title: Purification of recombinant virus-like particles of porcine circovirus type 2 capsid protein using ion-exchange monolith chromatography.
abstract: Diseases associated with porcine circovirus type 2 (PCV2) infection are having a severe economic impact on swine-producing countries. The PCV2 capsid (Cap) protein expressed in eukaryotic systems self-assemble into virus-like particles (VLPs) which can serve as antigens for diagnostics or/and as vaccine candidates. In this work, conventional adsorbents as well as a monolithic support with large pore sizes were examined for the chromatographic purification of PCV2 Cap VLPs from clarified yeast lysate. Q Sepharose XL was used for the initial separation of VLPs from residual host nucleic acids and some host cell proteins. For the further purification of PCV2 Cap VLPs, SP Sepharose XL, Heparin Sepharose CL-6B and CIMmultus SO3 monolith were tested. VLPs were not retained on SP Sepharose XL. The purity of VLPs after chromatography on Heparin Sepharose CL-6B was only 4-7% and the recovery of VLPs was 5-7%. Using ion-exchange chromatography on the CIMmultus SO3 monolith, PCV2 Cap VLPs with the purity of about 40% were obtained. The recovery of VLPs after chromatography on the CIMmultus SO3 monolith was 15-18%. The self-assembly of purified PCV2 Cap protein into VLPs was confirmed by electron microscopy. Two-step chromatographic purification procedure of PCV2 Cap VLPs from yeast lysate was developed using Q Sepharose XL and cation-exchange CIMmultus SO3 monolith.
authors.affiliation: Institute of Biotechnology, Vilnius University, Graiciuno 8, LT-02241 Vilnius, Lithuania. Electronic address: mindaugas.zaveckas@bti.vu.lt.
authors.forename: Mindaugas
authors.initials: M
authors.lastname: Zaveckas
authors.affiliation: Institute of Biotechnology, Vilnius University, Graiciuno 8, LT-02241 Vilnius, Lithuania.
authors.forename: Simas
authors.initials: S
authors.lastname: Snipaitis
authors.affiliation: Institute of Biotechnology, Vilnius University, Graiciuno 8, LT-02241 Vilnius, Lithuania.
authors.forename: Henrikas
authors.initials: H
authors.lastname: Pesliakas
authors.affiliation: Institute of Biotechnology, Vilnius University, Graiciuno 8, LT-02241 Vilnius, Lithuania.
authors.forename: Juozas
authors.initials: J
authors.lastname: Nainys
authors.affiliation: Institute of Biotechnology, Vilnius University, Graiciuno 8, LT-02241 Vilnius, Lithuania.
authors.forename: Alma
authors.initials: A
authors.lastname: Gedvilaite
mesh_terms: D000818:Animals; D036022:Capsid Proteins / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D002852:Chromatography, Ion Exchange; D017925:Circovirus / Q000235:genetics* / Q000378:metabolism; D053758:Nanoparticles / Q000378:metabolism / Q000648:ultrastructure; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D013552:Swine / Q000821:virology*; D014170:Transformation, Genetic; D014771:Virion / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure
chemical_list: D036022:Capsid Proteins; D011994:Recombinant Proteins
keywords: Ion-exchange chromatography; Monolith; Porcine circovirus type 2; Virus-like particles