VLP production in Leishmania tarentolae : A novel expression system for purification and assembly of HPV16 L1
| Autor: | Azam Bolhassani, Elnaz Agi, Noushin Davoudi, Zeinab Shirbaghaee |
|---|---|
| Rok vydání: | 2015 |
| Předmět: |
Immunogen
viruses Genetic Vectors Gene Expression Biology complex mixtures law.invention Papillomavirus Vaccines Western blot law Gene expression medicine Animals Humans Vaccines Virus-Like Particle Leishmania Human papillomavirus 16 medicine.diagnostic_test Papillomavirus Infections virus diseases Oncogene Proteins Viral Transfection biology.organism_classification Virology Recombinant Proteins Mice Inbred C57BL Antibody Formation Humoral immunity Recombinant DNA Capsid Proteins Female Immunization Biotechnology |
| Zdroj: | Protein Expression and Purification. 116:7-11 |
| ISSN: | 1046-5928 |
| DOI: | 10.1016/j.pep.2015.08.024 |
| Popis: | Viral like particles (VLPs) have been used as immunogen for improvement of preventive vaccines against several viral infections in preclinical and clinical trials. These constructs can stimulate both cellular and humoral immunity. Two prophylactic HPV L1 VLP vaccines known as Gardasil and Cervarix were commercialized worldwide. However, there are main problems for expression and purification of VLPs in eukaryotic expression systems such as baculovirus and yeast leading to high cost of these vaccines. A novel Leishmania protozoan system has been applied to produce different recombinant proteins due to unique properties including generation of similar proteins with mammalian, easy handling, and large-scale culture. In the current study, we developed a novel strategy to produce HPV L1 VLP using stably transfected Leishmania cells. The positive transfectants were analyzed by SDS-PAGE and Western blot analysis. The assembly of purified L1 protein was detected by TEM microscopy. Finally, C57BL/6 mice were immunized by crude VLPs and antibody responses were assessed. The results of electronic microscopy revealed average 55-60 nm for L1 VLP. Furthermore, high IgG1 and IgG2a antibody responses were generated by L1 VLPs in mice similar to L1 VLPs produced in baculovirus-infected insect cells. Regarding the results, the amount of recombinant protein generated by Leishmania was 2-3mg/500 ml media, suggesting further optimization of this system for using in large animals and human. |
| Databáze: | OpenAIRE |
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