Process and product understanding is at the root of manufacturers’ efforts to produce safe and effective medicines. Researchers in our group at MIT are building novel first-principles and data analytics-based mathematical modeling tools for the manufacturing of biopharmaceuticals. To experimentally validate these modeling tools and to fully understand the impact of model choice on product quality, a fully instrumented and integrated continuous testbed for the manufacturing of monoclonal antibodies (mAbs) was constructed. The testbed consists of 4 parallel upstream systems including 4 perfusion devices, with one reactor assembly integrated with a fully continuous downstream system including Protein A chromatography, in-house designed viral inactivation, and ion exchange chromatography.
For process characterization in the upstream system, in-reactor probes for capacitance, optical density, DO, pH, and Raman spectra were integrated into the system. Additional analytics are made possible by the MAST autosampling system, which directs cell-containing samples to the Nova FLEX2 cell culture analyzer and cell-free perfusate samples to the Gilson liquid handler. There, samples can be navigated to an Agilent 1260 Bio-Inert HPLC for product titer and aggregation analyses or to an Agilent 6545XT LC-QTOF for at-line characterization of glycosylation profiles. Further characterization of product CQAs is performed in off-line workflows using the Agilent AssayMAP Bravo for sample preparation for subunit and peptide-level analyses. This presentation describes the instrumentation and discusses data collection and process integration within the testbed. The experimental data generated by this highly instrumented integrated continuous biomanufacturing testbed enable the evaluation, development, and validation of modeling methods and control strategies, and ultimately contribute to improved process understanding for better informed risk-based decisions during manufacturing campaigns.
3 Key Learning Objectives:
Information on assembly of a highly instrumented continuous manufacturing testbed
Process development on the testbed
Analysis of monoclonal antibody critical quality attributes using HPLC and LC-MS
The SARS-CoV-2 pandemic has challenged conventional paradigms to traditional vaccine development,
manufacturing, approval, and supply. A vision for all the elements necessary to a successful global
immunization program is essential, including considerations of the variety of vaccines platforms, such as
virus-like particles, subunit protein-based antigens, novel nucleic acid and vector-based vaccines, as well
as vaccines control strategies necessary to ensure continued supply of safe and effective product.
At the root of the development activities for each class of vaccine is scientifically sound characterization
of the vaccine, including vaccine components, such as lipid nanoparticles and adjuvants. Adherence to
principles associated with Quality by Design are important in an accelerated vaccine development
program, to guide process and analytical development studies. These are grounded in product
specifications and further supported by a risk-based life cycle change management program.
This session will focus on the work performed by several vaccine manufacturers to optimize their vaccine
candidates and develop commercial control strategies to facilitate the worldwide delivery of sufficient
safe and effective vaccines, which hold the promise of stemming the SARS-CoV-2 pandemic. Audience
interaction with these industry leaders will be supplemented with a panel discussion that includes global
regulators, who are currently evaluating these vaccines and hold special interests in vaccine control
Along with technical questions from participants, some topics that might expand the discussion towards
industry and regulatory coordination will include:
• Given the immediate need for a safe and effective vaccine, what do companies and regulators
perceive as the greatest technical hurdles to rapid development and adequate control of a vaccine?
• What are the special considerations, if any, associated with the development of a vaccine targeted
towards a pandemic pathogen?
• The interactions between companies and regulators, among regulators, and among companies
might differ for a pandemic development program versus a routine vaccine development program.
What technical expectations and practices, if any, should be different to help progress a pandemic
vaccine towards successful introduction to populations in need?
Diagnostic testing for active SARS-CoV-2 infection, testing for past infection, and analyses of potential vaccines are critical enablers of the treatment, understanding, and control of SARS-CoV-2. Diagnostic and other tests have also been key components of recent fights against viruses such as MERS-CoV, Zika, Influenza H7N9, and Ebola. Nucleic acid tests, serological tests, serum antibody tests, and neutralizing serum antibody tests are examples of population testing that have been employed for COVID-19 diagnostic testing. The current tests that have been given approval or authorization by Regulatory Authorities and are being used by vaccine manufacturers have been developed and reviewed in a timecritical fashion. There are many potential learnings regarding development, commercialization, supply, and regulation that can be taken from the SARS-CoV2-related activities of the past year and from work on previous pandemic diagnostics. These leanings can potentially be applied to both additional SARSCoV-2 undertakings and activities related to future pandemics; across industry, regulatory, and academic environments; and to diagnostics, vaccines, and therapeutics. At the same time, there remain many questions and points for further discussion. This session will discuss evolution and use of SARS-CoV-2 diagnostics and other tests from industry, regulatory, and patient care perspectives.
We welcome you to attend the WCBP 2021 Waters Technical Seminar, where we will cover several recent innovations in separations and mass detection, that will enable biopharmaceutical organizations to obtain better data more effectively. You will learn about the recent development of Waters High Performance Surface (HPS) technology, and the positive impacts that it can provide on bioseparations performance, efficiency, and robustness. We will also detail new capabilities of the BioAccord LC-MS System that continues to make LC-MS attribute-based analysis accessible to experts and non-MS experts alike across a biopharmaceutical organization.
Our guest speaker, Francesco Lanucara (Allergan, Liverpool, UK) will describe his group’s efforts to gain insights on Adeno-Associated Virus (AAV) capsid structures through multi-level LC-MS analysis. This complex gene therapy and vaccine delivery platform is subject to evolving regulatory expectations for both attribute characterization and monitoring. Allergan will be presenting their work with the BioAccord LC-MS System for attribute definition and transferring this knowledge into higher throughput monitoring assays, and potential automated attribute-based methods for AAV analysis within the highly regulated QC environment.
The pharmaceutical journey started with antibiotics and small molecules. This has shifted to large molecules such as antibodies and recently to more complex molecular entities such as next generation protein therapeutics, cell and gene therapies. As we know these newer molecules will revolutionize disease treatment, change the prognosis for many conditions, and help close the undruggable gap. The complexity of these molecules creates analytical challenges beginning at the clone selection stage through process development and commercial production, which has no doubt has already affected you and your team. Obviously COVID has accelerated growth in next-generation vaccines. This growth, and the urgent need for manufacturing of these new therapeutics and vaccines require faster and robust analytics for characterization and impurity analysis of both the molecules such as, bi- & tri-specifics, nucleic acids, oligonucleotides and mRNA, as well as their delivery systems such as viral vectors and lipid nanoparticles.
Over the last decades, capillary electrophoresis (CE), originally used for nucleic acid analysis and plasmid topology, has been successfully deployed to perform validated assays for product characterization and release testing of commercially available protein-based therapeutics. Likewise mass spectrometric based methods (LC-MS) are being increasingly implemented for both small and large molecule characterization and quantitation due to their high levels of precision and specificity. Presented here is a set of CE and LC-MS analytical workflows developed and optimized not just for characterization, quantification of various nucleic acid, viral vectors, LNP and next generation mAb variants, but also for process- and product-related impurities.