Live attenuated varicella live virus vaccine (Vabiotech)

The effectiveness of three Porcine Reproductive and Respiratory Syndrome (PRRS) Modified Live Virus (MLV) vaccines against PRRSV viraemia and nasal shedding following experimental challenge was compared. The study comprised a negative control (T01), and three treatment groups (T02, T03 and T04) each vaccinated with a single dose of a commercial PRRS MLV vaccine, given in accordance with the vaccine's Summary of Product Characteristics (SPC). Pigs aged 21 days were vaccinated (day 0), challenged intranasally (day 28) with heterologous PRRSV-1-1 strain Olot/91, then monitored for PRRSV viraemia and nasal shedding for 12 days. After challenge, pigs were viraemic on fewer days in group T04 (0.67) than groups T01 (0.91), T02 (0.81) and T03 (0.97) (P < 0.0296). From day 34, inclusive, serum PRRSV titres were lower in group T04 than negative controls (P ≤ 0.0001) and groups T02 and T03 (P ≤ 0.0047); serum PRRSV titre Area Under the Curve (AUC) for group T04 (42.34) was lower than in T01 (65.49), T02 (60.67) and T03 (67.38) (P < 0.0100); pigs exhibited nasal shedding on fewer days in group T04 (0.40) than T01 (0.78), T02 (0.64) and T03 (0.56) (P < 0.0101); and nasal shedding AUC for group T04 (8.52) was lower than in groups T01 (23.59, P < 0.0001) and T02 (19.37, P = 0.0001). The ability of PRRS MLV vaccines to reduce the duration of viraemia and nasal shedding after intranasal challenge with a heterologous PRRSV-1-1 strain differ significantly.

Viable cell density (VCD) and cell viability (CV) are key performance indicators of cell culture processes in biopharmaceutical production of biologics and vaccines. Traditional methods for monitoring VCD and CV involve offline cell counting assays that are both labor intensive and prone to high variability, resulting in sparse sampling and uncertainty in the obtained data. Process analytical technology (PAT) approaches offer a means to address these challenges. Specifically, in situ probe-based measurements of dielectric spectroscopy (also commonly known as capacitance) can characterize VCD and CV continuously in real time throughout an entire process, enabling robust process characterization. In this work, we propose in situ dielectric spectroscopy as a PAT tool for real time analysis of live-virus vaccine (LVV) production. Dielectric spectroscopy was collected across 25 discreet frequencies, offering a thorough evaluation of the proposed technology. Correlation of this PAT methodology to traditional offline cell counting assays was performed, in which VCD and CV were both successfully predicted using dielectric spectroscopy. Both univariate and multivariate data analysis approaches were evaluated for their potential to establish correlation between the in situ dielectric spectroscopy and offline measurements. Univariate analysis strategies are presented for optimal single frequency selection. Multivariate analysis, in the form of partial least squares (PLS) regression, produced significantly higher correlations between dielectric spectroscopy and offline VCD and CV data, as compared to univariate analysis. Specifically, by leveraging multivariate analysis of dielectric information from all 25 spectroscopic frequencies measured, PLS models performed significantly better than univariate models. This is particularly evident during cell death, where tracking VCD and CV have historically presented the greatest challenge. The results of this work demonstrate the potential of both single and multiple frequency dielectric spectroscopy measurements for enabling robust LVV process characterization, suggesting that broader application of in situ dielectric spectroscopy as a PAT tool in LVV processes can provide significantly improved process understanding. To the best of our knowledge, this is the first report of in situ dielectric spectroscopy with multivariate analysis to successfully predict VCD and CV in real time during live virus-based vaccine production.