دوشنبه 23 شهریور 1405
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دوره 34، شماره 3 - ( 4-1405 )
جلد 34 شماره 3 صفحات 229-220 |
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Ethics code: IR.ZUMS.BLC.1402.062
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Hasanzadeh Shoeili Z, Mohsenpour N, Talebi T, Sharafi A. Expression and Purification of a Recombinant Fc-PTPRZ1 Fusion Protein as a Glioblastoma Vaccine Candidate. J Adv Med Biomed Res 2026; 34 (3) :220-229
URL: http://journal.zums.ac.ir/article-1-7941-fa.html
URL: http://journal.zums.ac.ir/article-1-7941-fa.html
Expression and Purification of a Recombinant Fc-PTPRZ1 Fusion Protein as a Glioblastoma Vaccine Candidate. Journal of Advances in Medical and Biomedical Research. 1405; 34 (3) :220-229
چکیده: (432 مشاهده)
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.
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.
نوع مطالعه: مقاله پژوهشی |
موضوع مقاله:
Life Science
دریافت: 1404/12/2 | پذیرش: 1405/3/12 | انتشار: 1405/4/8
دریافت: 1404/12/2 | پذیرش: 1405/3/12 | انتشار: 1405/4/8
فهرست منابع
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