C1-cells have been shown to express antigens at > 100 times higher, in less time, without the need to remove viruses compared to baculovirus cells. C1-cells can express multicomponent vaccines, such as VLPs and ferritin nanoparticles. C1 strains have been shown to express more than 2.5 g/L (in 4-5 days), using low-cost chemically defined media in stainless-steel or single use microbial E. coli bioreactors. C1 cells lines have been used to stably express a growing number of antigens such as SARS-CoV-2 (i) Full Spike and (ii) RBD’s; H1, H5 & H7 Hemagglutinin; Neuraminidase, West Nile, Mumps, Rabies, Rift Valley Fever, etc.

• Recombinant proteins remain amongst the most reliable, low cost and safe approaches to pandemic vaccine delivery
• Protein vaccines face challenges in design, manufacture, structural characterisation, and scale up
• Adjuvants are critical to protein vaccine success
• Artificial intelligence tools can dramatically accelerate the process of vaccine design and optimisation?
• Now is the time to closely examine all aspects of pandemic vaccine design, manufacture, and supply chains to see how to do it better next time
  

My laboratory has developed a novel vaccine platform, virus-like particles (VLPs), formed with the structural proteins of Newcastle disease virus (NDV), an avian paramyxovirus. Using our platform, we have constructed VLPs containing the respiratory syncytial virus (RSV) F and G glycoproteins. Our preclinical studies have shown that these VLPs are excellent vaccine candidates for protection from RSV infection of neonates by maternal immunization and the protection of the elderly.

Developability assessments are key in making decisions on how, and indeed if, a therapeutic or vaccines candidate can move from bench to clinical trial.  Yet, these assessments are only as good as we design them.  It is crucial to ensure the assessment performed in any one organization includes more than a panel of analyses appropriate to the program molecules.  The assessment design and implementation must also take into account program capabilities and goals, limitations, and risk tolerance.  In this presentation we will discuss the elements of development assessments, their alignment with pre-formulation programs, and successful incorporation into early stage development programs.

Vaccine production processes are continuously optimized to produce high and pure viral titers, but analytical methods to determine the infectious titer in bulk, still lag. TCID50 and plaque assay are still gold-standard virus potency assays and rely on the induction of a cytopathogenic effect (CPE) as an endpoint for evaluation. These morphological changes are usually only visible after 1-2 weeks of infection and can be replaced by cellular events induced early after virus infection like a pro-inflammatory innate immune response in the form of secreted cytokines.

The development and production of vaccines & cell and gene therapies can be challenging and time-consuming due to the complex nature of the raw materials, process and final product.  Many current analytical methods, especially for viral vectors and vaccines, face challenges in terms of speed, reproducibility, and resource requirements. Here we present the application of laser force cytology as a real-time PAT to rapidly characterize viral infectivity and other critical quality attributes (CQAs) within minutes to inform and optimize the production process as well as its application for offline release and potency assays used to ensure product quality and consistency.

Sanofi’s influenza vaccine is the only recombinant influenza vaccine approved by the US Food and Drug Administration (FDA) and by the European Medicines Agency. Successful commercialization of influenza vaccines is a key growth driver for Sanofi Pasteur to increase the yield, decrease the cost per dose, and increase Manufacturing (MFG) capacity. We developed a new Fed-Batch process for production of recombinant HAs that results in 80% increase in rHA yield compared to the current production process.

• Continuous manufacturing
• Reducing costs for mRNA synthesis 
• Formulations for thermostability 
  

We have found that bacterial extracellular nano-vesicles (bEVs) may be useful as a strong adjuvant and displays the M2e antigen as an ion channel, thereby perhaps configuring the antigen in its native ion channel structure on a viral-infected host cell. Since bEVs co-express a lipopolysaccharide (LPS), we have improved the safety and tolerance of the nano-vesicles by creating a genetically-modified cell line that does not induce high levels of pyrogenicity. We found that a bEV-expressing M2e protects against fatal H5N1 infection in ferrets as well as mice at nano-gram doses with two immunizations.

• A lot of choices of membrane absorbers and fiber materials for virus purification compared to the past 
•  What is the selection criterion for a membrane adsorber and  fiber material 
•  The ideal separation process for a viral vaccine
  

Polysaccharide Conjugate vaccines are the most successful preventatives against bacterial disease and are composed of defined polysaccharides individually conjugated to carrier protein. This presentation will provide an update on standards and tools to support vaccine quality from raw materials through release testing. The recent revisions to General Chapter <198>, focused on NMR for bacterial polysaccharide for identity, and reference materials to support testing of quality attributes will be discussed.

The focus of the presentation is evaluation of wNMR as an emerging noninvasive analytical technology to characterize aluminum-adjuvanted vaccine formulations. In this work, wNMR and optical techniques such as laser diffraction and laser scattering were used to characterize vaccine formulations containing different antigen loads adsorbed onto AlPO4 adjuvant microparticles, including the fully dispersed state and the sedimentation process. The results of the study will be discussed.

We present a quantitative single-particle imaging platform that enables simultaneous measurements of the size, mRNA-payload, and dynamic properties of vaccines in cell-like conditions. We investigate the dependence of mRNA-lipid-nanoparticle structure and fusion dynamics on formulation, using commercially available formulations as a starting point. These measurements are made on confined, freely diffusing particles, and during reagent-exchange such as in response to solution pH, in order to emulate intracellular dynamics in a controlled setting. Over the long term and in collaboration with health scientists, we propose to correlate multi-scale data sets including single-particle measurements made in vitro as well as in cells and tissues, with clinical results, to create a throughline of understanding of vaccine effectiveness from the microscopic to clinical scale, to enable and optimize their rational design and engineering.

Light scattering techniques (DLS) for protein size characterization takes an ensemble-based approach to particle sizing. Although being a gold standard, this approach may not accurately distinguish particle sizes in a multimodal sample where the diameter of particles is similar or when particle size distributions are broad. Additionally, concentration of large particles can be overestimated thereby skewing the particle size distribution of the sample. On the other hand, single particle analysis approach involves tunable resistive pulse sensing (TRPS) technology which measures the size of individual particles passing through a nanopore and claims to provide more accurate particle size distribution data within a sample. Particle concentration analysis is also based on single-particle measurements thereby ensuring highly accurate calculations for each size band compared to the ensemble-based approach. The present study performs the comparative analysis of size distribution of vaccine antigens to evaluate the performance of these methods.

Despite the recent groundbreaking advancements in mRNA vaccine development, analytical methods have not evolved at the same rate, leaving a significant need for highly sensitive and rapid purity assessment methods. Here, we propose a dual dynamic staining high-throughput microfluidic electrophoresis analytical method for the detection and characterization of dsRNA contaminants in mRNA vaccines. With an mRNA maximum loading capacity of 13 ng/μL, we can detect dsRNA contaminants as low as 0.1-0.6% of the total concentration.