IMMUNOGENICITY OF MULTIEPITOPE VACCINE CANDIDATE SPIKE AND NSP3 PROTEIN FUSION SARS-COV-2 IN BALB/C MICE

• Main Author: Meliana Shani, Annisa
• Format: Final Project
• Language: Indonesia
• Subjects: Teknik (Rekayasa, enjinering dan kegiatan berkaitan)
• Online Access: https://digilib.itb.ac.id/gdl/view/78406
• Institution: Institut Teknologi Bandung

Since the outbreak of Coronavirus disease-2019 pandemic caused by Severe Acute Respiratory Syndrome-Coronavirus-2 (SARS-CoV-2) in 2019, until now World Health Organization (WHO) has not declared the end of Covid-19 pandemic. Therefore, the urgency to develop a vaccine still remain an important issue as one of the preventive strategy to eradicate the Covid- 19. The evolution of SARS-CoV-2 in the nature continues to create a possibility of the emerging new variant of SARS-CoV-2 virus which will lead to certain changes in characteristics in every variants. A mutation-resistant vaccine is needed to overcome the various possible mutation in the virus. Through the reverse vaccinology method, multiepitope vaccine can be designed which have a potential to induce both humoral and cellular immune response. In silico research has been done to design SARS-CoV-2 multiepitope vaccine using a fusion combination of the Spike and NSP3 protein. So far, the vaccine has been succesfully expressed in vitro using E.coli BL21(DE3). The aim of this research is to test the immunogencity of the vaccine candidate. The in vivo study of the vaccine candidate was carried out using mice (Mus musculus) BALB/c strain. The test conducted by injecting the combination of 60 ?g vaccine candidate with alum phosphate adjuvant in 1:1 ratio (v/v). The nontransformant E.coli BL21(DE3) protein control group of mice was injected with a combination of purified nontransformant E.coli BL21 (DE3) protein and Adju-Phos®, while the buffer control group of mice were given with PBS and Adju-Phos®, both given with the same volume and ratio regarding to the vaccine candidate test group. The vaccine candidate was administered subcutaneously at the scruff on day 0 (primary vaccination), day 14 (1st booster vaccination), and day 28 (2nd booster vaccination). Serum was collected 1 day before primary vaccination, 7 days after primary vaccination, 7 days after 1st booster vaccination, and 14 days after 2nd booster vaccination. Immunogenicity analysis of vaccine candidates was carried out using the sandwich ELISA method. The mice's body weight was monitored from one day before vaccination until 42 days after. Mice's body temperature was monitored every time before vaccination, 3 hours post-vaccination, 6 hours post-vaccination and 24 hours postvaccination. The results showed that the body temperature of mice in each group was not significantly different (P>0.05). Changes in body weight of female mice in all treatment groups were not significantly different (P>0.05). However, the male mice in the vaccine candidate group showed a significantly higher body weight compared to the nontransformant E.coli- Alum control (P=0.0007) and the PBS-Alum control (P=0.0007). The results of ELISA analysis showed that IgG antibodies were successfully formed on day 21 (P<0.05) in both male and female mice of the vaccine candidate group. Furthermore, IgG in the vaccine candidate group increased significantly on day 42 (P<0.05). Based on the research results, it can be concluded that the vaccine candidate is immunogenic in BALB/c mice.