Stability studies of HIV-1 Pr55gag virus-like particles made in insect cells after storage in various formulation media

Virology Journal

Edward P Rybicki²  Anna-Lise Williamson³  Ann E Meyers¹  Alisson Lynch¹ 

[1] Department of Molecular and Cell Biology, Faculty of Science, University of Cape Town, University Ave, Rondebosch, 7701, South Africa;Institute of Infectious Diseases and Molecular Medicine, Faculty of Health Sciences, University of Cape Town, Anzio Rd, Observatory, 7925, South Africa;National Health Laboratory Service, Groote Schuur Hospital, Main Rd, Observatory, 7925, South Africa

关键词: HIV vaccine;    Thermostability;    Osmolytes;    Virus-like particles;    Pr55^gag;    Human immunodeficiency virus;   

Others  :  1153758
DOI  :  10.1186/1743-422X-9-210

 received in 2012-02-28, accepted in 2012-09-17,  发布年份 2012

【 摘 要 】

Background

HIV-1 Pr55^gag virus-like particles (VLPs) expressed by baculovirus in insect cells are considered to be a very promising HIV-1 vaccine candidate, as they have been shown to elicit broad cellular immune responses when tested in animals, particularly when used as a boost to DNA or BCG vaccines. However, it is important for the VLPs to retain their structure for them to be fully functional and effective. The medium in which the VLPs are formulated and the temperature at which they are stored are two important factors affecting their stability.

Findings

We describe the screening of 3 different readily available formulation media (sorbitol, sucrose and trehalose) for their ability to stabilise HIV-1 Pr55^gag VLPs during prolonged storage. Transmission electron microscopy (TEM) was done on VLPs stored at two different concentrations of the media at three different temperatures (4°C, –20°C and −70°C) over different time periods, and the appearance of the VLPs was compared. VLPs stored in 15% trehalose at −70°C retained their original appearance the most effectively over a period of 12 months. VLPs stored in 5% trehalose, sorbitol or sucrose were not all intact even after 1 month storage at the temperatures tested. In addition, we showed that VLPs stored under these conditions were able to be frozen and re-thawed twice before showing changes in their appearance.

Conclusions

Although the inclusion of other analytical tools are essential to validate these preliminary findings, storage in 15% trehalose at −70°C for 12 months is most effective in retaining VLP stability.

