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

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 Stream...

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Main Authors: Tan, Yan Peng, Tau, Chuan Ling, Tan, Wen Siang, Yusoff, Khatijah, Tey, Beng Ti
Format: Article
Language:English
English
Published: Elsevier 2006
Online Access:http://psasir.upm.edu.my/id/eprint/5607/1/Purification%20of%20recombinant%20nucleocapsid%20protein%20of%20Newcastle%20disease%20virus%20from%20unclarified%20feedstock%20using%20expanded%20bed%20adsorption%20chromatography.pdf
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author Tan, Yan Peng
Tau, Chuan Ling
Tan, Wen Siang
Yusoff, Khatijah
Tey, Beng Ti
author_facet Tan, Yan Peng
Tau, Chuan Ling
Tan, Wen Siang
Yusoff, Khatijah
Tey, Beng Ti
author_sort Tan, Yan Peng
collection UPM
description 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.
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spelling upm.eprints-56072016-01-21T01:56:13Z http://psasir.upm.edu.my/id/eprint/5607/ Purification of recombinant nucleocapsid protein of Newcastle disease virus from unclarified feedstock using expanded bed adsorption chromatography Tan, Yan Peng Tau, Chuan Ling Tan, Wen Siang Yusoff, Khatijah Tey, Beng Ti 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. Elsevier 2006-03 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/5607/1/Purification%20of%20recombinant%20nucleocapsid%20protein%20of%20Newcastle%20disease%20virus%20from%20unclarified%20feedstock%20using%20expanded%20bed%20adsorption%20chromatography.pdf Tan, Yan Peng and Tau, Chuan Ling and Tan, Wen Siang and Yusoff, Khatijah and Tey, Beng Ti (2006) Purification of recombinant nucleocapsid protein of Newcastle disease virus from unclarified feedstock using expanded bed adsorption chromatography. Protein Expression and Purification, 46 (1). pp. 114-121. ISSN 1046-5928 http://dx.doi.org/10.1016/j.pep.2005.06.015 English
spellingShingle Tan, Yan Peng
Tau, Chuan Ling
Tan, Wen Siang
Yusoff, Khatijah
Tey, Beng Ti
Purification of recombinant nucleocapsid protein of Newcastle disease virus from unclarified feedstock using expanded bed adsorption chromatography
title Purification of recombinant nucleocapsid protein of Newcastle disease virus from unclarified feedstock using expanded bed adsorption chromatography
title_full Purification of recombinant nucleocapsid protein of Newcastle disease virus from unclarified feedstock using expanded bed adsorption chromatography
title_fullStr Purification of recombinant nucleocapsid protein of Newcastle disease virus from unclarified feedstock using expanded bed adsorption chromatography
title_full_unstemmed Purification of recombinant nucleocapsid protein of Newcastle disease virus from unclarified feedstock using expanded bed adsorption chromatography
title_short Purification of recombinant nucleocapsid protein of Newcastle disease virus from unclarified feedstock using expanded bed adsorption chromatography
title_sort purification of recombinant nucleocapsid protein of newcastle disease virus from unclarified feedstock using expanded bed adsorption chromatography
url http://psasir.upm.edu.my/id/eprint/5607/1/Purification%20of%20recombinant%20nucleocapsid%20protein%20of%20Newcastle%20disease%20virus%20from%20unclarified%20feedstock%20using%20expanded%20bed%20adsorption%20chromatography.pdf
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