Bac to Bac System Efficiency for Preparing HPV Type ۱۶ Virus-Like Particle Vaccine
محل انتشار: مجله آرشیو رازی، دوره: 78، شماره: 3
سال انتشار: 1402
نوع سند: مقاله ژورنالی
زبان: انگلیسی
مشاهده: 55
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شناسه ملی سند علمی:
JR_ARCHRAZI-78-3_025
تاریخ نمایه سازی: 6 دی 1402
چکیده مقاله:
Today, the human papillomavirus (HPV) L۱ protein is the main target in the construction of prophylactic HPV vaccines. The production of virus-like particles (VLPs) that closely resemble the natural structure of the HPV۱۶ virus and induce high levels of virus-neutralizing antibodies in animals and humans is facilitated by the expression of HPV۱۶-L۱ protein in eukaryotic cells. The Bac-to-Bac system has been previously used to produce high levels of recombinant proteins. In this study, we utilized this expression system to generate HPV۱۶-L۱ VLPs in Spodoptra frugipedra (Sf۹) insect cells. The wild-type L۱ gene of papillomavirus type ۱۶ was selected from Gene Bank and placed in bacmid structure after codon optimization using pFast Bac vector. The recombinant baculovirus containing HPV-۱۶/L۱ gene was then provided using the Bac-to-Bac system. It should be mentioned that the vector was transfected into the Sf۹ cell. The cells were then lysed and the expression of L۱ protein was revealed by SDS-PAGE and confirmed by Western Blot. The L۱ purification was performed through Ni-NTA chromatography. The VLP formation of papillomavirus L۱ protein was visualized by transmission electron microscopy. The expressed recombinant L۱ was ~۶۰ KD on SDS-PAGE which was identified in western blot by a specific anti-L۱ monoclonal antibody. The electron microscopy confirmed the assembly of VLPs. Results of this study showed that the production of this protein at the industrial level can be optimized using a baculovirus/Sf۹ system. The characteristics and advantages of this system are promising and it is a suitable candidate for protein synthesis.
کلیدواژه ها:
نویسندگان
H Razavi-Nikoo
Department of Microbiology, Golestan University of Medical Sciences, Gorgan, Iran
E Behboudi
Department of Basic Sciences, Khoy University of Medical Sciences, Khoy, Iran
B Aghcheli
Department of Microbiology, Golestan University of Medical Sciences, Gorgan, Iran
S. M. A Hashemi
Department of Microbiology, Golestan University of Medical Sciences, Gorgan, Iran
A Moradi
Department of Microbiology, Golestan University of Medical Sciences, Gorgan, Iran
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