Human parvovirus B19 virus-like particles: In vitro assembly and stability

被引:45
|
作者
Sanchez-Rodriguez, Sandra P.
Muench-Anguiano, Lucia
Echeverria, Olga [2 ]
Vazquez-Nin, Gerardo [2 ]
Mora-Pale, Mauricio [3 ]
Dordick, Jonathan S. [3 ,4 ,5 ]
Bustos-Jaimes, Ismael [1 ]
机构
[1] Univ Nacl Autonoma Mexico, Lab Fis Quim & Ingn Prot, Dept Bioquim, Fac Med, Mexico City 04510, DF, Mexico
[2] Univ Nacl Autonoma Mexico, Dept Biol Celular, Fac Ciencias, Mexico City 04510, DF, Mexico
[3] Rensselaer Polytech Inst, Ctr Biotechnol & Interdisciplinary Studies, Dept Chem & Biol Engn, Troy, NY USA
[4] Rensselaer Polytech Inst, Dept Biomed Engn, Ctr Biotechnol & Interdisciplinary Studies, Troy, NY 12180 USA
[5] Rensselaer Polytech Inst, Dept Biol, Ctr Biotechnol & Interdisciplinary Studies, Troy, NY 12180 USA
关键词
Virus-like particles; Protein refolding; Nanotechnology; Virus structure; CHLOROTIC MOTTLE VIRUS; ESCHERICHIA-COLI; EMPTY CAPSIDS; VIRAL NANOPARTICLES; PROTEIN; VACCINE; GENERATION; PURIFICATION; POLYMORPHISM; EXPRESSION;
D O I
10.1016/j.biochi.2011.12.006
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Virus-like particles (VLPs) are biological nanoparticles identical to the natural virions, but without genetic material. VLPs are suitable for the analysis of viral infection mechanisms, vaccine production, tissue-specific drug delivery, and as biological nanomaterials. Human parvovirus B19 (B19) infects humans; therefore VLPs derived from this virus have enormous potential in medicine and diagnostics. Current production of self-assembled VLPs derived from B19 is typically carried out in eukaryotic expression systems. However many applications of VLPs require access to its internal core. Consequently, the processes of disassembly and further reassembly of VLPs are critical both for purification of viral proteins, and for encapsulation purposes. Herein we report the in vitro self-assembly of B19 VLPs derived from the recombinant VP2 protein expressed in Escherichia coli and the effects of pH and ionic strength on the assembly process. Our results demonstrate that VP2 is able to form VLPs completely in vitro. At neutral pH, homogeneous VLPs assemble, while at acidic and basic pHs, with low ionic strength, the major assemblies are small intermediates. The in vitro self-assembled VLPs are highly stable at 37 degrees C, and a significant fraction of particles remain assembled after 30 min at 80 degrees C. (C) 2011 Elsevier Masson SAS. All rights reserved.
引用
收藏
页码:870 / 878
页数:9
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