META-ANALYSIS OF HPV L1 EXPRESSION AND CONSTRUCTION OF HPV 52 L1 EXPRESSION SYSTEM ON PICHIA PASTORIS GS115

• Main Author: Hendrawan, Andre
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/79505
• Institution: Institut Teknologi Bandung

Human papillomavirus is well-known as the risk factor of cervical cancer. However, the coverage of HPV vaccination in Indonesia remains low due to high-cost vaccination. One of the hindrances leading to high vaccination cost is the low rate of L1 production. To date, the L1 antigen has been produced using a variety of organisms with different rates and immunogenicity. Thus, this study was conducted to perform the meta-analysis of L1 gene expression and L1 immunogenicity (represented by post-injection anti-L1 IgG titre in mice (Mus musculus)) of varying organism hosts, and the construction of L1 gene expression system in Pichia pastoris GS115. Meta-analysis was performed by processing the data of L1 titre, induction period, and anti-L1 IgG titre extracted from 19 articles which have passed the articles screening. The search terms used in articles searching were "human papillomavirus" AND "L1" AND "expression". The data of L1 titre and expression induction period were used to calculate the L1 production rate for each organism. Statistical significance on the difference of L1 expression rate and IgG titre were determined through t-test. Pearson’s correlation analysis was conducted to determine the treatments affecting the L1 expression rate and IgG titre. In construction of L1 gene expression, the pPICZ A inserted by L1 gene was cloned and linearised. Then, P. pastoris cell was transformed using the linearised recombinant plasmid. The result of meta-analysis revealed weak evidence that E. coli produced L1 with the highest rate on 95% confidence level. The highest anti-L1 IgG titre was induced by the L1 expressed using Saccharomyces cerevisiae, even though the data evidence could not support this claim on 95% confidence level. Pearson’s correlation analysis showed that the concentration of glucose, IPTG, NH4+, K+, Ca2+, Mn2+, Fe2+, Zn2+, B4O72?, H2PO4?, HPO42?, Mo7O246? and citric acid had a positive correlation with L1 production rate in E. coli. In E. coli, Saccharomyces cerevisiae, and P. pastoris, injection doses had the positive correlation with IgG titre. In the construction of L1 gene expression system, electrophoregram showed that cloning and linearisation has been successfully performed. P. pastoris GS115 has been transformed using cloned and linearised recombinant plasmid. However, the success of transformation has not been confirmed yet.