Optimization of the purification of nucleocapsid protein of newcastle disease virus by ultracentrifugation method

Protein purification is a process to remove impurities contains in a complex mixture and may involves a single step or by a combination of several steps. In this study, nucleocapsid (NP) protein of Newcastle disease virus (NDV) was purified using various purification steps from recombinant Escher...

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Main Authors:	Ahmad Raus, Raha, Yusof, Faridah
Format:	Article
Language:	English English
Published:	Journal of Pure and Applied Microbiology 2014
Subjects:	TP248.13 Biotechnology
Online Access:	http://irep.iium.edu.my/39237/ http://irep.iium.edu.my/39237/ http://irep.iium.edu.my/39237/1/P_53.pdf http://irep.iium.edu.my/39237/4/Optimization_p821-825.pdf

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spelling	iium-392372018-06-19T07:16:18Z http://irep.iium.edu.my/39237/ Optimization of the purification of nucleocapsid protein of newcastle disease virus by ultracentrifugation method Ahmad Raus, Raha Yusof, Faridah TP248.13 Biotechnology Protein purification is a process to remove impurities contains in a complex mixture and may involves a single step or by a combination of several steps. In this study, nucleocapsid (NP) protein of Newcastle disease virus (NDV) was purified using various purification steps from recombinant Escherichia coli. The purification steps involved ammonium sulphate precipitation, dialysis and sucrose gradient ultracentrifugation. In this study, the optimized condition for obtaining the highest NP protein yield in the final step of purification (ultracentrifugation) was determined. Two independent parameters which are centrifugal force and running time were manipulated and investigated. The optimum processing condition was determined by employing Face Centred Composite Design (FCCD) under Response Surface Method (RSM). Based on the almost fully elliptical plot showed in 2D and 3D response graphs, the optimum condition for obtaining the highest NP protein yield is near to the centre points of the response surface (condition of 159000xg or 31,000rpm and 5 hours). Although the optimum time to purify the highest NP protein yield is five hours centrifugation, the NP protein could also be purified after three hours centrifugation albeit with slight reduction in the yield. Less centrifugation time is important as pre- and post-centrifugation procedures for sucrose gradient ultracentrifugation is time consuming and laborious. Journal of Pure and Applied Microbiology 2014-05 Article PeerReviewed application/pdf en http://irep.iium.edu.my/39237/1/P_53.pdf application/pdf en http://irep.iium.edu.my/39237/4/Optimization_p821-825.pdf Ahmad Raus, Raha and Yusof, Faridah (2014) Optimization of the purification of nucleocapsid protein of newcastle disease virus by ultracentrifugation method. Journal of Pure and Applied Microbiology, 8 (1). pp. 821-825. ISSN 0973-7510 http://www.microbiologyjournal.org/
repository_type	Digital Repository
institution_category	Local University
institution	International Islamic University Malaysia
building	IIUM Repository
collection	Online Access
language	English English
topic	TP248.13 Biotechnology
spellingShingle	TP248.13 Biotechnology Ahmad Raus, Raha Yusof, Faridah Optimization of the purification of nucleocapsid protein of newcastle disease virus by ultracentrifugation method
description	Protein purification is a process to remove impurities contains in a complex mixture and may involves a single step or by a combination of several steps. In this study, nucleocapsid (NP) protein of Newcastle disease virus (NDV) was purified using various purification steps from recombinant Escherichia coli. The purification steps involved ammonium sulphate precipitation, dialysis and sucrose gradient ultracentrifugation. In this study, the optimized condition for obtaining the highest NP protein yield in the final step of purification (ultracentrifugation) was determined. Two independent parameters which are centrifugal force and running time were manipulated and investigated. The optimum processing condition was determined by employing Face Centred Composite Design (FCCD) under Response Surface Method (RSM). Based on the almost fully elliptical plot showed in 2D and 3D response graphs, the optimum condition for obtaining the highest NP protein yield is near to the centre points of the response surface (condition of 159000xg or 31,000rpm and 5 hours). Although the optimum time to purify the highest NP protein yield is five hours centrifugation, the NP protein could also be purified after three hours centrifugation albeit with slight reduction in the yield. Less centrifugation time is important as pre- and post-centrifugation procedures for sucrose gradient ultracentrifugation is time consuming and laborious.
format	Article
author	Ahmad Raus, Raha Yusof, Faridah
author_facet	Ahmad Raus, Raha Yusof, Faridah
author_sort	Ahmad Raus, Raha
title	Optimization of the purification of nucleocapsid protein of newcastle disease virus by ultracentrifugation method
title_short	Optimization of the purification of nucleocapsid protein of newcastle disease virus by ultracentrifugation method
title_full	Optimization of the purification of nucleocapsid protein of newcastle disease virus by ultracentrifugation method
title_fullStr	Optimization of the purification of nucleocapsid protein of newcastle disease virus by ultracentrifugation method
title_full_unstemmed	Optimization of the purification of nucleocapsid protein of newcastle disease virus by ultracentrifugation method
title_sort	optimization of the purification of nucleocapsid protein of newcastle disease virus by ultracentrifugation method
publisher	Journal of Pure and Applied Microbiology
publishDate	2014
url	http://irep.iium.edu.my/39237/ http://irep.iium.edu.my/39237/ http://irep.iium.edu.my/39237/1/P_53.pdf http://irep.iium.edu.my/39237/4/Optimization_p821-825.pdf
first_indexed	2023-09-18T20:56:21Z
last_indexed	2023-09-18T20:56:21Z
_version_	1777410329886588928

Optimization of the purification of nucleocapsid protein of newcastle disease virus by ultracentrifugation method

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