assortment of equipment includes shake flasks, cell factories, WAVE Bioreactors™, and an array of stirred tank bioreactors from multiple manufacturers.

In addition, we utilize iCELLis® technology for process development (iCELLis® nano) and GMP production (up to iCELLis® 500). Our latest addition to the manufacturing platforms is the innovative scale-X™ high cell density bioreactor, which offers similar output to the iCELLis® but features a significantly reduced design footprint, substantially decreasing production space requirements.

Viral Vaccine Production Titers
We conduct state-of-the-art manufacturing of a diverse range of viruses, attaining standards in line with published literature, using adherent cell culture, suspension culture, and microcarrier-based processes. For instance, utilizing the Vero cell line for H1N1 influenza virus production, we have achieved high virus titers (1 x 1010 TCID50/mL) in 5 L bioreactors, consistent with the findings documented in the literature (Chen et al., 2011). Furthermore, in the production of the measles virus using bioreactors, we typically reach a peak titer of approximately 5 x 107 TCID50/mL, as reported in the literature (Weiss et al., 2012).

Developing Upstream Process Protocols
We employ our standard manufacturing protocols and our SCOUT® technology for the development of product-specific protocols. This includes testing a range of commercially available production media and standard feed strategies in mini-bioreactors to optimize cell growth, shorten population doubling times, and increase cell densities.

Downstream Process Development For Viral Vaccines

At Batavia Biosciences, we harness appropriately scaled downstream equipment that aligns with the various upstream scales. These tools facilitate clarification, concentration, chromatography, and filtration. To purify viruses, we offer clarification through depth filtration utilizing a range of filters, including those that synergistically facilitate particle purification and impurity removal.

Often, viral products necessitate the removal of nucleic acids, which can be accomplished through techniques such as enzymatic digestion or fractional precipitation. Preceding the purification phase, processes like concentration and buffer exchange can be conducted using tangential flow filtration (TFF), implemented via hollow fibers or flat screens. The choice between these two is dictated by the target virus’s physical properties and the final product’s specifications.

Purification is typically realized through several chromatography steps, the nature of which depends on the virus’s properties. Purification processes commonly exploit differences in ion exchange or hydrophobic interaction. Size-based group separation or TFF can be employed for polishing and formulation, utilizing hollow fibers or flat screens. A sterile filtration can be executed for viral products of a specific size. We leverage our vast experience in validated aseptic manufacturing processes for larger viruses where sterile filtration is not viable. For product analysis, we provide an array of product-specific assays and assay development services, including TCID50, PFA, FFA, ELISA, (q)PCR, and HPLC-based methods.

Downstream Processing Equipment for Viral Vaccines

Our proficiency extends to developing virus purification processes from the ground up, expeditious transfer of existing processes, and enhancement of existing processes to augment purity or recovery. Initiating a process from scratch often involves small-scale clarification development, employing scalable filters.

Minor ultrafiltration/diafiltration (UF/DF) development for concentration and buffer exchange can be conducted if necessary. This is followed by chromatography development, beginning with selecting resins for binding and elution profiles.

This evolution extends from 96-well plate screening to 1 mL spin traps and spin filters, progressing to intermediate and large-scale purification columns/filters/monoliths, facilitated by ÄKTA™ explorer, ÄKTA™ pure, and ÄKTA™ pilot. A typical viral vaccine purification process usually necessitates a two- to six-step process, yielding 35-80% product recovery while maintaining compliance with purity regulations.

A conventional viral vaccine purification process encompasses clarification and concentration steps—occasionally preceded by cell lysis and DNA removal—followed by chromatography, employing charge, hydrophobic interaction, mixed mode, size exclusion, or affinity-based techniques.

Ultimately, the vaccine is formulated by re-buffering and adding components, succeeded by a final membrane filtration step utilizing a bioburden reduction or sterilizing grade filter. We predominantly use centrifugation-based methods to deliver research batches for preclinical studies to ensure highly pure virus preparations.

