Self-assembly of hexahistidine-tagged tobacco etch virus capsid protein into microfilaments that induce IgG2-specific re
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Self-assembly of hexahistidine-tagged tobacco etch virus capsid protein into microfilaments that induce IgG2-specific response against a soluble porcine reproductive and respiratory syndrome virus chimeric protein Carlos Alberto Manuel-Cabrera, Alba Adriana Vallejo-Cardona, Eduardo Padilla-Camberos, Rodolfo Hernández-Gutiérrez, Sara Elisa Herrera-Rodríguez and Abel Gutiérrez-Ortega*
Abstract Background: Assembly of recombinant capsid proteins into virus-like particles (VLPs) still represents an interesting challenge in virus-based nanotechnologies. The structure of VLPs has gained importance for the development and design of new adjuvants and antigen carriers. The potential of Tobacco etch virus capsid protein (TEV CP) as adjuvant has not been evaluated to date. Findings: Two constructs for TEV CP expression in Escherichia coli were generated: a wild-type version (TEV-CP) and a C-terminal hexahistidine (His)-tagged version (His-TEV-CP). Although both versions were expressed in the soluble fraction of E. coli lysates, only His-TEV-CP self-assembled into micrometric flexuous filamentous VLPs. In addition, the His-tag enabled high yields and facilitated purification of TEV VLPs. These TEV VLPs elicited broader IgG2specific antibody response against a novel porcine reproductive and respiratory syndrome virus (PRRSV) protein when compared to the potent IgG1 response induced by the protein alone. Conclusions: His-TEV CP was purified by immobilized metal affinity chromatography and assembled into VLPs, some of them reaching 2-μm length. TEV VLPs administered along with PRRSV chimeric protein changed the IgG2/IgG1 ratio against the chimeric protein, suggesting that TEV CP can modulate the immune response against a soluble antigen. Keywords: Tobacco etch virus, Capsid protein, Virus-like particles, Hexahistidine tag, Adjuvant, Chimeric protein
Findings The structural proteins of some viruses occasionally mimic the three-dimensional nature of an actual virus while lacking the virus genome packaged inside its capsid [1]. These structures, also called virus-like particles (VLPs), apart from bearing self-assembly properties, feature highly ordered structure and surface repetitiveness, * Correspondence: [email protected] Unidad de Biotecnología Médica y Farmacéutica, Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco A.C., Normalistas 800, Colinas de la Normal, 44270 Guadalajara, Jalisco, Mexico
making them good candidates for the development of vaccines and epitope presenting platforms [2]. The upsurge of these applications has driven the cloning, expression and purification of virus structural components in a wide range of host systems (reviewed by Zeltins [3]). However, in most cases the self-assembly of viral capsid proteins (CPs) into VLPs still remains a challenge [4]. We previously attempted to explore the potential of Tobacco etch virus (TEV) particles as an adjuvant and our findings suggested that TEV induce both humoral
© The Author(s). 2016 Open Access This
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