TY - JOUR
T1 - Human parvovirus B19 virus-like particles
T2 - In vitro assembly and stability
AU - Sánchez-Rodríguez, Sandra P.
AU - Münch-Anguiano, Lucía
AU - Echeverría, Olga
AU - Vázquez-Nin, Gerardo
AU - Mora-Pale, Mauricio
AU - Dordick, Jonathan S.
AU - Bustos-Jaimes, Ismael
N1 - Funding Information:
This work was funded by grants from Instituto de Ciencia y Tecnología del Distrito Federal (ICyTDF), México ( PIFUTP08-104 ) and Consejo Nacional de Ciencia y Tecnología (CONACyT), México ( 99857 ). IBJ received support from the Alexander von Humboldt Foundation, Germany . SPSR received a PhD fellowship from CONACyT. The authors acknowledge technical support of Laura Álvarez-Añorve, Cristina Castañeda-Patlán and Silvia Juárez-Chavero from UNAM. We are grateful to Marimar López and Raymond Dove from Rensselaer Polytechnic Institute for their valuable help in sedimentation velocity analysis and TEM analysis, respectively.
PY - 2012/3
Y1 - 2012/3
N2 - 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 °C, and a significant fraction of particles remain assembled after 30 min at 80 °C.
AB - 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 °C, and a significant fraction of particles remain assembled after 30 min at 80 °C.
KW - Nanotechnology
KW - Protein refolding
KW - Virus structure
KW - Virus-like particles
UR - https://www.scopus.com/pages/publications/84857052556
U2 - 10.1016/j.biochi.2011.12.006
DO - 10.1016/j.biochi.2011.12.006
M3 - Article
C2 - 22192916
AN - SCOPUS:84857052556
SN - 0300-9084
VL - 94
SP - 870
EP - 878
JO - Biochimie
JF - Biochimie
IS - 3
ER -