Glassy-State Stabilization of a Dominant Negative Inhibitor Anthrax Vaccine Containing Aluminum Hydroxide and Glycopyranoside Lipid A Adjuvants

During transport and storage, vaccines may be exposed to temperatures outside of the range recommended for storage, potentially causing efficacy losses. To better understand and prevent such losses, Dominant Negative Inhibitor (DNI), a recombinant protein antigen for a candidate vaccine against anthrax, was formulated as a liquid and as a glassy lyophilized powder with the adjuvants aluminum hydroxide and glycopyranoside lipid A (GLA). Freeze-thawing of the liquid vaccine caused the adjuvants to aggregate and decreased its immunogenicity in mice. Immunogenicity of liquid vaccines also decreased when stored at 40 °C for 8 weeks, as measured by decreases in neutralizing antibody titers in vaccinated mice. Concomitant with efficacy losses at elevated temperatures, changes in DNI structure were detected by fluorescence spectroscopy and increased deamidation was observed by capillary isoelectric focusing (cIEF) after only 1 week of storage of the liquid formulation at 40 °C. In contrast, upon lyophilization, no additional deamidation after 4 weeks at 40 °C and no detectable changes in DNI structure or reduction in immunogenicity after 16 weeks at 40 °C was observed. Vaccines containing aluminum hydroxide and GLA elicited higher immune responses than vaccines adjuvanted with only aluminum hydroxide, with more mice responding to a single dose

Shifting the Burden of Corporate Taxes Heterogeneity in Direct Wage Incidence

We contribute to the empirical literature on the effective incidence of corporate income taxation. We focus on the so-called direct incidence via the wage bargaining process. Building on the innovative framework of Arulampalam, Devereux and Maffini (2012), we analyze the importance of various dimensions of heterogeneity at the firm-level. In particular, we investigate the distinct effects of (i) firm size, (ii) level of profitability, and (iii) competition intensity across (iv) different economic sectors. Furthermore, we investigate the relative importance of the surrounding institutional setting. To this end, a firm-level within-country approach is pursued separately for two different economies, namely France and the United Kingdom, which can be regarded as polar cases with respect to the relevant features of the wage-setting process. However, in many respects, we find surprisingly similar results for both countries. Thereby, this paper also adds to the literature by providing new insights on the degree to which results from previous single-country studies can possibly be generalized.Die effektive Inzidenz der Körperschaftsteuer ist ein theoretisch wie empirisch kontroverses Thema. Das Paper leistet einen Beitrag zur empirischen Literatur und fokussiert dabei auf die sogenannte direkte Inzidenz über den Lohnverhandlungskanal. Aufbauend auf dem innovativen Ansatz von Arulampalam, Devereux und Maffini (2012) wird die Bedeutung von verschiedenen Dimensionen der Heterogenität auf der Firmenebene analysiert. Konkret wird erforscht, welchen Einfluss (i) Firmengröße, (ii) Profitabilität und (iii) Wettbewerbsintensität in (iv) verschiedenen Branchen auf das Ausmaß ausüben, in dem die Last der Körperschaftsteuer auf die Löhne der Beschäftigten überwälzt wird. Darüber hinaus wird untersucht, welche Relevanz dabei den institutionellen Gegebenheiten der Lohnverhandlungen zukommt. Dazu werden alle Analysen separat für Frankreich und das Vereinigte Königreich vollzogen, deren Volkswirtschaften mit Blick auf Arbeitsmarktinstitutionen als polare Fälle gelten können. Die jeweiligen Ergebnisse für die beiden Länder sind sich jedoch qualitativ wie quantitativ überraschend ähnlich. Insofern gibt die Studie auch einen Hinweis darauf, inwiefern die Ergebnisse aus auf einzelne Länder bezogene Studien zur effektiven Lohninzidenz der Körperschaftsteuer generalisiert werden können

