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. 2018 Nov 27;9(6):e02287-18.
doi: 10.1128/mBio.02287-18.

Development of Thermostable Lyophilized Sabin Inactivated Poliovirus Vaccine

Affiliations

Development of Thermostable Lyophilized Sabin Inactivated Poliovirus Vaccine

Woo-Jin Shin et al. mBio. .

Abstract

As oral poliovirus vaccine (OPV) causes vaccine-associated paralytic poliomyelitis, the polio endgame strategy introduced by the Global Polio Eradication Initiative calls for a phased withdrawal of OPV and an introduction of inactivated poliovirus vaccine (IPV). The introduction of IPV creates challenges in maintaining the cold chain for vaccine storage and distribution. Recent advances in lyophilization have helped in finding a temperature-stable formulation for multiple vaccines; however, poliovirus vaccines have yet to capture a stable, safe formula for lyophilization. In addition, efficient in vitro methods for antigen measurement are needed for screening stable vaccine formulations. Here, we report size exclusion high-performance liquid chromatography (SE-HPLC) as a reliable means to identify the leading lyophilized formulation to generate thermostable Sabin inactivated poliovirus vaccine (sIPV). High-throughput screening and SE-HPLC determined the leading formulation, resulting in 95% D-antigen recovery and low residual moisture content of sIPV following lyophilization. Furthermore, the lyophilized sIPV remained stable after 4 weeks of incubation at ambient temperature and induced strong neutralizing antibodies and full protection of poliovirus receptor transgenic mice against the in vivo challenge of wild-type poliovirus. Overall, this report describes a novel means for the high-throughput evaluation of sIPV antigenicity and a thermostable lyophilized sIPV with in vivo vaccine potency.IMPORTANCE Poliomyelitis is a highly contagious disease caused by the poliovirus. While the live attenuated OPV has been the vaccine of choice, a major concern is its ability to revert to a form that can cause paralysis, so-called vaccine-associated paralytic poliomyelitis. Therefore, the new endgame strategy of the Global Polio Eradication Initiative includes the introduction of an IPV. However, the feasibility of the use of current IPV formulations in developing countries is limited, because IPV is insufficiently stable to be purified, transported, and stored under unrefrigerated conditions. We successfully designed the sIPV for use in the dry state that maintains the full vaccine potency in animal models after incubation at ambient temperature. This report provides, for the first time, candidate formulations of sIPV that are stable at elevated temperatures.

Keywords: D-antigen; Sabin inactivated poliovirus vaccine; cold chain; lyophilization; thermostable.

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Figures

FIG 1
FIG 1
Purified inactivated Sabin poliovirus. Polioviral particles are comprised of icosahedral capsid proteins that consist of VP1, VP2, VP3, and VP4. To check the purity of Sabin inactivated poliovirus, virions were separated using SDS-PAGE followed by silver staining. Lane 1, molecular weight standards; lane 2, before tangential flow filtration (TFF); lane 3, after TFF; lane 4, after size exclusion chromatography (SEC); lane 5, after ion-exchange chromatography (IEC).
FIG 2
FIG 2
SE-HPLC reliably measures D-antigen stability of sIPV. Poliovirus antigens are divided into D-antigen (D-Ag) and C-antigen (C-Ag); only D-Ag shows major immunogenicity. Size exclusion high-performance liquid chromatography (SE-HPLC) was used as a novel method for determining the antigenicity of sIPV through separation of intact viral particles from disintegrated capsid proteins on the basis of hydrodynamic radius. (A) sIPV chromatogram detecting 336-nm emission from 280-nm excitation through SE-HPLC. (B) SE-HPLC chromatograms of sIPV main peak eluate after 1 week of storage at 4°C in a liquid state.
FIG 3
FIG 3
Lyophilized sIPV remains stable at elevated temperatures. To test the thermostability of lyophilized sIPV, lyophilized sIPV from formulations F4, F8, and F9 was incubated at different temperatures and D-Ag recovery was measured using conventional ELISA. (A) D-Ag unit recovery over 4 weeks of incubation at 4°C. (B) D-Ag unit recovery over 4 weeks of incubation at 25°C. (C) D-Ag unit recovery over 4 weeks of incubation in 40°C.
FIG 4
FIG 4
Lyophilized sIPV effectively protects mice against wild-type poliovirus challenge. (A) Mean neutralization antibody titers of vaccination group. Blood of vaccinated cPVR transgenic mice (n = 8) treated with commercial IPOL-IPV or sIPV or reconstituted lyophilized (lyo) sIPV incubated at either 4°C or 37°C for 4 weeks was collected at day 21 to measure neutralizing antibody titers against 100 TCID50 of Sabin type 1 poliovirus. For the statistical analysis, one-way analysis of variance (ANOVA) (Kruskal-Wallis test) was used. *, P < 0.05; ***, P < 0.001. (B) In vivo vaccine efficacy of lyophilized IPV. To investigate the protective efficacy of thermostabilized sIPV in vivo, cPVR transgenic mice (n = 8) were vaccinated and boosted with commercial IPOL-IPV or sIPV or reconstituted lyophilized sIPV incubated at either 4°C or 37°C for 4 weeks. The mice were then challenged at day 28 with wild-type PV (Mahoney strain) to test virus-induced paralysis. Commercial IPOL-IPV (a trivalent polio vaccine distributed by Sanofi Pasteur) was used as a control.

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References

    1. Hogle JM, Chow M, Filman DJ. 1985. Three-dimensional structure of poliovirus at 2.9 A resolution. Science 229:1358–1365. doi:10.1126/science.2994218. - DOI - PubMed
    1. Kitamura N, Semler BL, Rothberg PG, Larsen GR, Adler CJ, Dorner AJ, Emini EA, Hanecak R, Lee JJ, van der Werf S, Anderson CW, Wimmer E. 1981. Primary structure, gene organization and polypeptide expression of poliovirus RNA. Nature 291:547–553. doi:10.1038/291547a0. - DOI - PubMed
    1. Rappuoli R, Pizza M, Del Giudice G, De Gregorio E. 2014. Vaccines, new opportunities for a new society. Proc Natl Acad Sci U S A 111:12288–12293. doi:10.1073/pnas.1402981111. - DOI - PMC - PubMed
    1. Barquet N, Domingo P. 1997. Smallpox: the triumph over the most terrible of the ministers of death. Ann Intern Med 127:635–642. doi:10.7326/0003-4819-127-8_Part_1-199710150-00010. - DOI - PubMed
    1. Plotkin S. 2014. History of vaccination. Proc Natl Acad Sci U S A 111:12283–12287. doi:10.1073/pnas.1400472111. - DOI - PMC - PubMed

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