Process Development

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• Based in the EU, Mabion partners with clients throughout the world and offershigh-quality, cGMP compliant manufacturing of biologic therapeutics in cleanroom grades D through A, at competitive price points.
• Our dedicated team of specialists understands that each molecule, whether a biosimilar or an originator, has its unique characteristics which require a customized approach to development, optimization and production. The combination of our staff’s extensive experience and knowledge together with cutting edge technologies enables us to efficiently design and develop streamlined manufacturing processes, resulting in outstanding product yields and quality within mutually agreed project timelines.
• With our highly trained team and efficient planning, we can produce multiple batches utilizing a campaign mode to ensure maximum efficiency. In line with modern biopharmaceutical manufacturing practices, our state-of-the-art facility utilizes single-use technologies, ensuring that your product is free from contamination and minimizing switchover time between different products.
• At Mabion we appreciate the uniqueness of each biologic molecule. For this reason, when designing a manufacturing process, we always pay close attention to the characteristics of the particular product, doing our best to apply the multi-year experience and knowledge to the new project. Let us build the quality of your process from the very start, ensuring smooth development pathway and successful regulatory approval.

Early Process Development

Our process development for Phase 1 clinical studies focuses on designing a robust and scalable process that lays the foundation for the success of your biologic product. We understand the critical importance of a reliable process at this early stage, as it impacts the speed and quality of your drug’s progression through the pre-clinical and clinical pipeline.

Our team begins by conducting a thorough analysis of your biologic molecule’s characteristics and desired therapeutic properties. Using this data, we design a custom development plan that includes cell line development, media optimization, and initial upstream and downstream processing steps. We leverage high-throughput screening and Design of Experiments (DoE) methodologies to identify the optimal conditions for cell growth, protein expression, and purification. Our process development services ensure that the process is not only suitable for Phase 1 studies but also establishes a strong foundation for later-stage development.

Clone evaluation and selection

The first step in developing a successful manufacturing process for biologics is selecting the optimal cell clone. It is critical to ensure that the line is monoclonal, stable, highly productive and yields a product with a consistent quality profile, aligning with the QTPP.

Our clone evaluation and selection process begins by creating multiple cell lines expressing your molecule. These clones are rigorously evaluated for several key attributes, including productivity, growth characteristics, genetic stability, and expression of the desired protein.

We use high-throughput screening techniques and statistical analysis to rapidly evaluate and rank clones, ensuring that the most robust, stable, and high-yield one is selected for further development. Due to early selection of the optimal clone, we can minimize variability, increase productivity, and establish a solid foundation for the next steps of the manufacturing process.

Upstream process

Upstream process focuses on maximizing the yield and quality of the biologic product produced by the selected clone. Our upstream process development services for Phase 1 clinical studies are designed to create a scalable, repeatable process that ensures consistent and efficient production of your biologic candidate.

At Mabion, we employ a data-driven approach to upstream development, starting with milliliter volumes to define optimal conditions for cell growth and protein expression, through scaling the process to 10L bioreactors (Orbital Shaking Reactor (OSR) and Stirred Tank Reactor (STR)) and ending with a technology transfer to the Manufacturing Department (refer to our DS manufacturing and DP manufacturing services). We assess key parameters such as temperature, pH, oxygen, and feeding strategies to establish the ideal environment for the biologic-producing cell line. Our scientists use advanced tools like Design of Experiments (DoE) and high-throughput systems to evaluate and optimize these parameters. This method allows us to refine the process, ensuring that it is scalable for larger bioreactors without compromising product quality.

Our focus on process consistency and robustness during upstream development ensures that you can seamlessly proceed from lab-scale to clinical manufacturing with confidence, meeting the stringent requirements of first-in-human studies.

Key aspects of our lab-scale cell culture optimization process include analyzing interactions of such parameters as:

Cell culture media

• The optimal media composition, which is selected by analyzing diverse characteristics of cell lines, processes, media components (incl. their interactions), and metabolic pathways (see Culture media testing services below for further information).
• We also explore feeding strategies, such as fed-batch and perfusion, to improve cell viability and product yield.

Supplementation

• Selection of appropriate supplements and their specific times of addition to the cell culture allow for achieving high cell productivity and expression of proteins with defined attributes. During the cell culture development, we monitor levels of nutrients and metabolites in the media, using a fully automated, high-throughput equipment capable of performing hundreds of tests within hours. This technology enables us to quickly determine the appropriate supplementation strategy that enhance culture efficiency and product quality.

Seeding density

• We optimize seeding density allowing for cells to reach their maximum density while maintaining highest production capacity.

Culture duration and harvest

• Our experts determine the optimal culture duration to ensure maximum target protein production while minimizing impurities such as host cell proteins (HCP).