Analytical Development For Viral Vaccines

The evolution of virus manufacturing processes is deeply interlinked with the accessibility of precise and reproducible product characterization and release assays. These critical release tests for viral products typically encompass measurements of quantity, potency, genetic stability, identity, residuals (e.g., host cell DNA, host cell protein, and benzonase), and safety.

The product characterization stage is imperative to discern the parameters that necessitate close monitoring during process development and those essential for product release. Most techniques and assays utilized for this characterization process are performed in-house at Batavia Biosciences.

Specific assays are developed to support process development using representative material, which is subsequently validated for final production release. Our analytical staff, with their profound expertise, oversee every facet, from the design, implementation, and development to the validation of assays, in strict adherence to ICH guidelines. Some compendial and most biosafety assays are outsourced to our qualified partners, with Batavia Biosciences providing comprehensive support across all areas, including partner selection, quotations, temperature-controlled shipments, review, and reporting in certificates of analysis.

Controlling Materials for Viral Vaccine Production

Rigorous control over the materials used to manufacture viral vaccines is indispensable. The usage of animal-derived components is avoided to the greatest extent possible, and when unavoidable, traceability and testing measures are implemented to mitigate any risks associated with introducing and transferring extraneous agents in the process. Comprehensive testing, such as PCR for in vitro and in vivo adventitious agents, ensures the absence of adventitious viruses in master cell banks (MCB) and master virus seeds (MVS), per ICH Q5A and ICH Q5D guidelines.

Genetic Stability Testing

Monitoring genetic stability stands as another main parameter in virus production. This procedure propagates the virus through several passages until or beyond the envisioned commercial manufacturing stage, as guided by general human vaccine considerations (http://www.usp.org/).

The analysis of genetic content is crucial to ensure the strain’s stability throughout the propagation process. Extended propagation of the starting material provides the sensitivity to detect potential recombinants or mutants that may possess a growth advantage over the target strain. PCR detection and sequence analysis offer the necessary specificity and sensitivity to identify emerging transgene region variants. Moreover, next-generation sequencing technologies can be harnessed to characterize virus variants potentially present at low frequencies in vaccine preparations or virus seeds.

Analytical Development Expertise

Batavia Biosciences boasts extensive experience in the development and qualification of assays, encompassing methodologies such as (q)PCR, cell-based assays (FFA, PFA, and TCID50), HPLC, SDS-PAGE, western blot, and ELISA. Consequently, we can deploy a comprehensive range of assays vital for successfully monitoring the development and release of viral vaccines.

In Silico Cost Modeling For Viral Vaccines: Optimizing Manufacturing Processes

At Batavia Biosciences, we frequently employ in silico cost modeling for the manufacturing processes of viral vaccines. For this purpose, we typically use the BioSolve Process software package, developed and marketed by Biopharm Services.

The in silico modeling of production processes allows us to make informed decisions swiftly, saving considerable time and resources during the development phase and manufacturing of viral vaccines. This software seamlessly integrates up-to-date information from across the biopharmaceutical sector with the most recent industry-benchmarked price information for materials, consumables, and equipment.

Benefits of In Silico Cost Modeling

In silico cost modeling is a pivotal tool for comprehending the cost implications of technology and process choices. It enables us to design the most cost-effective process steps and select optimized technologies to support the process under development.

Through in silico modeling, risks can be substantially reduced by rapidly conducting multiple process comparison analyses and facility utilization assessments. We leverage the BioSolve Process software package to build business cases for innovative technologies or process options. We conduct comprehensive analyses of multiple process comparisons using the numerous analysis tools available before presenting our optimized, most cost-effective solution.

Beyond its use for novel processes, we’ve also successfully implemented in silico modeling to optimize existing biopharmaceutical manufacturing platforms. Our in silico modeling services provide our clients and us with the capacity to:

• Transition manufacturing scale from clinical to commercial
• Ascertain if a process will fit into preset facility designs
• Compare process choices and technologies from a cost perspective
• Examine various technology options to identify the optimal fit for achieving production targets
• Configure any viral vaccine process on upstream and downstream processing
• Design improved processes and screening technologies

With our in silico modeling expertise and software systems, we can significantly reduce manufacturing costs by understanding cost structures early in developing both non-platform and platform processes.