Recovery of histidine-tagged nucleocapsid protein of Newcastle disease virus using immobilised metal affinity chromatography

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

doi:10.1016/j.procbio.2005.11.003

Recovery of histidine-tagged nucleocapsid protein of Newcastle disease virus using immobilised metal affinity chromatography

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

Research output: Contribution to journal › Article › Research › peer-review

15 Citations (Scopus)

Abstract

An immobilised metal affinity packed bed adsorption chromatography (IMA-PBAC) for the purification of recombinant nucleocapsid protein (NP) of Newcastle disease virus (NDV) directly from clarified feedstock was developed. The XK 16/20 (i.d. = 16 mm) was used as a packed bed column and Streamline chelating adsorbent immobilised with Ni²⁺ ion was used as IMA adsorbent. This purification method has resulted in a 59% adsorption and 5.6% recovery of NP protein. Adsorbed NP proteins were successfully recovered using a two-step elution protocol which employed elution buffer 1 containing 50 mM imidazole to eliminate contaminating proteins and elution buffer 2 containing 350 mM imidazole to recover the NP protein at pH 8 with flow velocity of 10 cm h^-1. About 70% of the adsorbed NP protein was eluted. The purity of the recovered NP protein was about 70% and the volume of processing fluid was reduced by a factor of 4. The antigenic features of purified NP proteins were confirmed by enzyme-linked immunosorbent assay (ELISA) analysis.

Original language	English
Pages (from-to)	874-881
Number of pages	8
Journal	Process Biochemistry
Volume	41
Issue number	4
DOIs	https://doi.org/10.1016/j.procbio.2005.11.003
Publication status	Published - Apr 2006
Externally published	Yes

Keywords

Affinity chromatography
Escherichia coli
IMA
NDV
NP protein
PBA

Access to Document

10.1016/j.procbio.2005.11.003

Cite this

@article{4d0f191509594503bfcd6c6e91b32d82,

title = "Recovery of histidine-tagged nucleocapsid protein of Newcastle disease virus using immobilised metal affinity chromatography",

abstract = "An immobilised metal affinity packed bed adsorption chromatography (IMA-PBAC) for the purification of recombinant nucleocapsid protein (NP) of Newcastle disease virus (NDV) directly from clarified feedstock was developed. The XK 16/20 (i.d. = 16 mm) was used as a packed bed column and Streamline chelating adsorbent immobilised with Ni2+ ion was used as IMA adsorbent. This purification method has resulted in a 59% adsorption and 5.6% recovery of NP protein. Adsorbed NP proteins were successfully recovered using a two-step elution protocol which employed elution buffer 1 containing 50 mM imidazole to eliminate contaminating proteins and elution buffer 2 containing 350 mM imidazole to recover the NP protein at pH 8 with flow velocity of 10 cm h-1. About 70% of the adsorbed NP protein was eluted. The purity of the recovered NP protein was about 70% and the volume of processing fluid was reduced by a factor of 4. The antigenic features of purified NP proteins were confirmed by enzyme-linked immunosorbent assay (ELISA) analysis.",

keywords = "Affinity chromatography, Escherichia coli, IMA, NDV, NP protein, PBA",

author = "Tan, {Yan Peng} and Ling, {Tau Chuan} and Tan, {Wen Siang} and Khatijah Yusoff and Tey, {Beng Ti}",

note = "Funding Information: This study was supported by IRPA Grants 09-02-04-0766-EA001 and 01-02-04-003 BTK/ER/006 from the Ministry of Science, Technology, and the Innovation of Malaysia. Copyright: Copyright 2008 Elsevier B.V., All rights reserved.",

year = "2006",

month = apr,

doi = "10.1016/j.procbio.2005.11.003",

language = "English",

volume = "41",

pages = "874--881",

journal = "Process Biochemistry",

issn = "1359-5113",

publisher = "Elsevier",

number = "4",

}

TY - JOUR

T1 - Recovery of histidine-tagged nucleocapsid protein of Newcastle disease virus using immobilised metal affinity chromatography

AU - Tan, Yan Peng

AU - Ling, Tau Chuan

AU - Tan, Wen Siang

AU - Yusoff, Khatijah

AU - Tey, Beng Ti

N1 - Funding Information: This study was supported by IRPA Grants 09-02-04-0766-EA001 and 01-02-04-003 BTK/ER/006 from the Ministry of Science, Technology, and the Innovation of Malaysia. Copyright: Copyright 2008 Elsevier B.V., All rights reserved.

PY - 2006/4

Y1 - 2006/4

N2 - An immobilised metal affinity packed bed adsorption chromatography (IMA-PBAC) for the purification of recombinant nucleocapsid protein (NP) of Newcastle disease virus (NDV) directly from clarified feedstock was developed. The XK 16/20 (i.d. = 16 mm) was used as a packed bed column and Streamline chelating adsorbent immobilised with Ni2+ ion was used as IMA adsorbent. This purification method has resulted in a 59% adsorption and 5.6% recovery of NP protein. Adsorbed NP proteins were successfully recovered using a two-step elution protocol which employed elution buffer 1 containing 50 mM imidazole to eliminate contaminating proteins and elution buffer 2 containing 350 mM imidazole to recover the NP protein at pH 8 with flow velocity of 10 cm h-1. About 70% of the adsorbed NP protein was eluted. The purity of the recovered NP protein was about 70% and the volume of processing fluid was reduced by a factor of 4. The antigenic features of purified NP proteins were confirmed by enzyme-linked immunosorbent assay (ELISA) analysis.

AB - An immobilised metal affinity packed bed adsorption chromatography (IMA-PBAC) for the purification of recombinant nucleocapsid protein (NP) of Newcastle disease virus (NDV) directly from clarified feedstock was developed. The XK 16/20 (i.d. = 16 mm) was used as a packed bed column and Streamline chelating adsorbent immobilised with Ni2+ ion was used as IMA adsorbent. This purification method has resulted in a 59% adsorption and 5.6% recovery of NP protein. Adsorbed NP proteins were successfully recovered using a two-step elution protocol which employed elution buffer 1 containing 50 mM imidazole to eliminate contaminating proteins and elution buffer 2 containing 350 mM imidazole to recover the NP protein at pH 8 with flow velocity of 10 cm h-1. About 70% of the adsorbed NP protein was eluted. The purity of the recovered NP protein was about 70% and the volume of processing fluid was reduced by a factor of 4. The antigenic features of purified NP proteins were confirmed by enzyme-linked immunosorbent assay (ELISA) analysis.

KW - Affinity chromatography

KW - Escherichia coli

KW - IMA

KW - NDV

KW - NP protein

KW - PBA

UR - http://www.scopus.com/inward/record.url?scp=33344472473&partnerID=8YFLogxK

U2 - 10.1016/j.procbio.2005.11.003

DO - 10.1016/j.procbio.2005.11.003

M3 - Article

AN - SCOPUS:33344472473

SN - 1359-5113

VL - 41

SP - 874

EP - 881

JO - Process Biochemistry

JF - Process Biochemistry

IS - 4

ER -

Recovery of histidine-tagged nucleocapsid protein of Newcastle disease virus using immobilised metal affinity chromatography

Abstract

Keywords

Access to Document

Other files and links

Cite this