Production of Human Papillomavirus Type-16 L1 VLP in Pichia pastoris

• Published in: Applied biochemistry and microbiology Vol. 56; no. 1; pp. 51 - 57
• Main Authors: Sanchooli, A., Aghayipour, Kh, Naghlani, Sh. Karimi, Samiee, Z., Kiasari, B. Abedi, Makvandi, M.
• Format: Journal Article
• Language: English
• Published: Moscow Pleiades Publishing 01.01.2020; Springer Nature B.V
• Subjects: animal viruses; Antibodies; Atomic force microscopy; Biochemistry; Biomedical and Life Sciences; Capsid protein; carcinogenesis; Centrifugation; Cervical cancer; Cervix; coat proteins; Developing countries; Enzyme-linked immunosorbent assay; Etiology; Gel electrophoresis; Gene expression; genes; Human papillomavirus; Human papillomavirus type 16; Invasiveness; Komagataella pastoris; L1 gene; LDCs; Life Sciences; Medical Microbiology; methanol; Microbiology; Optimization; patients; Pichia pastoris; plasmids; polyacrylamide gel electrophoresis; Proteins; Self-assembly; Sodium lauryl sulfate; Sucrose; Sugar; uterine cervical neoplasms; Vaccination; vaccines; Virus-like particles; Viruses; Western blotting; World population; human papillomavirus (HPV); HPV-16 L1 protein; virus-like particle (VLP)
• ISSN: 0003-6838; 1608-3024
• DOI: 10.1134/S0003683820010147

Human papillomavirus-16 (HPV-16) is primary etiological agent of invasive cervical cancer development in the world population. There is no treatment for HPV infection. As a possible tool for prophylactic vaccination, the development of virus-like particles (VLPs) comprising the HPV-16 L1 capsid protein is highly preferred. The expression of HPV-16 L1 gene was carried out in Pichia pastoris. It was done as a promising vaccine candidate against HVP-16 infection for developing countries. The codon optimization for HPV-16 L1 gene was done and synthesized in pPICZA plasmid. The expression of HPV-16 L1 was evaluated in P. pastoris after induction with methanol. The purification of HPV-16 VLPs was accomplished by the ultra-centrifugation using the sucrose density gradient. The sera of 101 subjects including 16 patients positive for HPV-16 and 85 individuals negative for HPV-16 were tested by ELISA assay using anti-HPV-16 antibodies. The results of ELISA test have depicted no false positive and no false negative as compared with commercial ELISA kit. The results of HPV-16 L1 expression in P. pastoris showed a band of about 56 kDa by SDS-PAGE, and were confirmed by Western-blot assay. The formation of VLP and the self-assembly of HPV-16 L1 major capsid protein in VLP was observed by transmission electron and atomic force microscopies.