Vaccines play an important role in protecting the health of society and have saved millions of lives. By stimulating, and instructing, the immune system they can prevent infectious disease and offer great hope for therapeutic vaccines that can treat diseases such as cancer, Alzheimer’s disease and viral infections such as HIV. The manufacture of vaccines is classically focused on the direct generation, and purification, of an antibody generating (antigen) substance derived from, or simulating, some component of the infectious organism. Next generation vaccines utilize different methods to genetically instruct the body’s cells to generate these antigens. The manufacturing processes differ depending on the type of vaccine but share common attributes, and often technology, that assure the quality of the final drug and accelerate the journey from discovery to clinical manufacture.
Nucleic Acid Vaccines
Together with recombinant vaccines, nucleic acid vaccines represent the fastest growing class and are particularly suited to rapid vaccine development in response to emerging viral threats. The genetic material may be DNA or mRNA and is encoded to produce the targeted antigen in-situ. Delivery of the genetic material to the target tissues can use several vectors including lipid nanoparticles (LNP), viral vectors and plasmid DNA, or the uptake enhanced using techniques such as electroporation.
mRNA Vaccines
Chemically-synthesized and encoded to directly generate specific disease antigens, this new class of vaccine can be quickly developed and scaled rapidly to make clinical material available for trials and commercialization. They also hold promise for the prevention and treatment of various cancers by targeting tumor-associated antigens. The synthetic nature of this class of vaccine simplifies the manufacturing challenges compared to more complex biological processes and is generally perceived as having a strong and predictable safety profile as a result. Despite these apparent advantages, challenges relating to the stabilization and delivery of the genetic material to the target cells exist, and may provide particular challenges throughout the generation of drug substance and the final formulation and filling processes.
Specifically, the use of lipid nanoparticles (LNP) is being investigated as a delivery mechanism, however, it is known that these complex formulations can be susceptible to the low throughput of some sterilizing-grade filters and carry increased risk of bacterial penetration without careful filter selection and validation.
Precision NanoSystems
Precision NanoSystems (PNI), an innovator in lipid nanoparticles, joins the Danaher Life Sciences platform as a strong complement to Pall Corporation and Cytiva. Together, we will further enable you in your development of genetic medicines and in advancing your science to improve the lives of patients. Lipid nanoparticles are a key reagent for making mRNA therapeutics and other genetic medicines. The PNI Genetic Medicine Toolkit contains proprietary lipid nanoparticle and microfluidic manufacturing platforms. Learn more
Purification
Ultrafiltration membranes used during diafiltration and product concentration stages may be applied for size-based purification.
Anion exchange membrane chromatography may be applied for endotoxin control and change-based contaminant removal.
Filtration
Complex formulations, can create unique filtration challenges and careful filter selection is required to maintain high yields.
High-throughput capsules reduce filter size-related product loses and maintain high yields.
Fluid Handling
Maintaining the cleanliness of manufacturing processes is a central element of manufacturing in accordance with cGMP. The use of single-use systems can vary from the collection and transfer of process components in bioprocessing bags to complete integrated and automated single-use manufacturing processes. These support closed manufacturing processes that assist with multi-product manufacturing to deliver cost effective, responsive and agile processes.
DNA Vaccines – Plasmid Vectors
Vaccines using plasmid DNA to transfect cells cause the intracellular expression of one or several antigens relating to target pathogens or cancerous cells. While the plasmid may code for different changes, these are similar to some stages during the production of viral vectors used for gene therapy applications. The scale will vary depending upon the target disease however the production processes are essentially the same.
Single-Use Processing
From media preparation to cell culture and purification, the use of single-use systems and bioreactors, coupled by sterile connectors and disconnectors, helps maintain the safety at all stages of the vaccine production process, final formulation and filling. These intrinsically closed processing solutions also support rapid turnaround of different processes in multi-product facilities.
DNA Vaccines – Viral Vectors
Viral vector DNA vaccines use harmless viruses such as poxvirus and adeno-associated virus (AAV) to deliver eingineered genetic instructions to the target tissue. These sequences are encoded to cause these tissues to produce the desired anitgens which, in turn, then stimulate the immune system to generate antibodies.
These processes are shared with gene therapy applications and leverage the same technology, especially supporting industrial scale solutions for the adherent and suspension cell culture that multiply these viruses. When coupled with platform purification processes, thsese vaccines can be developed and industrialized rapidly.
Single-Use Processing
From media preparation to cell culture and purification, the use of single-use systems and bioreactors, coupled by sterile connectors and disconnectors, helps maintain the safety at all stages of the vaccine production process, final formulations and filling.
Recombinant Virus Vaccines
Viruses, engineered to look like a wild virus strain to a body’s immune system, but without the undesirable effects, hold much promise as platform vaccines. The manufacturing processes share similar technologies with both gene therapy and monoclonal antibody production depending upon the producer cell-line.
Upstream
Single-use bioreactors to support both suspension and adherent cell cultures
Sterilizing grade gas filtration
Sterilizing grade liquid filtration for media filtration and sterile additions
Harvest and Clarification
Maintaining a high virus yield while separating the virus from the host cells requires careful optimization. The use of single-use depth filtration platforms simplify the clarification and the choice of the right grades will ensure recovery is optimized.