【 授权许可】

   
2012 Lynch et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]UNAIDS: AIDS Epidemic Update. 2009. http://www.unaids.og/en/dataanalysis/epidemiology/2009aidsepidemicupdate webcite.
• [2]Koff WC: Accelerating HIV, vaccine development. Nature 2010, 464:161-162.
• [3]Munier CML, Andersen CR, Kelleher AD: HIV Vaccines: Progress to Date. Drugs 2011, 71:387-414.
• [4]Doan LX, Li M, Chen C, Yao Q: Virus-like particles as HIV-1 vaccines. Rev Med Virol 2003, 15:75-88.
• [5]Yao Q, Bu Z, Vzorov A, Yang C, Compans RW: Virus-like particle and DNA-based candidate AIDS vaccines. Vaccine 2003, 21:638-643.
• [6]Deml L, Speth C, Dietrich MP, Wolf H, Wagner R: Recombinant HIV-1 Pr55gag virus-like particles: potent stimulators of innate and acquired immune responses. Mol Immunol 2005, 42:259-277.
• [7]Vicente T, Roldao A, Peixoto C, Carrondo M, Alves PM: Large-scale production and purification of VLP-based vaccines. J Invert Pathol 2011, 107:42-48.
• [8]Wagner R, Fliessbach H, Wanner G, Motz G, Niedrig M, Deby G, von Brunn A, Wolf H: Studies on processing, particle formation and immunogenicity of the HIV-1 gag gene product: a possible component of a HIV vaccine. Arch Virol 1992, 127:117-137.
• [9]Jaffray A, Shephard E, van HJ, Williamson C, Williamson AL, Rybicki EP: Human immunodeficiency virus type 1 subtype C Gag virus-like particle boost substantially improves the immune response to a subtype C gag DNA vaccine in mice. J Gen Virol 2004, 85:409-413.
• [10]Chege GK, Shephard E, Meyers AE, van Harmelen JH, Williamson C, Lynch A, Gray CM, Rybicki EP, Williamson AL: HIV-1 subtype C Pr55gag virus-like particle vaccine efficiently boosts baboons primed with a matched DNA vaccine. J Gen Virol 2008, 89:2214-2227.
• [11]Chege GK, Thomas R, Shephard E, Meyers AE, Bourn W, Williamson C, Maclean J, Gray CM, Rybicki EP, Williamson AL: A prime-boost immunization regimen using recombinant BCG and Pr55gag virus-like particle vaccines based on HIV type 1 subtype C successfully elicits Gag-specific responses in baboons. Vaccine 2009, 27:4857-4866.
• [12]Ye L, Wen W, Dong K, Pan L, Bu Z, Compans RW, Zhang H, Yang C: Immunisation with a mixture of HIV Env DNA and VLP vaccines augments induction of CD8 T cell responses. J Biomed Biotechnol 2010, 10:1-11.
• [13]Galazka A, Milstien A, Zaffran J: Thermostability of vaccines. World Health Programme for Vaccines and Immunization 1998, 1-64.
• [14]Brandau DT, Jones LS, Wiethoff CM, Rexroad J, Middaugh CR: Thermal stability of vaccines. J Pharm Sci 2003, 92:218-231.
• [15]Hochachka PN, Somero GN: Biochemical Adaptation. 2002.
• [16]Bolen DW: Effects of naturally occurring osmolytes on protein stability and solubility: issues important in protein crystallization. Methods 2004, 34:312-322.
• [17]Kissman J, Ausar SF, Foubert TR, Brock J, Switzer MH, Detzi EJ, Vedivick TS, Middaugh R: Physical stabilization of Norwalk virus-like particles. J Pharm Sci 2008, 97:4208-4218.
• [18]Peixoto C, Sousa MF, Silva AC, Carrondo MJ, Alves PM: Downstream processing of triple layered rotavirus like particles. J Biotechnol 2007, 127:452-461.
• [19]Amorij J-P, Meulenaar J, Hinrichs WLJ, Stegmann T, Huckriede A, Coenen F, Frijlink HW: Rational design of an influenza subunit vaccine powder with sugar glass technology: Preventing conformational changes of haemagglutinin during freezing and freeze-drying. Vaccine 2007, 25:6447-6457.
• [20]Burgers WA, van Harmelen JH, Shephard E, Adams C, Mgwebi T, Bourn W, Hanke T, Williamson AL, Williamson C: Design and preclinical evaluation of a multigene human immunodeficiency virus type 1 subtype C DNA vaccine for clinical trial. J Gen Virol 2006, 87:399-410.
• [21]van Harmelen JH, Shephard E, Thomas R, Hanke T, Williamson AL, Williamson C: Construction and characterisation of a candidate HIV-1 subtype C DNA vaccine for South Africa. Vaccine 2003, 21:4380-4389.
• [22]Morikawa Y, Shibuya M, Goto T, Sano K: In vitro processing of human immunodeficiency virus type 1 Gag virus-like particles. Virology 2000, 272:366-374.
• [23]Bozzola JJ, Russell LD: Electron Microscopy: Principles and Techniques for Biologists. 1999.
• [24]Grgacic EVL, Anderson DA: Virus-like particles: Passport to immune recognition. Methods 2006, 40:60-65.
• [25]Pease LF, Lipin DI, Tsai DH, Zachariah MR, Lua LHL, Tarlov MJ, Middelberg APJ: Quantitative characterization of virus-like particles by asymmetrical flow field flow fractionation, electrospray differential mobility analysis, and transmission electron microscopy. Biotechnol Bioeng 2009, 102:845-855.