title: Purification and characterization of recombinant envelope protein GP5 of porcine reproductive and respiratory syndrome virus from E. coli.
abstract: The major envelope protein, GP5, in porcine reproductive and respiratory syndrome virus (PRRSV) plays critical roles in the assembly, invasion and immune response of PRRSV particle, and is one of the mostly studied candidates in the development of recombinant vaccines. In this research, a purification process including immobilized metal affinity chromatography (IMAC) and hydrophobic interaction chromatography (HIC) was developed to prepare recombinant envelope protein GP5 with His-tag from an E. coli strain transformed with pGEM-ORF5. The result of cell culture indicated that His-tagged GP5 protein was expressed mainly in soluble form. After cell disruption, His-tagged GP5 protein was successfully purified by Ni(2+)-chelating Sepharose Fast Flow with a yield and purity of 80.5% and 48%, respectively. Recombinant GP5 protein was further purified by HIC to achieve a purity of 95%. Moreover, the purified rGP5 is shown in monomeric form contrasting the dimeric or tetrameric form when purified by a CEX-HIC process directly from PRRSV virions as reported in a previous study.
authors.affiliation: Institute of Chemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, PR China.
authors.forename: Chong
authors.initials: C
authors.lastname: Liu
authors.forename: Chun-Jiang
authors.initials: CJ
authors.lastname: Liu
authors.forename: Xi-Gang
authors.initials: XG
authors.lastname: Yuan
authors.forename: Chenming
authors.initials: C
authors.lastname: Zhang
mesh_terms: D000818:Animals; D002846:Chromatography, Affinity / Q000379:methods*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D057927:Hydrophobic and Hydrophilic Interactions; D008670:Metals / Q000737:chemistry; D019318:Porcine Reproductive and Respiratory Syndrome / Q000821:virology*; D019316:Porcine respiratory and reproductive syndrome virus / Q000737:chemistry* / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D013552:Swine; D014614:Vaccines, Synthetic / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D008670:Metals; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic; D014759:Viral Envelope Proteins
keywords: Envelope protein GP5; Histidine-tag; Hydrophobic interaction chromatography; Immobilized metal affinity chromatography; Porcine reproductive and respiratory syndrome virus