Freeze-Drying Formulations Increased the Adenovirus and Poxvirus Vaccine Storage Times and Antigen Stabilities

Abstract

Successful vaccines induce specific immune responses and protect against various viral and bacterial infections. Noninactivated vaccines, especially viral vector vaccines such as adenovirus and poxvirus vaccines, dominate the vaccine market because their viral particles are able to replicate and proliferate in vivo and produce lasting immunity in a manner similar to natural infection. One challenge of human and livestock vaccination is vaccine stability related to the antigenicity and infectivity. Freeze-drying is the typical method to maintain virus vaccine stability, while cold chain transportation is required for temperatures about 2 °C–8 °C. The financial and technological resource requirements hinder vaccine distribution in underdeveloped areas. In this study, we developed a freeze-drying formula consisting of bovine serum albumin (BSA), l -glutamic acid (L-Glu), polyethylene glycol (PEG), and dextran (DEX) to improve the thermal stability and activity of viral vaccines, including vaccinia recombinant vaccine (rTTV-OVA) and adenovirus vaccine (Ad5-ENV). We compared a panel of five different formulations (PEG: DEX: BSA: L-GLU = 50:9:0:0(#1), 50:5:4:0(#2), 50:10:9:0(#3), 50:0:0:9(#4), and 50:1:0:8(#5), respectively) and optimized the freeze-drying formula for rTTV-OVA and Ad5-ENV. We found that the freeze-drying formulations #2 and #3 could maintain rTTV-OVA infectivity at temperatures of 4 °C and 25 °C and that rTTV-OVA immunogenicity was retained during lyophilization. However, formulations #4 and #5 maintained Ad5-ENV infectivity under the same conditions, and Ad5-ENV immunogenicity had maximum retention with freeze-drying formulation #4. In summary, we developed new freeze-drying formulations that increased virus vaccine storage times and retained immunogenicity at an ambient temperature.

Single-dose replicating poxvirus vector-based RBD vaccine drives robust humoral and T cell immune response against SARS-CoV-2 infection

“…41,[69][70][71][72][73][74][75] Both vaccines are stable over a range of temperatures, which ensures easy storage and distribution, 55,56 and VACV can also be freeze-dried and stored for extended periods at 25 C without significant impact on immunogenicity. 54 In our study, the comparative analysis of humoral and cellular immune responses between TT and MVA demonstrated that TT is far more effective at stimulating neutralizing antibody production and alloreactive T cell responses against the SARS-CoV-2 RBD (Figure 2). Antibody levels were three to four orders of magnitude lower for MVA immunized mice compared with TT and T cell responses were 2-fold lower.…”

“…We investigated the short-term temperature stability of MVA and TT vaccine vectors, which has previously been demonstrated to be stable long-term under a range of temperature conditions. [54][55][56] Both viruses were stored at À80 C, 4 C, and room temperature for 1 week and then active vaccine particles were quantified by plaque assay. Under these conditions, there was no significant drop in titer TT vectors, but replication of MVA was impaired by storage at room temperature (Figure 1G).…”

A Review on Epidemiology of Monkeypox and Prevention in Thailand

“…The stabilizing efficiency is controlled by 2 factors: 1. concentration of sugars and 2. cool temperature level that freezes germs [24]. Also, there is a method of Freeze-Drying which increases stability of the Adenovirus and Poxvirus vaccine storage times [25] [26]. Still, financial and technological resources are the constraint of vaccine stability, particularly in underdeveloped regions [25].…”

“…Also, there is a method of Freeze-Drying which increases stability of the Adenovirus and Poxvirus vaccine storage times [25] [26]. Still, financial and technological resources are the constraint of vaccine stability, particularly in underdeveloped regions [25]. In extent, public health knowledge can affect preparedness of prevention [27].…”

Recent progress in drying technologies for improving the stability and delivery efficiency of biopharmaceuticals

“…In recent decades, biopharmaceuticals, including peptides, proteins, vaccines, genes, hormones, and enzymes (Sharma et al 2021 ; O’Sullivan et al 2022a , b ) have experienced rapid growth in their production (Chen et al 2021a , b ). Biopharmaceutical drugs have accounted for 35% of all FDA-approved products over the past 16 years (Vass et al 2019 ), and it is projected that by 2025, this market will be worth $395 billion (Guo et al 2020 ).…”