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 & Objective:  Glioblastoma multiforme (GBM) is one of the most aggressive primary brain tumors and is characterized by a poor prognosis and limited therapeutic options. Protein tyrosine phosphatase receptor type Z1 (PTPRZ1) is overexpressed in GBM and represents a promising target for immunotherapeutic intervention.  Materials & Methods:  In the present study, Fc-fusion technology was employed to enhance the stability and potentially improve the functional activity of the recombinant protein. A PTPRZ1 fragment was inserted into the pET28a expression vector and subsequently transformed into E. coli BL21 (DE3) cells using the heat-shock method. Recombinant protein expression was optimized by evaluating different induction times (2, 4, and overnight), using an IPTG concentration of 1 mM, and incubating the cultures at 37°C for initial growth and induction. Recombinant protein expression in E. coli BL21 (DE3) was confirmed by SDS-PAGE, Bradford assay, and Western blot analysis. Results:  The SDS-PAGE analysis revealed a recombinant protein with an estimated molecular weight of about 46.3 kDa. The protein was subsequently purified under denaturing conditions using Ni-NTA affinity chromatography. The insoluble fraction exhibited the highest protein concentration, approximately 700 µg/mL, as determined by the Bradford assay at 595nm. Western blot analysis further confirmed the recombinant protein, with an approximate molecular weight of 46.3 kDa. Conclusion:  The present findings indicate that Fc-PTPRZ1 fusion proteins may serve as useful tools for the immunotherapy of glioblastoma; however, further in vivo studies are required to evaluated their immunogenicity and therapeutic potential.   PTPRZ1, Fc-fusion protein, Glioblastoma, Recombinant protein, Immunotherapy 220 229 http://journal.zums.ac.ir/browse.php?a_code=A-10-3329-4&slc_lang=en&sid=1 Zahra Hasanzadeh Shoeili Nazaninhasanzadeh1998@gmail.com 5200319475328460090730 0009-0008-7291-6444 No Zanjan Pharmaceutical Biotechnology Research Center, Zanjan University of Medical Sciences, Zanjan, Iran Neda Mohsenpour neda2363@gmail.com 5200319475328460090731 5200319475328460090731 No Zanjan Pharmaceutical Biotechnology Research Center, Zanjan University of Medical Sciences, Zanjan, Iran Taravat Talebi talebitaravat@gmail.com 5200319475328460090732 5200319475328460090732 No Zanjan Pharmaceutical Biotechnology Research Center, Zanjan University of Medical Sciences, Zanjan, Iran Ali Sharafi sharafi.a@gmail.com 5200319475328460090733 5200319475328460090733 Yes Zanjan Pharmaceutical Biotechnology Research Center, Zanjan University of Medical Sciences, Zanjan, Iran