VLP production from recombinant L1/L2 HPV-16 protein expressed in pichia pastoris

摘要: Background Human papillomavirus 16 is considered a causative agent of genital cancers. Since there is no decisive treatment, the only approach is vaccination of high-risk group. Objective This study aimed to produce a chimeric L1/L2 protein in Pichia Pastoris system. Method To develop VLPs of chimeric L1/L2 protein HPV-16, first, a cross-neutralizing epitope of HPV-16 L2 gene was inserted into L1 HPV-16 gene. Then the chimeric L1/L2 HPV-16 was inserted in pPICZA plasmid and expressed in Pichia pastoris (P. pastoris). The final purification of VLPs was carried out by ultra-centrifugation (130000 g) using 10-40% sucrose density gradient for 4 h at 4 °C. The SDS-PAGE and western blot assay was carried out for L1-HPV-16 and L2-HPV- 16 proteins separately. Amount of 55ng of the purified VLPs was coated to the wells of ELISA for detection of L1 HPV-16 antibody and L2-HPV-16 antibody by ELISA test …