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Iranian Biomedical Journal، جلد ۲۴، شماره ۳، صفحات ۱۹۲-۲۰۰

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عنوان انگلیسی Affinity Based Nano-Magnetic Particles for Purification of Recombinant Proteins in Form of Inclusion Body
چکیده انگلیسی مقاله Background: Protein purification is the most complicated issue in the downstream processes of recombinant protein production; therefore, improved selective purification methods are important. Affinity-based protein purification method using His-tag and Ni-NTA resins is one of the most common strategies. MNPs can be used as a beneficial alternative for Ni-NTA resins. However, there is no data on the capability of MNPs for protein purification from inclusion bodies; this issue is studied here. Methods: Recombinant His-tagged proteins of EGFP-His and SK-His were expressed in E. coli BL-21 (DE3) in soluble and inclusion body forms, respectively. MNPs including Fe3O4 magnetic core, SiO2 shell, and Ni2+ on the surface were synthesized by sol-gel and hydrothermal reactions and then characterized by XRD, VSM, and SEM imaging. Both synthesized Fe3O4@NiSiO3 and Fe3O4@NixSiOy MNPs were employed to purify EGEP-His and SK-His under native and denaturing conditions, respectively. The quantity and purity of purified proteins were analyzed by micro-Bradford assay and SDS-PAGE, respectively. Results: Both synthesized MNPs were spherical and well-dispersed with the size ranging from 290 to 415 nm. Synthesized MNPs contained Fe3O4, SiO2 shell, and Ni2+ on their structures with suitable magnetization properties. Using Fe3O4@NiSiO3 and Fe3O4@NixSiOy yielded 192 and 188 µg/mg of SK-His, as compared to 207 and 195 µg/mg of EGFP-His, respectively. Conclusion: MNPs containing magnetic Fe3O4 core, SiO2 shell, and Ni2+on their surface are versatile alternatives for Ni-NTA resins in protein purification for proteins expressed in both soluble and inclusion body forms.
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نویسندگان مقاله | Masoud Seyedinkhorasani
Nano-Biotechnology Department, Pasteur Institute of Iran, Tehran, Iran


| Reza Ahangari Cohan
Nano-Biotechnology Department, Pasteur Institute of Iran, Tehran, Iran


| Saeid Taghavi Fardood
Department of Chemistry, University of Zanjan, Zanjan, Iran


| Farzin Roohvand
Virology Department, Pasteur Institute of Iran, Tehran, Iran


| Dariush Norouzian
Nano-Biotechnology Department, Pasteur Institute of Iran, Tehran, Iran


| Malihe Keramati
Nano-Biotechnology Department, Pasteur Institute of Iran, Tehran, Iran



نشانی اینترنتی http://ibj.pasteur.ac.ir/browse.php?a_code=A-10-1-806&slc_lang=other&sid=1
فایل مقاله اشکال در دسترسی به فایل - ./files/site1/rds_journals/125/article-125-2326207.pdf
کد مقاله (doi)
زبان مقاله منتشر شده other
موضوعات مقاله منتشر شده Pharmaceutical Biotechnology
نوع مقاله منتشر شده مقاله کامل
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