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کاربرد نوع شرط:
- جایگاه : پژوهشی
- مجله: Iranian Journal of Biotechnology
- نوع مقاله: Journal Article
- کلمات کلیدی: eCG, Ouchterlony,Polyoclonal Antibody,Solid-Phase Extraction (SPE)
- چکیده:
- چکیده انگلیسی: Background: Equine chorionic gonadotropin (eCG) is commonly used in association with a progestagen treatment to synchronize estrus of goats and ewes during breeding and non-breeding seasons. Classical purification of the eCG from serum includes pH fractionation with metaphosphoric acid two ethanol precipitation steps as well as dialysis followed by fixed-bed chromatography. Objectives: The aim of the current study was to develop an accurate and fast method for production and purification of eCG using polyoclonal antibody assay. Materials and Methods: The blood samples (300 ml) were taken from jugular vein of 17 mares on days 50, 70 and 90 of pregnancy. Plasma of the samples was siphoned and phenol solution has been added to the plasma and stored in the refrigerator until eCG extraction. To prepare polyoclonal antibody against eCG, four male rabbits, about 4-months-old and 2-kg weights, were chosen. A basic immunization was done by injecting 25 IU of eCG to the rabbits. Ouchterlony assay or double immunodiffusion test was used to assess the immunization and the titer of antiserum against eCG. eCG has been purified from the plasma via Solid-Phase Extraction (SPE) method. Results: Results based on agarose-gel double immunodiffusion test showed that rabbits have been completely immunized. SDS-PAGE analysis showed purified eCG is extracted without any significant contamination. Conclusions: The extraction of eCG with polyclonal antibody using SPE method and production of anti- eCG antiserum in rabbits is suitable and may be a cost effective method for large scale production of eCG and anti- eCG antiserum in Iran.
- انتشار مقاله: 09-07-1392
- نویسندگان: Mohammad Ali Sharif,Hamid Kohram,Ahmad Zare Shahneh,Hossein Zolfagharian,Bahman Abedi Kiasari,Mehdi Hedayati
- مشاهده
- جایگاه : پژوهشی
- مجله: Archives of Razi Institute
- نوع مقاله: Journal Article
- کلمات کلیدی: Cloning,Expression,Baculovirus,Papillomavirus
- چکیده:
- چکیده انگلیسی: Human papillomavirus (HPV) has been associated with specific types of papillomas, lesions at particular anatomic sites, and malignancies. The HPV16 and HPV18 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 L1 recombinant protein should be accelerated in papillomaviruses. Therefore, in this study, the expression of the L1 protein of HPV16 was evaluated and compared in insect cells using a plasmid and a baculovirus system. The expression of the L1 protein of HPV16 in insect cells was investigated using a plasmid (InsectDirect) and a baculovirus system (BacMagic). The expressed recombinant proteins were purified from the Sf9 lysate using Ni-NTA resin columns. The characterization of recombinant L1 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 (10 ml culture; 107 cells) had the ranges of 455-495 µg/ml and 1.44-1.6 mg/ml as analyzed by spectrophotometer, respectively. The SDS-PAGE analysis of purified proteins revealed that the recombinant proteins with the expected size of 58 KDa were produced in both InsectDirect and baculovirus systems. A high degree (95%) of purification was achieved using this system as observed in SDS-PAGE. The purified L1 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.
- انتشار مقاله: 23-05-1397
- نویسندگان: B. Abedi Kiasari,B. Abedi Kiasari,B. Abedi Kiasari
- مشاهده
- جایگاه : پژوهشی
- مجله: Archives of Razi Institute
- نوع مقاله: Journal Article
- کلمات کلیدی: Epidemiology,Polyomavirus,JCV,PML,HAART
- چکیده:
- چکیده انگلیسی: Polyomaviruses may cause human disease, particularly in immunocompromised hosts. JCV, one of the members of polyomaviridae family, is the causative agent of the neurological disease progressive multifocal leukoencephalopathy (PML), which occurs mostly in immunocompromised patients. Progressive Multifocal leukoencephalopathy (PML) is a progressive demyelinating disorder of the central nervous system (CNS) resulting from infection of oligodendrocytes by JC polyomavirus. Whereas after highly active antiretroviral therapy (HAART), the incidence of nearly all of all neurological complication have decreased, this article evaluates the incidence and prognosis of JC polyomavirus-associated PML including a unchangeable incidence of PML associated with JC polyomavirus compared with other opportunistic disease and paradoxical results of disease remission in HAART era.
- انتشار مقاله: 27-09-1393
- نویسندگان: B Abedi Kiasari,B Abedi Kiasari,B Abedi Kiasari
- مشاهده