IMMUNOGENICITY AND SAFETY OF A MUTIEPITOP PEPTIDE-BASED VACCINE CANDIDATE BASED ON SARS-COV-2 SPIKE AND NSP3 FUSION IN BALB/C MICE

• Main Author: Rofiqoh, Afifatur
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/85610
• Institution: Institut Teknologi Bandung

The emergence of SARS-CoV-2 since December 2019 in Wuhan, China has caused problems in various fields in countries throughout the world. Various preventive and curative efforts have been carried out to overcome health problems caused by SARS-CoV-2. Among various efforts, vaccination is the most effective effort ever made to overcome and inhibit the spread of the virus globally. Meanwhile, SARSCoV-2 has experienced many mutations and has caused several previously developed vaccines to be ineffective. One type of vaccine that is currently being developed is a peptide-based subunit vaccine. Multi-epitope vaccines are a type of peptide-based subunit vaccine consisting of several epitopes from the target pathogen that can trigger an adaptive immune response. In previous research, a multi-epitope vaccine from the spike protein and NSP3 (MEV-CoV19) was designed. The gene encoding MEV-CoV19 has also been cloned in plasmid pET- 23a(+) and transformed into E.coli BL21 cells for expression and purification. However, this vaccine candidate has not been tested in vivo. So in this study, the immunogenicity test of the MEV-CoV19 vaccine candidate was carried out in an animal model of BALB/c strain mice. Vaccination was carried out on 3 groups of mice, each group consisted of 5 male mice and 5 female mice. The first group was vaccinated with purified MEV-CoV19 protein, the second group with Supernatant Ekstrak E.coli BL21 non transforman protein and the third group with PBS as a control. In the three groups of mice, it was combined with an adjuvant in the form of aluminum phosphate with a volume ratio of 1:1. Each treatment was administered sub-cutaneously. Primary vaccination is carried out on day 1, first booster vaccination on day 14 and second booster vaccination on day 28. Meanwhile, blood serum was taken on day 0, day 7, day 21 and day 42. Euthanization and necropsy of organs such as the spleen, kidneys, liver and heart were carried out on the day 42. The immunogenicity test was carried out using the ELISA method. The experimental results showed that on day 21 there was a significant increase in the serum concentration of MEV-CoV19-specific IgM in male and female mice vaccinated with the vaccine candidate compared with mice vaccinated with Supernatant Ekstrak E.coli BL21 and PBS, and decreased on day 42. Meanwhile, the serum concentration of MEV-CoV19 specific IgG in male and female mice increased significantly on days 21 and 42 when compared with mice vaccinated with Supernatant Ekstrak E.coli BL21 and PBS (p<0.05). The results of histological observations on the kidneys, heart and liver in the three groups of mice showed minimal amounts of inflammation and necrosis and were not significantly different between treatment groups. The observation results showed that there were changes that occurred in the spleen tissue, which was characterized by widening of the white pulp and the formation of secondary follicles in the MEV-CoV19 and Supernatant Ekstrak E.coli treatment groups when compared with PBS. So from these results, it can be concluded that the MEV-CoV19 vaccine candidate has immunogenic properties and is safe in BALB/c mice.