Comparative Evaluation of Human Papillomavirus Type ۱۶ L۱ Protein Expressed in Plasmid- and Baculovirus-Based Systems in Insect Cells

سال انتشار: 1399
نوع سند: مقاله ژورنالی
زبان: انگلیسی
مشاهده: 95

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شناسه ملی سند علمی:

JR_ARCHRAZI-75-2_005

تاریخ نمایه سازی: 6 دی 1402

چکیده مقاله:

Human papillomavirus (HPV) has been associated with specific types of papillomas, lesions at particular anatomic sites, and malignancies. The HPV۱۶ and HPV۱۸ have been shown to play a role in a variety of carcinomas. The most documented HPV-associated cancer is cervical carcinoma. Suitable antigens are needed to be identified for the diagnostic tests and vaccines and the expression of L۱ recombinant protein should be accelerated in papillomaviruses. Therefore, in this study, the expression of the L۱ protein of HPV۱۶ was evaluated and compared in insect cells using a plasmid and a baculovirus system. The expression of the L۱ protein of HPV۱۶ in insect cells was investigated using a plasmid (InsectDirect) and a baculovirus system (BacMagic). The expressed recombinant proteins were purified from the Sf۹ lysate using Ni-NTA resin columns. The characterization of recombinant L۱ protein expressed in both systems (BacMagic and InsectDirect) was performed using immunofluorescence, SDS-PAGE, western blot, and dot blot. The yields of the purified proteins from the plasmid- and baculovirus-based systems (۱۰ ml culture; ۱۰۷ cells) had the ranges of ۴۵۵-۴۹۵ µg/ml and ۱.۴۴-۱.۶ mg/ml as analyzed by spectrophotometer, respectively. The SDS-PAGE analysis of purified proteins revealed that the recombinant proteins with the expected size of ۵۸ KDa were produced in both InsectDirect and baculovirus systems. A high degree (۹۵%) of purification was achieved using this system as observed in SDS-PAGE. The purified L۱ protein in the baculovirus system was clearly more efficient than the InsectDirect system. The results of this study indicate that the BacMagic system is an appropriate tool for large scale protein production and provides an alternative to the traditional baculovirus system. In addition, the InsectDirect system might provide a rapid and dependable pointer of whether a protein can be successfully produced in a baculovirus system. Both InsectDirect and BacMagic systems present remarkable savings in cost and time.

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نویسندگان

B. Abedi Kiasari

Iranian Veterinary Organization, Tehran, Iran

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