Journal of Advances in Medical and Biomedical Research

en Expression and Purification of a Recombinant Fc-PTPRZ1 Fusion Protein as a Glioblastoma Vaccine Candidate Life Science Life Science مقاله پژوهشی Original Research Article Background and Objectives: Glioblastoma multiform is one of the most malignant primary brain tumors, which shows poor prognosis and limited therapeutic choices. Protein tyrosine phosphatase receptor type Z1 (PTPRZ1) is overexpressed in GBM and forms a promising target of immunotherapeutic intervention. Materials and Methods: In the current study, Fc-fusion technology was used to increase the stability and possible functional activity of the recombinant protein. The PTPRZ1 fragment was inserted into the pET28a expression vector and then transformed into E. coli BL21 (DE3) using the heat-shock method. Optimization of recombinant protein expression was performed by varying induction time (2, 4, and overnight hours), IPTG concentration (1 mM), and temperature (37°C for initial growth and induction). Protein expression was performed in the E. coli BL21 (DE3) and subsequently was confirmed using SDS-PAGE, Bradford, and western blot techniques. Results: The SDS-PAGE analysis revealed a recombinant protein with an estimated molecular weight of about 46.3 kDa. The protein was then purified with denaturing conditions using Ni-NTA affinity chromatography. The insoluble fraction had the highest protein concentration, approximately 700 µg/mL, as determined by the Bradford assay at 595nm. Western blot analysis also confirmed the recombinant protein, with an approximate molecular weight of 46.3 kDa. Conclusion: The current research indicates that Fc-PTPRZ1 fusion proteins could be useful for the immunotherapy of glioblastoma, but further in vivo studies are needed to assess their immunogenicity and therapeutic potential.   PTPRZ1, Fc-fusion protein, Glioblastoma, Recombinant protein, Immunotherapy 22 22 http://journal.zums.ac.ir/browse.php?a_code=A-10-3329-4&slc_lang=en&sid=1 Zahra Hasanzadeh Shoeili Nazaninhasanzadeh1998@gmail.com 5200319475328460089009 5200319475328460089009 No Zanjan Pharmaceutical Biotechnology Research Center, Zanjan University of Medical Sciences, Zanjan, Iran Neda Mohsenpour neda2363@gmail.com 5200319475328460089010 5200319475328460089010 No Zanjan Pharmaceutical Biotechnology Research Center, Zanjan University of Medical Sciences, Zanjan, Iran Taravat Talebi talebitaravat@gmail.com 5200319475328460089011 5200319475328460089011 No Zanjan Pharmaceutical Biotechnology Research Center, Zanjan University of Medical Sciences, Zanjan, Iran Ali Sharafi sharafi.a@gmail.com 5200319475328460089012 5200319475328460089012 Yes Zanjan Pharmaceutical Biotechnology Research Center, Zanjan University of Medical Sciences, Zanjan, Iran