Thermal Stability and Epitope Integrity of a Lyophilized Ricin Toxin Subunit Vaccine

Overview

Journal Vaccine

Specialty Allergy & Immunology

Date 2018 Sep 3

PMID 30172637

Citations 9

Authors

Jennifer Westfall

Jennifer L Yates

Greta Van Slyke

Dylan Ehrbar

Thomas Measey

Richard Straube

Oreola Donini

Nicholas J Mantis

Affiliations

Soon will be listed here.

Abstract

Biodefense vaccine are destined to be stockpiled for periods of time and deployed in the event of a public health emergency. In this report, we compared the potency of liquid and lyophilized (thermostabilized) formulations of a candidate ricin toxin subunit vaccine, RiVax, adsorbed to aluminum salts adjuvant, over a 12-month period. The liquid and lyophilized formulations were stored at stressed (40 °C) and unstressed (4 °C) conditions and evaluated at 3, 6 and 12-month time points for potency in a mouse model of lethal dose ricin challenge. At the same time points, the vaccine formulations were interrogated in vitro by competition ELISA for conformational integrity using a panel of three monoclonal antibodies (mAbs), PB10, WECB2, and SyH7, directed against known immunodominant toxin-neutralizing epitopes on RiVax. We found that the liquid vaccine under stress conditions declined precipitously within the first three months, as evidenced by a reduction in in vivo potency and concomitant loss of mAb recognition in vitro. In contrast, the lyophilized RiVax vaccine retained in vivo potency and conformational integrity for up to one year at 4 °C and 40 °C. We discuss the utility of monitoring the integrity of one or more toxin-neutralizing epitopes on RiVax as a possible supplement to animal studies to assess vaccine potency.

Citing Articles

Stressed stability and protective efficacy of lead lyophilized formulations of ID93+GLA-SE tuberculosis vaccine.

Archer M, McCollum J, Press C, Dutill T, Liang H, Fedor D Heliyon. 2023; 9(6):e17325.

PMID: 37366520 PMC: 10278894. DOI: 10.1016/j.heliyon.2023.e17325.

Safety and immunogenicity of a thermostable ID93 + GLA-SE tuberculosis vaccine candidate in healthy adults.

Sagawa Z, Goman C, Frevol A, Blazevic A, Tennant J, Fisher B Nat Commun. 2023; 14(1):1138.

PMID: 36878897 PMC: 9988862. DOI: 10.1038/s41467-023-36789-2.

Serum antibody profiling identifies vaccine-induced correlates of protection against aerosolized ricin toxin in rhesus macaques.

Roy C, Ehrbar D, Van Slyke G, Doering J, Didier P, Doyle-Meyers L NPJ Vaccines. 2022; 7(1):164.

PMID: 36526642 PMC: 9755799. DOI: 10.1038/s41541-022-00582-x.

Recombinant protein subunit SARS-CoV-2 vaccines formulated with CoVaccine HT adjuvant induce broad, Th1 biased, humoral and cellular immune responses in mice.

Lai C, To A, Wong T, Lieberman M, Clements D, Senda J Vaccine X. 2021; :100126.

PMID: 34778744 PMC: 8570651. DOI: 10.1016/j.jvacx.2021.100126.

Durable Immunity to Ricin Toxin Elicited by a Thermostable, Lyophilized Subunit Vaccine.

Novak H, Doering J, Ehrbar D, Donini O, Mantis N mSphere. 2021; 6(6):e0075021.

PMID: 34730377 PMC: 8565519. DOI: 10.1128/mSphere.00750-21.

References

Argent R, Parrott A, Day P, Roberts L, Stockley P, Lord J . Ribosome-mediated folding of partially unfolded ricin A-chain. J Biol Chem. 2000; 275(13):9263-9. DOI: 10.1074/jbc.275.13.9263. View

Smallshaw J, Firan A, Fulmer J, Ruback S, Ghetie V, Vitetta E . A novel recombinant vaccine which protects mice against ricin intoxication. Vaccine. 2002; 20(27-28):3422-7. DOI: 10.1016/s0264-410x(02)00312-2. View

Smallshaw J, Ghetie V, Rizo J, Fulmer J, Trahan L, Ghetie M . Genetic engineering of an immunotoxin to eliminate pulmonary vascular leak in mice. Nat Biotechnol. 2003; 21(4):387-91. DOI: 10.1038/nbt800. View

Olson M, Carra J, Roxas-Duncan V, Wannemacher R, Smith L, Millard C . Finding a new vaccine in the ricin protein fold. Protein Eng Des Sel. 2004; 17(4):391-7. DOI: 10.1093/protein/gzh043. View

