Analytical and Preformulation Characterization Studies of Human Papillomavirus Virus-Like Particles to Enable Quadrivalent Multi-Dose Vaccine Formulation Development

Multi-Dose Formulation Development for a Quadrivalent Human Papillomavirus Virus-Like Particle-Based Vaccine: Part I - Screening of Preservative Combinations

“…Methods-Detailed descriptions of HPV VLP sample preparation are provided in detail elsewhere, 18 and a brief description is also provided in the Supplementary Methods. The experimental details of the competitive enzyme-linked immunosorbent assay (ELISA) for HPV6, 11, and 16 VLPs in solution, the microbial inhibition assay (MIC), and the modified version of the EP assay have also been described elsewhere, 18 and brief descriptions are provided in the Supplementary Methods. The details regarding the DOE experimental setup and analysis are also provided in the Supplementary Methods.…”

“…For example, Lal et al demonstrated that both 2-PE alone and a combination of MP-PP at concentrations that met the AET criteria destabilized a live attenuated rotavirus vaccine with large viral titer losses in the presence of the APs under both real-time and accelerated storage conditions. 5 Furthermore, considerations of maintaining both antigen stability and AP effectiveness (Figure 8) become much more complex with multivalent vaccine antigens (e.g., the stability profile of quadrivalent HPV vaccine containing VLP serotypes 6,11,16,18).…”

“…17 We recently described analytical characterization and preformulation studies to evaluate the effect of eight APs used in approved parenteral products (TH, 2-PE, BA, MC, PH, MP, PP, and CB) on the stability of in-solution and AH-adsorbed HPV16 VLPs. 18 Each AP led to varying levels of concentration-dependent destabilization of HPV16 VLPs under accelerated storage conditions (e.g., increased particle size, decreased mAb binding, and decreased conformational stability). As part of that work, 18 we developed two alternative antimicrobial effectiveness (AET) assays that were higher throughput and more streamlined than the timeconsuming quality control pharmacopeia assay (e.g., European pharmacopeia, Ph.…”

“…18 Each AP led to varying levels of concentration-dependent destabilization of HPV16 VLPs under accelerated storage conditions (e.g., increased particle size, decreased mAb binding, and decreased conformational stability). As part of that work, 18 we developed two alternative antimicrobial effectiveness (AET) assays that were higher throughput and more streamlined than the timeconsuming quality control pharmacopeia assay (e.g., European pharmacopeia, Ph. Eur.…”

“…Second, a statistical design-of-experiments (DOE) methodology (Response Surface Methodology Central Composite Design, RSM-CCD) was employed to predict optimal two-and three-AP combinations guided by responses from antimicrobial effectiveness and HPV16 antigen stability experiments. Based on these results described herein, the real-time and accelerated stability profiles of AH-adjuvanted quadrivalent HPV VLPs (6,11,16,18) with lead candidate AP combinations are compared to single AP-containing formulations as described in Part 2 of this work (see companion paper 17 ). The Part 1 results are discussed in terms of lessons learned for future AP screening experiments, including the pros and cons of the empirical OFAT vs. statistical DOE approaches.…”