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Shayestehfard, Kiana, Ramezani, Amin, Ghasemi, Younes, Morowvat, Mohammad Hossein. (1405). Vector Design, Cloning, and Expression of a Recombinant Immunotoxin Against Vascular Endothelial Growth Factor Containing the Recombinant Anti-Tumor Protein MAP30 in Escherichia coli. سامانه مدیریت نشریات علمی, 12(2), -. doi: 10.30476/tips.2026.112987.1367
Kiana Shayestehfard; Amin Ramezani; Younes Ghasemi; Mohammad Hossein Morowvat. "Vector Design, Cloning, and Expression of a Recombinant Immunotoxin Against Vascular Endothelial Growth Factor Containing the Recombinant Anti-Tumor Protein MAP30 in Escherichia coli". سامانه مدیریت نشریات علمی, 12, 2, 1405, -. doi: 10.30476/tips.2026.112987.1367
Shayestehfard, Kiana, Ramezani, Amin, Ghasemi, Younes, Morowvat, Mohammad Hossein. (1405). 'Vector Design, Cloning, and Expression of a Recombinant Immunotoxin Against Vascular Endothelial Growth Factor Containing the Recombinant Anti-Tumor Protein MAP30 in Escherichia coli', سامانه مدیریت نشریات علمی, 12(2), pp. -. doi: 10.30476/tips.2026.112987.1367
Shayestehfard, Kiana, Ramezani, Amin, Ghasemi, Younes, Morowvat, Mohammad Hossein. Vector Design, Cloning, and Expression of a Recombinant Immunotoxin Against Vascular Endothelial Growth Factor Containing the Recombinant Anti-Tumor Protein MAP30 in Escherichia coli. سامانه مدیریت نشریات علمی, 1405; 12(2): -. doi: 10.30476/tips.2026.112987.1367

Vector Design, Cloning, and Expression of a Recombinant Immunotoxin Against Vascular Endothelial Growth Factor Containing the Recombinant Anti-Tumor Protein MAP30 in Escherichia coli

Trends in Pharmaceutical Sciences and Technologies
دوره 12، شماره 2، شهریور 2026
نوع مقاله: Original Article
شناسه دیجیتال (DOI): 10.30476/tips.2026.112987.1367
نویسندگان
Kiana Shayestehfard1؛ Amin Ramezani2؛ Younes Ghasemi3؛ Mohammad Hossein Morowvat* 4
1Students Research Committee, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran
2Department of Medical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran
3Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran.
4Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, P.O. Box 71468-64685, Shiraz, Iran
چکیده
Age-related Macular Degeneration (AMD) heavily relies on anti-VEGF therapies, but frequent injections and resistance demand novel approaches. Recombinant immunotoxins present a promising alternative by directly targeting and eliminating pathologically active cells. This study aims to design, clone, and evaluate the expression of a novel anti-VEGF-MAP30 immunotoxin in Escherichia coli. The sequence of Ranibizumab (DrugBank: DB01270) was fused to the MAP30 sequence. Following codon optimization, the 1373 bp construct was synthesized and cloned into the pET-28a(+) expression vector. The recombinant plasmid was transformed into five E. coli strains (BL21(DE3), DH5α, SHuffle T7, XL1-Blue, and Rosetta-gami). Expression was induced using 1 mM IPTG and evaluated via SDS-PAGE across different time intervals (1 to 4 hours post-induction). Sequencing and gel electrophoresis confirmed the successful cloning of the gene. Among the evaluated hosts, E. coli BL21(DE3) exhibited the highest capacity for recombinant protein production, presenting a distinct band at 131 kDa on SDS-PAGE, whereas other strains demonstrated lower expression levels. The successful design and optimal expression of the anti-VEGF-MAP30 immunotoxin in E. coli BL21(DE3) provides a foundational step toward developing targeted therapies for ocular angiogenesis and solid tumors. Future purification and in vitro assays are warranted to validate its biological efficacy.
کلیدواژه‌ها
Cancer؛ Escherichia coli؛ Immunotoxin؛ MAP30 toxin؛ VEGF
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