Thin-film freeze-drying of a bivalent Norovirus vaccine while maintaining the potency of both antigens

Development of Dry Powder Formulations of AS01Bcontaining vaccines using Thin-Film Freeze-Drying

“…Liquid AS01 B /OVA vaccine formulations were prepared using sucrose, trehalose dihydrate, 127 or D-mannitol as a stabilizer at total lipids to stabilizer ratio of 1:4, 1:8, 1:15 or 1:30 w/w , respectively. 128 First, the liquid adjuvant or vaccine formulations were frozen into thin films by single-vial TFF as 129 previously described [35, 44]. A cryogenically cooled surface was created by immersing a wide-mouth 130 glass vial (Fisher brand) in liquid nitrogen for ∼10 min.…”

“…g ., remdesivir) [35] or large molecules ( e . g ., proteins, siRNA, virus-like particles or monoclonal antibodies) [36-39] are frozen into thin films rapidly, followed be sublimation to remove the solvent [40, 41]. The TFF process achieves cooling rates ( i .…”

“…In this study, we hypothesized that thin-film freeze-drying (TFFD) technology can be used to formulate dry powders of the AS01B liposomes, alone or mixed with split virus or subunit protein antigens, while maintaining the liposome integrity and immunogenicity of the antigenic proteins upon reconstitution. In thin-film freezing (TFF), the liquid formulations of small molecules (e.g., remdesivir) [35] or large molecules (e.g., proteins, siRNA, virus-like particles or monoclonal antibodies) [36][37][38][39] are frozen into thin films rapidly, followed be sublimation to remove the solvent [40,41]. The TFF process achieves cooling rates (i.e., 100-100 K/s) intermediate between spray freeze-drying and conventional shelf freeze-drying, which are sufficient to prevent particle growth during freezing [42].…”

Mechanistic elucidation of freezing-induced surface decomposition of aluminum oxyhydroxide adjuvant

“…The use of minimum amount of protectant could reduce the complexity in vaccine formulations ( Drane et al., 2007 ; Lua et al., 2014 ). In contrast, 50% PEG 300, 10% propylene glycol, and 50% glycerol have been used as the protectants to maintain and stimulate antibody productions in hepatitis B, DTaP, or DTwP vaccines in mice ( Braun et al., 2009 ; Xue et al., 2014 ), and 5.55% sucrose has been formulated in lyophilized Norovirus vaccines as the protectant ( Xu et al., 2021 ). All together, these results indicated that octyl glucoside could effectively prevent the freezing-induced compromised immune responses.…”

“…Existing studies have suggested that the tendency of particles to aggregate could be reduced by using high concentrations of glass-forming excipients ( Clausi et al., 2008b ). Thus, trehalose and sucrose have been used as protectants in Norovirus and DTaP vaccine formulations, without causing particle aggregation or antigen loss while maintaining the relative potency of the vaccine within acceptable limits ( Xu et al., 2021 ; Xue et al., 2014 ). However, how freezing changes the intrinsic properties of the aluminum adjuvants remain elusive, which further hinders the design of effective protectants in vaccine formulations.…”