Biphasic culture

• In a biphasic culture, cells are initially grown to a high density, followed by a production phase in which cell growth is slowed or halted to boost protein production.
• Various approaches can be used to initiate transition between these two phases, of which the most common is switching temperature to mild hypothermia. Other strategies involve adding special reagents such as sodium butyrate (NaBu) or dimethylsulfoxide (DMSO), which are also capable of decelerating cell growth and extending the production period.

Culture media testing

We offer extensive culture media testing and optimization services to identify the best formulation for your specific biologic product and cell line.

For testing the nutrient levels in culture media, we use a fully automated, high-throughput equipment allowing us to perform several hundred tests within a few hours. The analysis of the content of culture fluid components is performed on the basis of enzymatic reactions and colorimetric measurements.

Based on the above, we can choose appropriate conditions of cell supplementation to increase the efficiency of the culture and improve the quality of the obtained product. We also monitor the content of metabolites that may have a negative impact on cell growth and viability.

Downstream process

Following the development of the upstream process, our experts start working on best strategies for downstream processing to ensure that your product meets the required purity, quality and regulatory standards to enter the clinical development stage or commercial use. We always aim to adopt a holistic approach, viewing the entire manufacturing process as a complex interplay of numerous factors interlacing at different stages of production. This approach guarantees the development of a repeatable, reliable and efficient process, capable of removing all impurities while maintaining the activity and integrity of your biologic molecule.

Our services include the development and optimization of the following process steps:

• Harvesting (through depth-filtration or centrifugation)
• Viral inactivation and clearance
• Protein purification using various chromatography techniques (affinity, CEX, AEX and other)
• Protein concentration and buffer exchange (normal and tangential flow filtration)

Similarly to other services, downstream process (“DSP”) development is driven by the QbD approach, with particular emphasis on the critical quality attributes of the product, which are monitored throughout the process development, thanks to our broad, in-house analytical panel (refer to our page on Analytics services). We begin by screening commercially available resins with the aim to select the most suitable ones for process optimization in the microliter scale (approx. 1mL). Larger columns are then packed and tested in-house by our scientists preparing the process for scale-up and to conduct further studies such as process characteristics or resin lifetime studies at the laboratory scale.

To ensure a flawless scale-up, all equipment used in early development phase has its equivalent in the targeted large scale manufacturing process. Importantly, we arrange continuous process knowledge transfer as your product evolves from lab scale to commercial scale production.

Process scale-up

Once your biologic product successfully completes early-stage development, the focus shifts to scaling up the manufacturing process to meet the demands of larger Phase 2/3 clinical studies and future commercial production.

Our multidisciplinary team specializes in process scale-up, from the laboratory level through intermediate scale of 500L (if needed) until the full commercial production scales of 2000L-4000L (soon to be upgraded to 8000L). The provided services include optimizing cell culture conditions and product yield, refining the purification processes, and implementing process controls that ensure repeatability and compliance with regulatory standards. We also perform scale-down modeling to mitigate potential challenges that could arise during commercial-scale production. The entire process is carried out in strict adherence to both EMA and FDA guidelines (incl. FDA 21 CFR Part 11 standards). We provide regulatory support throughout the scale-up process, including the preparation of documents for regulatory submissions, such as IND, NDA, or BLA applications.

Mabion’s facilities and equipment are designed for rapid and efficient scalability to smoothly transition your product from the lab to the clinical manufacturing environment and later to commercial manufacturing, while maintaining product quality and consistency.

To ensure flawless scale-up, all equipment used in early development phase has its equivalent in the target manufacturing process. Importantly, we arrange continuous process knowledge transfer as your product evolves from lab scale to mass production.

Our scale-up process consists of the following elements:

Development of control strategy

• We establish a strategy to manage process parameters within acceptable ranges and provide a preliminary determination of Proven Acceptable Range (PAR) and Normal Operating Range (NOR) for individual parameters, based on process development data.

CQAs, CPPs and KPPs determination

• We meticulously identify and define the CQAs, CPPs and KPPs parameters to ensure consistent product quality during scale-up.

Process optimization

• Before scaling, we conduct a thorough review and optimization of your existing process. This includes refining critical process parameters (CPPs) and identifying opportunities to enhance efficiency, yield, and product quality.

Pilot-scale production

• We bridge the gap between the laboratory and commercial production with pilot-scale runs. They help us validate the process scalability and identify potential challenges in a controlled environment.

Technology transfer

• Our robust technology transfer protocols ensure a smooth transition from development to manufacturing. We work closely with your team to transfer process knowledge, equipment specifications, and quality control methods, ensuring that every aspect is fully understood and replicated at scale.

Process validation

• We conduct rigorous validation studies to confirm that the product after process scale-up meets all predefined quality attributes. These studies include the production of multiple validation batches under cGMP.