McHugh C, Tammariello R, Millard C, Carra J . Improved stability of a protein vaccine through elimination of a partially unfolded state. Protein Sci. 2004; 13(10):2736-43. PMC: 2286567. DOI: 10.1110/ps.04897904. View

Smallshaw J, Richardson J, Pincus S, Schindler J, Vitetta E . Preclinical toxicity and efficacy testing of RiVax, a recombinant protein vaccine against ricin. Vaccine. 2005; 23(39):4775-84. DOI: 10.1016/j.vaccine.2005.04.037. View

Audi J, Belson M, Patel M, Schier J, Osterloh J . Ricin poisoning: a comprehensive review. JAMA. 2005; 294(18):2342-51. DOI: 10.1001/jama.294.18.2342. View

Vitetta E, Smallshaw J, Coleman E, Jafri H, Foster C, Munford R . A pilot clinical trial of a recombinant ricin vaccine in normal humans. Proc Natl Acad Sci U S A. 2006; 103(7):2268-73. PMC: 1413738. DOI: 10.1073/pnas.0510893103. View

Peek L, Brey R, Middaugh C . A rapid, three-step process for the preformulation of a recombinant ricin toxin A-chain vaccine. J Pharm Sci. 2006; 96(1):44-60. DOI: 10.1002/jps.20675. View

10.

Carra J, Wannemacher R, Tammariello R, Lindsey C, Dinterman R, Schokman R . Improved formulation of a recombinant ricin A-chain vaccine increases its stability and effective antigenicity. Vaccine. 2007; 25(21):4149-58. DOI: 10.1016/j.vaccine.2007.03.011. View

11.

Smallshaw J, Richardson J, Vitetta E . RiVax, a recombinant ricin subunit vaccine, protects mice against ricin delivered by gavage or aerosol. Vaccine. 2007; 25(42):7459-69. PMC: 2049008. DOI: 10.1016/j.vaccine.2007.08.018. View

12.

Rutenber E, Katzin B, Ernst S, Collins E, Mlsna D, Ready M . Crystallographic refinement of ricin to 2.5 A. Proteins. 1991; 10(3):240-50. DOI: 10.1002/prot.340100308. View

13.

Coombes L, Stickings P, Tierney R, Rigsby P, Sesardic D . Development and use of a novel in vitro assay for testing of diphtheria toxoid in combination vaccines. J Immunol Methods. 2009; 350(1-2):142-9. DOI: 10.1016/j.jim.2009.09.002. View

14.

Neal L, OHara J, Brey 3rd R, Mantis N . A monoclonal immunoglobulin G antibody directed against an immunodominant linear epitope on the ricin A chain confers systemic and mucosal immunity to ricin. Infect Immun. 2009; 78(1):552-61. PMC: 2798177. DOI: 10.1128/IAI.00796-09. View

15.

Smallshaw J, Vitetta E . A lyophilized formulation of RiVax, a recombinant ricin subunit vaccine, retains immunogenicity. Vaccine. 2010; 28(12):2428-35. PMC: 2839540. DOI: 10.1016/j.vaccine.2009.12.081. View

16.

Williamson E, Duchars M, Kohberger R . Predictive models and correlates of protection for testing biodefence vaccines. Expert Rev Vaccines. 2010; 9(5):527-37. DOI: 10.1586/erv.10.22. View

17.

OHara J, Neal L, McCarthy E, Kasten-Jolly J, Brey 3rd R, Mantis N . Folding domains within the ricin toxin A subunit as targets of protective antibodies. Vaccine. 2010; 28(43):7035-46. PMC: 2954487. DOI: 10.1016/j.vaccine.2010.08.020. View

18.

Yermakova A, Mantis N . Protective immunity to ricin toxin conferred by antibodies against the toxin's binding subunit (RTB). Vaccine. 2011; 29(45):7925-35. PMC: 3191258. DOI: 10.1016/j.vaccine.2011.08.075. View

19.

Sebaugh J . Guidelines for accurate EC50/IC50 estimation. Pharm Stat. 2012; 10(2):128-34. DOI: 10.1002/pst.426. View

20.

Vance D, Mantis N . Resolution of two overlapping neutralizing B cell epitopes within a solvent exposed, immunodominant α-helix in ricin toxin's enzymatic subunit. Toxicon. 2012; 60(5):874-7. PMC: 3461947. DOI: 10.1016/j.toxicon.2012.06.014. View