Studi Waktu Infeksi Virus-Like Particle (Vlp) Sars-Cov-2 Pada Sel 16 Hbe14o

Rohmah, Ilmiana Nurur and Prof. Dr. Ir. Moch. Sasmito, MS., IPU., ASEAN. Eng and Prof. Widodo, S.Si., M.Si., Ph.D.Med.Sc (2024) Studi Waktu Infeksi Virus-Like Particle (Vlp) Sars-Cov-2 Pada Sel 16 Hbe14o. Magister thesis, Universitas Brawijaya.

Abstract

Virus-like Particle (VLP) merupakan struktur dari turunan virus yang terdiri dari satu atau lebih molekul berbeda dengan kemampuan untuk merakit sendiri (self-assemble). VLP memiliki peluang tinggi internalisasi yang baik karena luas permukaannya yang besar dan banyaknya komponen residu asam amino di permukaannya. VLP digunakan sebagai solusi menjanjikan dalam menangani pandemi COVID-19 dengan membuat Virus-like Particle (VLP) SARS-CoV-2 sebagai strategi dalam penemuan obat melawan COVID-19. VLP SARS-CoV-2 dibentuk dan diproduksi dari plasmid pcDNA 3.1. Plasmid yang dikonstruksikan mengandung sekuen nukleutida dari struktural protein SARS-CoV-2 yaitu Spike, Membran dan Envelope. Protein Spike tersebut di konstruksi dengan marker, spike-EGFP (S-EGFP). VLP SARS-CoV-2 yang telah diproduksi dan dikarakterisasi sebelumnya, dilihat kemampuan waktu internalisasinya kedalam sel line 16 HBE14o-. Sel line 16HBE14o- memiliki kemampuan internalisasi VLP yang lebih tinggi, karena memiliki resptor potensial ACE2 yang lebih banyak dan TMPRRS dari epitel tubuh lainnya. Tujuan penelitian ini adalah mempelajari waktu dan konsentrasi optimum yang dibutuhkan internalisasi VLP SARS-CoV-2 kedalam sel inang, serta memberikan acuan konsentrasi VLP SARS-CoV-2 dan waktu inkubasi internalisasi optimum VLP SARS-CoV-2.

English Abstract

Virus-like particles (VLP) are structures from virus derivatives that consist of one or more different molecules with the ability to self-assemble. VLPs have a high chance of good internalization due to their large surface area and the large number of amino acid residue components on their surface. VLP is used as a promising solution in dealing with the COVID-19 pandemic by creating SARS-CoV-2 Virus-like Particles (VLP) as a strategy in drug discovery against COVID-19. SARS-CoV-2 VLPs are formed and produced from the pcDNA 3.1 plasmid. The constructed plasmid contains the nucleotide sequences of the SARS-CoV-2 structural proteins, namely Spike, Membrane and Envelope. The Spike protein was constructed with the marker, spike-EGFP (S-EGFP). The SARS-CoV-2 VLP that had been previously produced and characterized was examined for its ability to internalize into cell line 16 HBE14o-. The 16HBE14o- cell line has a higher VLP internalization ability, because it has more potential ACE2 receptors and TMPRRS than other body epithelium. The aim of this research is to study the optimum time and concentration required for internalization of SARS-CoV-2 VLPs into host cells, as well as providing a reference for the concentration of SARS-CoV-2 VLPs and the optimum incubation time for internalization of SARS-CoV-2 VLPs. The method used uses SARS-CoV-2 VLPs cultured and isolated from the HEK 293T cell line. Internalization of SARS-CoV-2 VLPs was carried out during the specified time points (6, 12, 24 and 48 hours) by adding SARS-CoV-2 VLP concentrations of 2.5, 25, 50, 100, 200, 400, 600 µg/µl into the medium cell culture 16 HBE14o-. Confirmation of SARS-CoV-2 VLP internalization was carried out by observing the glow of viral EGFP (S-EGFP) with a wavelength of 488 nm, using Flow cytometry and Confocal Laser Scanning Microscopy (CLSM). Internalization positive cell data was analyzed using twoway ANOVA to see the significance of the data displayed in graphical form.

Item Type:	Thesis (Magister)
Identification Number:	0424090033
Divisions:	S2/S3 > Magister Biologi, Fakultas MIPA
Depositing User:	Unnamed user with username nova
Date Deposited:	07 Oct 2024 03:35
Last Modified:	07 Oct 2024 03:35
URI:	http://repository.ub.ac.id/id/eprint/228498

Text (DALAM MASA EMBARGO) Ilmiana Nurur Rohmah.pdf Restricted to Registered users only Download (3MB)

Actions (login required)

View Item