Media Preparation
Solutions for buffer and media preparation to support upstream processing include mixing applications, storage and transport. Additional operations such as sterile filtration can be integrated with mixing technologies to create robust automated solutions.
Purification
Concentration and diafiltration for size-based separation of contaminant using ultrafiltration cassettes.
Anion exchange membrane chromatography simplifies the removal of endotoxin and change-based contaminant removal such as host cell DNA, proteins and endotoxin. It may also be applied for bind-elute operations to purify and concentrate virus without high shear ultrafiltration.
For processes including mammalian cell cultures viral safety is an important consideration and can be managed with orthogonal controls such as inactivation and virus filtration.
Recombinant Vaccines
Yeast, bacterial, mammalian and insect cell lines may be used depending on the complexity of the antigen being produced and several may be combined (conjugated) to elicit the desired immune response. When using mammalian cell lines, these processes share the upstream challenges of monoclonal antibody production and leverage the range of cost-effective solutions available for the development and manufacture of these vaccines.
Upstream
Maintaining a monoseptic environment is essential for a successful vaccine production process. This can be supported with the use of single-use technology integrating filtration and single-use bioreactors.
Sterilizing grade gas filtration
Sterilizing grade liquid filtration for media filtration and sterile additions
Harvest and Clarification
Partnered with centrifugation or used on its own, the removal of whole cells and cellar debris can be easily achieved using single-use depth filtration systems optimized for throughput and yield.
Stax single-use depth filtration platforms support the simple clarification of post-centrifugation and low turbidity cell cultures.
For mammalian and insect cell cultures the Stax mAx clarification platform provides a simple, scalable solution even for high turbidity harvest.
For challenging bacterial lysates the PallSep Biotech MF system provides a robust solution for low volume <500 L processes.
Media Preparation
Solutions for buffer and media preparation to support upstream processing include mixing applications, storage and transport. Additional operations such as sterile filtration can be integrated with mixing technologies to create robust automated solutions.
Purification
Concentration and diafiltration for size-based separation of contaminant using Delta ultrafiltration cassettes.
Mustang XT Anion exchange membrane chromatography for endotoxin control and change-based contaminant removal such as hot cell DNA, proteins and endotoxin.
For processes including mammalian cell cultures viral safety is an important consideration and can be managed with orthogonal controls such as inactivation and virus filtration.
Fluid Handling
Maintaining the cleanliness of manufacturing processes is a central element of manufacturing in accordance with cGMP. The use of single-use systems can include unit operations such as the collection and transfer of process components in bioprocessing bags to complete integrated and automated single-use manufacturing processes. These support closed manufacturing processes that assist with the rapid turnaround and multi-product manufacturing to deliver responsive and agile processes.
Bacterial Toxoid Protein Vaccines
Using inactivated bacterial toxins (toxoids) as the antigen, these vaccines protect against certain bacterial threats such as diphtheria and tetanus.
Upstream
Maintaining a monoseptic environment is essential for a successful vaccine production process. This can be supported with the use of single-use technology and maintained with filtration.
Sterilizing grade gas filtration
Sterilizing grade liquid filtration for media filtration and sterile additions
Harvest and Clarification
Partnered with centrifugation or used on its own, the removal of whole cells and cellar debris can be easily achieved using single-use depth filtration systems optimized for throughput and yield.
Stax single-use depth filtration (e.g grade K100P)
Media Preparation
Mixing solutions for buffer and media preparation to support upstream processing and formulation
Purification
Concentration and diafiltration can achieve the size-based separation of contaminants using ultrafiltration cassettes.
Anion exchange membrane chromatography may be used for endotoxin control and the removal of contaminants through charge-based interactions.
Fluid Handling
Kleenpak® Presto sterile connectors and Kleenpak sterile disconnectors control risks associated with the connection and disconnection of fluid paths to safeguard operators, quality and simplify processing when using single-use manufacturing technology. Such solutions are perfect for the aseptic formulation of multivalent vaccines where sterile filtration may not be possible after the addition of some adjuvants.
Live Attenuated Vaccines
First generation vaccines, include the majority of influenza vaccines and use attenuated or inactivated virus. In the case of influenza vaccines these are historically produced from the inoculation of embryonic chicken eggs. Recent developments using mammalian cell-based cultures such as human embryonic kidney (HEK)-293 cells or Manin-Darby canine kidney (MDCK) cells have the potential to improve the effectiveness of the vaccine and permit greater process flexibility.
Maintaining sterility throughout a series of large-scale vaccine manufacturing and formulation steps requires careful process design. The need for strict controls increases with the addition of some adjuvants which make the addition of sterilizing grade filtration impractical due to the removal of these adjuvants by the filtration process. To meet these challenges, aseptic conditions need to be maintained throughout the process and can be simplified through the use of single-use closed processes.
Harvest and Clarification
Solutions vary depending upon the cell culture process. For cell-based vaccines, these also depend on the cell-line however single-use depth filtration is a strong
For processes such as egg-based cultures the use of polymeric depth filters work well in conjunction with centrifugation to remove solid contaminants ready for further purification and formulation.
Fluid Handling
Kleenpak® Presto sterile connectors and Kleenpak sterile disconnectors help to maintain the sterility during the connection and disconnection of fluid paths throughout the process. When combined with bioprocessing bags and tubing transfer sets these single-use fluid flowpaths can be used to combine multiple strains of vaccine to simplify the processing while safeguarding quality.