Peptide epitope fingerprints for YghJ, a vaccine antigen of interest for preventing urinary tract infections, were identified from Patient’s as well as immunized animals’ sera. The overall pattern of peptide epitopes recognized after immunization of rabbits and mini-pigs as well as infection in humans and mini-pigs is almost identical. In contrast, individual responses within each group apparently show an even larger deviation at epitope level. The cause for this is the acquired immune response due to repeated or continuous contact with the natural antigen. This pre-immunisation has a large influence on the vaccine’s effects.

The rapid development of safe and effective vaccines is a key requirement to conquering any pandemic. Recombinant proteins remain amongst the most reliable and safest approaches but face challenges in design, manufacture, structural characterization, analytical analysis, and delivery. The examples of the 2009 influenza pandemic and 2019 COVOD-19 pandemic highlight the many challenges faced when compressing development of a novel pandemic vaccine into months rather than decades.

This presentation will focus on providing an overview of high-throughput analytical characterization and formulation development considerations for new vaccine candidates targeted for use in low- and middle-income countries (LMICs). Case studies with live-virus vaccine candidates will highlight implementing state-of-the-art selective and stability-indicating assays to enable expeditious formulation development to enhance vaccine access in LMICs.

This talk will include analytical strategies for quantitative evaluation of critical quality attributes (CQAs) of mRNA vaccine drug substances (DS) and drug products (DP) with a view to speedy batch release of high quality vaccines. In-process measurements of CQAs can drive optimization of critical process parameters and help development of an optimized process that will yield high quality products. Strategies for addressing the challenges with evaluating integrity of each mRNA construct and potency of each antigen in a multivalent mRNA vaccine will be discussed.

Appropriate quality assessment is an important element of vaccine development and manufacturing. The USP-NF contains general information and best practices on the quality assessment of vaccines. This presentation will cover General Chapters in the USP-NF that support vaccines and new efforts to develop additional tools for vaccine quality assessment.

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.

The regulatory approval of ONPATTRO, a lipid nanoparticle-based short-interfering RNA drug for the treatment of polyneuropathies induced by hereditary transthyretin amyloidosis, paved the way for the clinical development of many nucleic acid-based therapies including the approved mRNA vaccines, enabled by nanoparticle delivery. The chemistry and biology principles behind the discovery, development and targeting based on LNP technology will be discussed.

Messenger RNA (mRNA) has great potential as a therapeutic or vaccine agent. We generated cationic nanolipoprotein particles (c-NLPs) to complex with traditional non-amplifying mRNA or self-amplifying mRNA (SAM) for in vivo delivery, evaluating protein expression relative to a lipid nanoparticle (LNP)-based delivery vector over 22 days. Furthermore, SAM formulated with c-NLPs or LNPs induced robust immune response against a variety of viral and bacterial antigens.

Adenovirus vectors have become an important class of vaccines with the recent approval of Ebola and COVID-19 products. In-process quality attribute data collected during Adenovirus vector manufacturing has focused on particle concentration and infectivity ratios (based on viral genome: cell-based infectivity), and data suggests only a fraction of viral particles present in the final vaccine product are efficacious. Here we examine the product heterogeneity of two Adenovirus viral vectors, Chimpanzee adenovirus (ChAdOx1) and Human adenovirus Type 5 using TEM and MS.

Non-woven material represents the next-generation of chromatography material with special features for bionanoparticles separation. The architecture of this bed allows the processing of large particles and molecules without diffusional limitation. This is exemplified by separation of COVID-19 and influenza virus-like particles.

Recently, the development of gene editing technologies (CRISPR-Cas,TALENs, zinc finger nucleases (ZFNs)) have opened new opportunities to precisely edit the genome, target disease causing mutations, and potentially enable one-time cures of genetic diseases. However, these therapeutics must overcome numerous obstacles to be successful, including rapid in vivo degradation, poor uptake into target cells, required nuclear entry, and potential in vivo toxicity in healthy cells and tissues. In this talk, I will discuss our efforts towards the development of lipid and polymer-based nanoparticles that enable the delivery of nucleic acid therapeutics to target cells and tissues in vivo. Furthermore, I will describe new therapeutic strategies utilizing these nanoparticles to (i) reprogram immune cells for cancer immunotherapy applications, (ii) in utero mRNA delivery for treating disease before birth.

The rapid deployment of three COVID-19 vaccines authorized for use in the United States in 2020 and 2021 involved substantial firm-level collaboration to rapidly complete the multidimensional matrix of steps along the entire global value chain from discovery to commercialization. Key enabling factors included quickly establishing and expanding international supply chains meeting regulatory requirements and the vaccine owners’ rapid technology transfer to firms in developing and developed countries.