Ultra Stable Glassy State Vaccines Containing Adjuvants

Vaccines often require a narrow temperature range for storage during the cold chain. Damage to vaccines can occur if the vaccines are frozen, or exposed to elevated temperatures which could ultimately lead to a loss in vaccine efficacy. Lyophilized vaccines allow for a wider range of storage temperatures without having vaccines experience a decrease in efficacy. By utilizing rapid freezing kinetics and high concentrations of the glass-forming excipient trehalose, the particle size distribution of aluminum hydroxide adjuvant particles was maintained during lyophilization and reconstitution. Lyophilized recombinant ricin toxin A, dominant negative inhibitor, and human papillomavirus vaccines were equally as immunogenic as their liquid counterparts. The lyophilized vaccines were able to remain stable without protein structural changes or a decrease in immunogenicity after storage at an elevated temperature of 40-50 ºC, where liquid vaccines exhibited alterations in protein antigen structure and decreased immunogenicity. The addition of the tolllike receptor agonist, glycopyranoside lipid A was able to increase antibody titers and the rate of seroconversion for the anthrax vaccines but failed to do so for the human papillomavirus vaccines, showing that the immune response may be antigen specific. Although, no commercially available vaccines are lyophilized in the presence of an aluminum salt adjuvant, the work presented in this thesis provide evidence that lyophilization can be used successfully with aluminum hydroxide and glycopyranoside lipid A adjuvants

Rational Design and In Vivo Characterization of mRNA-Encoded Broadly Neutralizing Antibody Combinations against HIV-1

Monoclonal antibodies have been used successfully as recombinant protein therapy; however, for HIV, multiple broadly neutralizing antibodies may be necessary. We used the mRNA-LNP platform for in vivo co-expression of 3 broadly neutralizing antibodies, PGDM1400, PGT121, and N6, directed against the HIV-1 envelope protein. mRNA-encoded HIV-1 antibodies were engineered as single-chain Fc (scFv-Fc) to overcome heavy- and light-chain mismatch. In vitro neutralization breadth and potency of the constructs were compared to their parental IgG form. We assessed the ability of these scFv-Fcs to be expressed individually and in combination in vivo, and neutralization and pharmacokinetics were compared to the corresponding full-length IgGs. Single-chain PGDM1400 and PGT121 exhibited neutralization potency comparable to parental IgG, achieving peak systemic concentrations ≥ 30.81 μg/mL in mice; full-length N6 IgG achieved a peak concentration of 974 μg/mL, but did not tolerate single-chain conversion. The mRNA combination encoding full-length N6 IgG and single-chain PGDM1400 and PGT121 was efficiently expressed in mice, achieving high systemic concentration and desired neutralization potency. Analysis of mice sera demonstrated each antibody contributed towards neutralization of multiple HIV-1 pseudoviruses. Together, these data show that the mRNA-LNP platform provides a promising approach for antibody-based HIV treatment and is well-suited for development of combination therapeutics

Optimization of Lipid Nanoparticles for Intramuscular Administration of mRNA Vaccines

mRNA vaccines have the potential to tackle many unmet medical needs that are unable to be addressed with conventional vaccine technologies. A potent and well-tolerated delivery technology is integral to fully realizing the potential of mRNA vaccines. Pre-clinical and clinical studies have demonstrated that mRNA delivered intramuscularly (IM) with first-generation lipid nanoparticles (LNPs) generates robust immune responses. Despite progress made over the past several years, there remains significant opportunity for improvement, as the most advanced LNPs were designed for intravenous (IV) delivery of siRNA to the liver. Here, we screened a panel of proprietary biodegradable ionizable lipids for both expression and immunogenicity in a rodent model when administered IM. A subset of compounds was selected and further evaluated for tolerability, immunogenicity, and expression in rodents and non-human primates (NHPs). A lead formulation was identified that yielded a robust immune response with improved tolerability. More importantly for vaccines, increased innate immune stimulation driven by LNPs does not equate to increased immunogenicity, illustrating that mRNA vaccine tolerability can be improved without affecting potency. Keywords: mRNA, vaccines, LNP, intramuscular, lipids, tolerability, immunogenicity, formulatio