Manufacturing scale-up

• Once validated, we scale up the process to full commercial production. Our advanced facilities are equipped to handle high-volume manufacturing while maintaining the highest quality standards and regulatory compliance.

Ongoing optimization

• We continue to monitor and optimize the process after completing scale-up to ensure it remains efficient and reliable. Your product will benefit from the latest advancements in manufacturing technology as a result of our commitment to continuous improvement.

Process optimization

As your biologic progresses to late-stage clinical studies (Phase 2, Phase 3 and sometimes Phase 4), process optimization becomes crucial to maximize yield, minimize costs, and ensure consistent product quality and commercial success. Our services are designed to optimize every aspect of your manufacturing process to meet the rigorous demands of large-scale production and regulatory compliance.

Our approach includes a thorough analysis of the current manufacturing process to identify potential areas for improvement. We utilize advanced analytical techniques, including real-time monitoring, to gain insights into process performance and product quality.

Our team works closely with our clients to implement changes that enhance process robustness, increase stability, and improve overall efficiency. We also ensure that all process changes are thoroughly documented and validated in line with regulatory requirements, preparing your product for large-scale trials and commercialization.

A key focus of our late-phase optimization is the development of a Scale Down Model (SDM), a critical tool that allows us to simulate large-scale manufacturing conditions at a smaller scale. This model helps establish the Proven Acceptable Range (PAR) and Normal Operating Range (NOR) for critical process parameters, providing a clear framework for robust process control.

One of the cornerstone aspects of our approach is the definition of a design space, a central element of Quality by Design (QbD) approach in biologics development. By clearly defining the design space, we provide flexibility within predefined boundaries, enabling us to make necessary process adjustments without compromising product quality or compliance. During this phase, we conduct extensive experiments combined with advanced statistical analyses to gain insights into how various process parameters interact and affect product yield, quality, and purity. With this comprehensive understanding, Mabion ensures that your manufacturing process is fully optimized, repeatable, and capable of scaling to meet commercial demands with confidence.

Process characterization

Understanding the intricacies of the manufacturing process and the key properties of the manufactured molecule is essential for ensuring product quality, safety, and efficacy. Our process characterization studies provide a deep dive into the crucial parameters and attributes that impact your biologic’s production and quality. These studies are vital at the late stage of development and play a key role in regulatory submissions and process validation.

Our process characterization services include a systematic evaluation of the process parameters and their impact on critical quality attributes (CQAs) of the manufactured biologic product. We employ robust experimental designs, such as DoE, to identify and establish the process design space. This includes determining the acceptable ranges for process parameters and understanding their interactions. Using advanced analytical tools and statistical methods we successively build a thorough understanding of the process, ensuring that it is well-characterized, controlled, and capable of consistently producing high-quality biologics.

In order to confirm that the properties and quality parameters of the obtained drug substance are in line with its specification, we perform a panel of physicochemical and biological tests. Our in-house laboratories are equipped with cutting-edge analytical devices, which we use for development, optimization, qualification and validation of high-quality methods that adhere to all relevant EU and US regulations. Together they allow for a comprehensive structural and functional characterization of the manufactured protein, ensuring its sufficient potency and safety for the patients.

Below is a list of Mabion’s in-house analytical methods:

• Evaluation of critical protein quality attributes (e.g., glycosylation profile, aggregates) using modern high-performance liquid chromatography systems (HPLC, UHPLC). The UHPLC system is equipped with three types of detectors: UV-VIS, fluorescence and corona discharge. One of the systems is also coupled to MS.
• Determination of amino acid sequence and post-translational modifications (using MS and other methods)
• Viral inactivation and clearance
• Determination of protein purity (using capillary electrophoresis with modular UV-VIS and LIF detectors)
• Measurement of interactions between molecules, incl. affinity and association/dissociation constants (using state-of-art SPR technology)

In addition to using our own arsenal of analytical methods, we proud ourselves to be experts in method transfers. During the challenging times during the COVID-19 pandemic we managed to perform a rapid and flawless transfer of an analytical method panel for a novel protein vaccine developed by Novavax, demonstrating our value as a trustworthy CDMO partner (see recommendation letter from Novavax). Our knowledgeable team can seamlessly transfer any protein characterization method to Mabion’s laboratories, while fully maintaining its accuracy, precision, reliability and efficiency. This smooth and seamless introduction of analytical techniques plays a pivotal role in ensuring the appropriate product quality and regulatory compliance.

Our QC and regulatory staff will prepare all necessary documents related to the process development and characterization, set-up of analytical methods, method transfer and other steps of the manufacturing process. You will receive the complete documents package required for filing the marketing authorization application and demonstrating compliance.

For more information on product characterization, refer to our page on Analytics services.