Applying the Bioinformatic Methods to Design and Evaluate the SapM-M13 pIX Fusion Protein and Its Theoretical Role in the Phage ELISA System

سال انتشار: 1395
نوع سند: مقاله ژورنالی
زبان: انگلیسی
مشاهده: 313

فایل این مقاله در 6 صفحه با فرمت PDF قابل دریافت می باشد

استخراج به نرم افزارهای پژوهشی:

لینک ثابت به این مقاله:

شناسه ملی سند علمی:

JR_JABR-3-2_005

تاریخ نمایه سازی: 16 شهریور 1399

چکیده مقاله:

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.

نویسندگان

Mozafar Mohammadi

Applied Biotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran|Recombinant antibody laboratory, Department of Immunology, Shiraz University of Medical Sciences, Shiraz, Iran

Peyman Bemani

Recombinant antibody laboratory, Department of Immunology, Shiraz University of Medical Sciences, Shiraz, Iran

Neda Zarei

Department of Biotechnology, School of Veterinary Medicine, Shiraz University, Shiraz, Iran