title: Purification of recombinant nucleocapsid protein of Newcastle disease virus from unclarified feedstock using expanded bed adsorption chromatography.
abstract: In the present work, a single-step purification of recombinant nucleocapsid protein (NP) of the Newcastle disease virus (NDV) directly from unclarified feedstock using an expanded bed adsorption chromatography (EBAC) was developed. Streamline 25 column (ID = 25 mm) was used as a contactor and Streamline chelating adsorbent immobilized with Ni2+ ion was used as affinity adsorbent. The dynamic binding capacity of Ni2+ -loaded Streamline chelating adsorbent for the NP protein in unclarified feedstock was found to be 2.94 mg ml(-1) adsorbent at a superficial velocity of 200 cm h(-1). The direct purification of NP protein from unclarified feedstock using expanded bed adsorption has resulted in a 31% adsorption and 9.6% recovery of NP protein. The purity of the NP protein recovered was about 70% and the volume of processing fluid was reduced by a factor of 10. The results of the present study show that the IMA-EBAC developed could be used to combine the clarification, concentration and initial purification steps into a single-step operation.
authors.affiliation: Department of Chemical and Environmental Engineering, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia.
authors.forename: Yan Peng
authors.initials: YP
authors.lastname: Tan
authors.forename: Tau Chuan
authors.initials: TC
authors.lastname: Ling
authors.forename: Wen Siang
authors.initials: WS
authors.lastname: Tan
authors.forename: Khatijah
authors.initials: K
authors.lastname: Yusoff
authors.forename: Beng Ti
authors.initials: BT
authors.lastname: Tey
mesh_terms: D000327:Adsorption; D000821:Animal Feed / Q000382:microbiology; D000818:Animals; D018533:Biomass; D004926:Escherichia coli / Q000235:genetics; D009522:Newcastle disease virus / Q000201:enzymology* / Q000302:isolation & purification; D019590:Nucleocapsid Proteins / Q000235:genetics / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000302:isolation & purification; D013816:Thermodynamics
chemical_list: D019590:Nucleocapsid Proteins; D011994:Recombinant Proteins