In-vitro Construction of Virus-like Particles by Purified SARS-CoV-2 Structural Proteins

S. Jhansi Rani; Sattileela SaiSujitha; A. Kalyani

doi:10.71097/IJSAT.ALSDAHW-2025.104

In-vitro Construction of Virus-like Particles by Purified SARS-CoV-2 Structural Proteins

Author(s)	S. Jhansi Rani, Sattileela SaiSujitha, A. Kalyani
Country	India
Abstract	COVID-19 is an illness caused by the coronavirus known as Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). This virus was first discovered in Wuhan, China, in December 2019. It has since become a huge health crisis in the world, as it has resulted in massive suffering and death. In order to fight these killer viruses, one needs to know their construction, life cycle, in which they infect the host cells and reproduce viral particles. An effective and efficient system of studying the virus without involving its infectious genetic material is also needed to develop appropriate therapeutic drugs and vaccines. The perfect system to be used in such studies is Virus-Like Particles (VLPs). VLPs are non-infectious particles that resemble the morphology and assembly characteristics of natural viruses without having viral nucleic acids. This renders them harmless tools to investigate virus assembly, entry and immune response mechanisms, and drug and vaccine development. In this paper, we concentrated on two significant structural proteins of SARS-CoV-2, which are the Membrane (M) protein and the Nucleocapsid (N) protein. The proteins are involved in the assembly of viruses and stabilization of their structure. M and N protein genes were cloned and produced with the help of a bacterial expression system. The purified proteins were extracted using Ni-NTA affinity chromatography, which is the selective binding and purification of the protein of interest. After purification, dialysis of the proteins against the urea buffer was done to re-establish the natural structure of the proteins. Refolded M and N protein was then incubated under set conditions in-vitro to promote Virus-Like Particles assembly. The fact that M and N proteins allowed the formation of SARS-CoV-2 VLPs successfully revealed that they can self-assemble and offers a great opportunity in future studies. They can be used in examining different viral components and functions, screening antiviral agents, and aiding in the establishment of successful treatment and prevention methods against SARS-CoV-2 and other coronaviruses.
Keywords	SARS-CoV-2, Virus-like Particles (VLPs), M Protein, N Protein, Cloning, Expression, Ni-NTA Affinity Chromatography, Therapeutic Development
Published In	Conference / Special Issue (Volume 17 \| Issue 1) - One Day National Seminar on “Advances in Life Sciences for Diversity, Applications, and Human Welfare” (ALSDAHW-2025) (March 2026)
Published On	2026-03-16
DOI	https://doi.org/10.71097/IJSAT.ALSDAHW-2025.104

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In-vitro Construction of Virus-like Particles by Purified SARS-CoV-2 Structural Proteins

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