Process Development

Executive Summary

• In Mabion we perfectly understand that the key to global success in bio-tech industry is a robust, reproducible and well-controlled production process. This is why, we develop and constantly improve know-how that allows us to provide professional and cost-effective services in the field of process and analytical development. Expertise of our team combined with state-of-the-art equipment and well-controlled quality management system enables us to implement customized solutions for projects of various complexity levels.
• Process development at Mabion covers the complex pathway from derivation of a stable cell line, through small-scale protein production and purification, to the design of full-scale manufacturing process. We have experience with both mammalian and insect cell lines, production of monoclonal antibodies and other recombinant proteins, but our approach to process development is always customized to the particular product and its unique characteristics.
• Our team strongly believes that the implementation of the Quality by Design approach is the best way to guarantee the successful project outcome. That’s why we start designing fit-for-purpose analytical methods right from the earliest project phases, and then use them to control the successive phases of process development, and ultimately the process itself. This gives us a thorough understanding of relationship between the process and the product, which leads to better control, more conscious development and scale-up as well as safe commercial production in the future.
• Our process development services include the design of upstream, downstream and up-scale for processes, including clonal evaluation and selection, culture media testing, protein purification and final product characterization. The key asset of our company is a strong analytical background, with multiple different methods available for use in process development and quality control.

Quality by Design

Quality by design (QbD) framework is essential to improve and enhance robustness of the biologics manufacturing process and designing of analytical methods. Impact of QbD on the pharmaceutical industry has been reviewed in many publications, which stress the capabilities of this approach to increase the product quality, efficacy and safety. The concept of QbD can be applied to analytical methods as well, where the focus is on designing and developing methods that are robust, reliable, and provide accurate and precise results. Mabion implements QbD throughout the product and process development achieving outstanding productivity and quality of the manufactured products.
QbD approach has a particularly high magnitude of benefit in the stage of product and process development and characterization, technology transfer, product and process monitoring and regulatory filings. The benefits include:

• For process development and characterization: operation at optimal conditions, greater flexibility and control, and increased knowledge on the molecule itself;
• For technology transfer: reduction in risk of failure or delays;
• For process monitoring and regulatory filings: quality assurance during product lifecycle.

Clone Evaluation and Selection

In the process of derivation of cell lines producing therapeutic proteins, it is extremely important that the line is monoclonal, stable, highly productive and that the resulting product has a quality profile that is consistent with expectations.

Experience and knowledge are crucial in this area as is the right equipment. Our scientists always use high-throughput systems to complete the process of clonal selection of cells from heterogeneous populations.

Thanks to the combination of knowledge and technology, we are able to select the clones with the highest productivity, which is one of the most important attributes for industrial applications.

Upstream Process

Our offer includes the full design of the upstream manufacturing process, from clone selection through optimization of the culture conditions and ending with the efficient expression of the desired product.
The most important services provided by Mabion are:

• Derivation of the stable/transient cell line
• Clone stability testing
• Production and testing of research cell banks
• Optimization of cell culture at lab scale (from 10 ml to 10L)
• Small-scale process characterization and scale-down validation studies

Culture Media Testing

During the development of culture conditions for antibody-producing cells, testing the levels of nutrients in the culture media and metabolites secreted by the cells during culture plays an extremely important role.

For this purpose, we use fully automated, high-throughput equipment that allows 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. Thanks to that, 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 upstream process, our team of specialist can take care of providing the best solutions for downstream processing. In our work, we always adopt the holistic approach, viewing the entire manufacturing process as a complex interplay of many factors functioning at different stages of production. This approach guarantees the development of reproducible, reliable and efficient process that is able to pass all regulatory expectations.

Among our services related to the downstream process development one can find:

• Harvesting (through depth-filtration and centrifugation)
• Viral inactivation and clearance
• Protein purification using various chromatography techniques (from 1mL to 100mL)
• Protein concentration (normal and tangential flow filtration)
• Small-scale process characterization and scale-down validation studies

Process Scale-Up

Our multidisciplinary team performs scale-up from the laboratory level, through the scale of 500L (if required), to the fully commercial scale of 2000-5000L. The entire process is done with thoughtful consideration of the EMA and FDA guidelines (incl. FDA 21 CFR Part 11 standards) as well as the outcomes of our partner’s interactions with regulatory agencies.
Scale-up process performed by Mabion includes the following stages:

• Determination of acceptance criteria (PAR – Proven Acceptable Range, NOR – Normal Operating Range) for individual process parameters, determination of critical product quality attributes (CQAs), critical process parameters (CPPs) and key process parameters (KPPs).
• Process characterization and determination of the process space for individual stages of drug product manufacturing (SDM, OFAT, QbD).
• Testing the purity, quality and stability of active substances and finished products.

Protein Purification

In the development of purification process, we use preparative systems that have their counterparts on industrial scales. We use them to develop purification methods, as well as to optimize the existing ones.

For design of experiments (DoE) as well as their control we use advanced, specialized software that is tailored to our process equipment. Our attention to experimental work results in high performance of the processes we develop.

Thanks to the appropriate design and software, our devices ensure control over the process conditions, as well as their proper monitoring and data archiving. Additionally, we are able to simulate the future course of processes on a laboratory scale down models to the smallest detail. This reduces the time of process implementation as well as the cost of transfer.

Product Characterization

In order to prove that the obtained product has the appropriate quality characteristics, we perform a number of physicochemical and biological analyses. Together they allow for a comprehensive structural and functional characterization of protein products.

Our laboratories are equipped with cutting-edge analytical systems. Through their use, we are able to develop, optimize, qualify and validate high-quality analytical methods that meet all requirements of regulatory agencies. Analyses performed by Mabion include:

• Evaluation of critical protein quality attributes (e.g., glycosylation profile, aggregates) using modern high-performance liquid chromatographies (HPLC, UHPLC). 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).