Expression of SARS-CoV-2 Nucleocapsid (N) Recombinant Protein Using Escherichia coli System

Rizki Aulia Ansari; Uus Saepuloh; Silmi Mariya; Yuliana; Rachmitasari Noviana; Irma Herawati Suparto; Huda Shalahudin Darusman

doi:10.4308/hjb.30.3.445-450

Expression of SARS-CoV-2 Nucleocapsid (N) Recombinant Protein Using Escherichia coli System

Rizki Aulia Ansari Graduate School Program of Biotechnology, School of Post Graduated Programs, IPB University, Bogor 16680, Indonesia
Uus Saepuloh Primate Research Center, IPB University, Bogor 16151, Indonesia
Silmi Mariya Primate Research Center, IPB University, Bogor 16151, Indonesia
Yuliana Primate Research Center, IPB University, Bogor 16151, Indonesia
Rachmitasari Noviana Primate Research Center, IPB University, Bogor 16151, Indonesia
Irma Herawati Suparto Primate Research Center, IPB University, Bogor 16151, Indonesia. Department of Chemistry, IPB University, Bogor 16680, Indonesia
Huda Shalahudin Darusman Graduate School Program of Biotechnology, School of Post Graduated Programs, IPB University, Bogor 16680, Indonesia. Primate Research Center, IPB University, Bogor 16151, Indonesia. School of Veterinary Medicine and Biomedical (SKHB), IPB University, Bogor 16680, Indonesia

DOI: https://doi.org/10.4308/hjb.30.3.445-450

Abstract

One of the main antigen that can be used for serological testing is the nucleocapsid (N) which is the most abundant viral-derived protein in SARS-CoV-2 where this virus can cause COVID19 disease. The aim of this study was to develop the SARS-CoV-2 N recombinant protein using Escherichia coli expression system. A total of 1,089 nucleotides encoding 362 amino acids of SARS-CoV-2 N was cloned to pET-14b vector. The plasmid then expressed in E. coli BL21 (DE3) and induced with 1.0 mM IPTG (Isopropyl-β-d-1-thiogalactopyranoside). The cell was harvested using denaturation lysis buffer due to inclusion body formation of SARS-CoV-2 N protein. Dialysis processed and concentrated using PEG-6000 resulted 0.992 mg/ml protein yield. Analysis of SARS-CoV-2 N recombinant protein using SDS-PAGE technique showed approximately 37.0 kDa specific band target protein. Application of this SARS-CoV-2 N recombinant protein to vaccinated and non-vaccinated antibody serum samples using ELISA technique indicated the significant result of optical density mean at 0.603 and 0.135, respectively. This study revealed that the production of SARS-COV-2 N recombinant protein could be carried out in E. coli expression system under denatured conditions, therefore the methods are more effective in producing the protein as a basic material in immuno-diagnostic assay.

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Published

2023-01-19

How to Cite

AnsariR. A., SaepulohU., MariyaS., Yuliana, NovianaR., SupartoI. H., & DarusmanH. S. (2023). Expression of SARS-CoV-2 Nucleocapsid (N) Recombinant Protein Using Escherichia coli System. HAYATI Journal of Biosciences, 30(3), 445-450. https://doi.org/10.4308/hjb.30.3.445-450

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Vol. 30 No. 3 (2023): May 2023

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