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کاربرد نوع شرط:
- جایگاه : پژوهشی
- مجله: Journal of Applied Biotechnology Reports
- نوع مقاله: Journal Article
- کلمات کلیدی: phage display,M13 Bacteriophage,Phage Concentration,Phage Purification,Phage Precipitation
- چکیده:
- چکیده انگلیسی: Bacteriophages, viruses which infect the bacteria are the most abundant organisms on the earth. Among them, the best studied and the most-exploited group is the filamentous phages especially M13 phage. They have shown a lot of interesting applications because of their unique features. Therefore, to get maximum performance of bacteriophage like M13, techniques need to be engaged for proper concentrations. In this review, most of these methods were explored in PubMed, Scopus and Google Scholar, using keywords including M13 bacteriophage, phage concentration, phage purification, phage display. Accordingly, the most important research papers about this subject have been collected, categorized and discussed. As a conclusion, to select an appropriate method for the concentration of M13 bacteriophages different criteria should be considered, including cost, equipment, yield and purity of the product. In general, subsequent applications of M13 phage is the most important factor for the selection of the concentration method.
- انتشار مقاله: 31-04-1398
- نویسندگان: Faezeh Fouladvand,Peyman Bemani,Mozafar Mohammadi,Razieh Amini,Farid Azizi Jalilian
- مشاهده
- جایگاه : پژوهشی
- مجله: Journal of Applied Biotechnology Reports
- نوع مقاله: Journal Article
- کلمات کلیدی: Molecular Dynamics Simulation,Molecular modeling,Acid Phosphatase,Phage ELISA,M13 pIX,SapM
- چکیده:
- چکیده انگلیسی: Phage ELISA is a common method used to confirm binding of obtained phages from phage display technique to related antigens. Enzyme-conjugated antibody directed against the major capsid protein (pVIII) or enzyme-conjugated secondary antibody against the primary antibody is used as a detection system in phage ELISA. We suggested expression of the secreted acid phosphatase (SapM) enzyme on M13 pIX minor coat protein directly, and evaluated this hypothesis using In Silico techniques. 3D structure model of the fusion protein (SapM+M13 pIX) was generated and evaluated by related software. MD simulation and TMHMM program results showed a stable fusion protein which is anchored to the inner membrane of E. col by membrane spanning region suggesting aproper assembling on M13 phage. In theory, SapM enzyme on the phage surface can catalyze the p-nitrophenyl phosphate as substrate and creates yellow color which can be measured at OD=405 nm by microtiter plate reader. We believe that decreasing the antibody layers in phage ELISA will significantly increase the reliability and reproducibility of the test and reduce its time.
- انتشار مقاله: 25-12-1394
- نویسندگان: Mozafar Mohammadi,Peyman Bemani,Neda Zarei
- مشاهده
- جایگاه : پژوهشی
- مجله: Asian Pacific Journal of Cancer Prevention
- نوع مقاله: Journal Article
- کلمات کلیدی: Apoptosis,Cancer therapy,immunoconjugate,scFv,EndoG
- چکیده:
- چکیده انگلیسی:
Aim: Immunotoxins are proteins that consist of an antibody fragment linked to a toxin, used as agents for targeted therapy of cancers. Although the most potent immunotoxins are made from bacterial and plant toxins, obstacles which contribute to poor responses are immunogenicity in patients and rapid development of neutralizing antibodies. In the present study we proposed a new therapeutic immunotoxin for targeted cancer therapy of ROR1 expressing cancers: an anti ROR1 single chain fragment variable antibody (scFv)-endonuclease G (anti ROR1 scFv-EndoG). Methods: The three-dimensional structure of anti ROR1 scFv-EndoG protein was modeled and structure validation tools were employed to confirm the accuracy and reliability of the developed model. In addition, stability and integrity of the model were assessed by molecular dynamic (MD) simulation. Results: All results suggested the protein model to be acceptable and of good quality. Conclusions: Anti-ROR1 scFv-EndoG would be expected to bind to the ROR1 extracellular domain by its scFv portion and selectively deliver non-immunogenic human endonuclease G enzyme as an end-stage apoptosis molecule into ROR1-expressing cancer cells and lead rapidly to apoptosis. We believe that anti ROR1 and other anti-tumor antigen scFv-EndoG forms may be helpful for cancer therapy.- انتشار مقاله: 14-04-1396
- نویسندگان: Peyman Bemani,Mozafar Mohammadi,Ali Hakakian
- مشاهده