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

Yan Peng Tan, Tau Chuan Ling, Wen Siang Tan, Khatijah Yusoff, Beng Ti Tey

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16 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)114-121
Number of pages8
JournalProtein Expression and Purification
Volume46
Issue number1
DOIs
Publication statusPublished - Mar 2006
Externally publishedYes

Keywords

  • Dynamic binding capacity
  • Escherichia coli
  • IMA-EBAC
  • NDV
  • NP protein

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