Purification of recombinant nucleocapsid protein of Newcastle disease virus from unclarified feedstock using expanded bed adsorption chromatography

• Published in: Protein expression and purification 2006-03, Vol.46 (1), p.114-121
• Main Authors: Tan, Yan Peng, Ling, Tau Chuan, Tan, Wen Siang, Yusoff, Khatijah, Tey, Beng Ti
• Format: Article
• Language: English
• Subjects: Adsorption; Animal Feed - microbiology; Animals; Biomass; Dynamic binding capacity; Escherichia coli; Escherichia coli - genetics; IMA-EBAC; NDV; Newcastle disease virus; Newcastle disease virus - enzymology; Newcastle disease virus - isolation & purification; NP protein; Nucleocapsid Proteins - genetics; Nucleocapsid Proteins - isolation & purification; Recombinant Proteins - isolation & purification; Thermodynamics
• ISSN: 1046-5928; 1096-0279
• DOI: 10.1016/j.pep.2005.06.015

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 Ni 2+ ion was used as affinity adsorbent. The dynamic binding capacity of Ni 2+-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.