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Expression and purification of the matrix protein of Nipah virus in baculovirus insect cell system


Dezfooli, Seyedehsara Masoomi and Tan, Wen Siang and Tey, Beng Ti and Ooi, Chien Wei and Hussain, Siti Aslina (2016) Expression and purification of the matrix protein of Nipah virus in baculovirus insect cell system. Biotechnology Progress, 32 (1). pp. 171-177. ISSN 8756-7938; ESSN: 1520-6033


Nipah virus (NiV) causes fatal respiratory illness and encephalitis in humans and animals. The matrix (M) protein of NiV plays an important role in the viral assembly and budding process. Thus, an access to the NiV M protein is vital to the design of viral antigens as diagnostic reagents. In this study, recombinant DNA technology was successfully adopted in the cloning and expression of NiV M protein. A recombinant expression cassette (baculovirus expression vector) was used to encode an N-terminally His-tagged NiV M protein in insect cells. A time-course study demonstrated that the highest yield of recombinant M protein (400–500 μg) was expressed from math formula infected cells 3 days after infection. A single-step purification method based on metal ion affinity chromatography was established to purify the NiV M protein, which successfully yielded a purity level of 95.67% and a purification factor of 3.39. The Western blotting and enzyme-linked immunosorbent assay (ELISA) showed that the purified recombinant M protein (48 kDa) was antigenic and reacted strongly with the serum of a NiV infected pig.

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Additional Metadata

Item Type: Article
Subject: Nipah virus; Matrix protein; Baculovirus; Insect cells; Expression; Purification
Divisions: Faculty of Biotechnology and Biomolecular Sciences
Faculty of Engineering
Institute of Bioscience
DOI Number: https://doi.org/10.1002/btpr.2192
Publisher: American Institute of Chemical Engineers
Depositing User: Nurul Ainie Mokhtar
Date Deposited: 08 Mar 2018 04:48
Last Modified: 15 Mar 2018 01:52
Altmetrics: http://www.altmetric.com/details.php?domain=psasir.upm.edu.my&doi=10.1002/btpr.2192
URI: http://psasir.upm.edu.my/id/eprint/54246
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