Expression and Evaluation of HuscFv Antibody -PE40 Immunotoxin for Target Therapy of EGFR-Overexpressing Cancers


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نویسندگان: بهمن اکبری

کلمات کلیدی: Cancer target therapy, EGFR, HuscFv, Immunotoxin, Pseudomonas exotoxin A

نشریه: Iranian Journal of Biotechnology , 4 , 16 ,

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کد مقاله 13256
عنوان فارسی مقاله
عنوان لاتین مقاله Expression and Evaluation of HuscFv Antibody -PE40 Immunotoxin for Target Therapy of EGFR-Overexpressing Cancers
نوع مقاله مقاله اصیل (پژوهشی، Original)
بالاترین نمایه نامه بین‌المللی ISI
سطح مقاله هیچکدام
IF 0.338
عنوان نشریه Iranian Journal of Biotechnology
نوع نشریه داخلی ایندکس شده
شماره نشریه 4
دوره 16
تاریخ انتشار شمسی 1397/08/10
تاریخ انتشار میلادی
آدرس لینک مقاله/ همایش در شبکه اینترنت http://www.ijbiotech.com/article_80166.html
DOI
آدرس علمی (Affiliation) نویسنده متقاضی Department of Medical Biotechnology, Faculty of Medicine, Kermanshah University of Medical Sciences, Kermanshah, Iran

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Background: Epidermal growth factor receptor (EGFR) plays an important role in the progression and tumorigenesis of the various cancers. In this regards, anti-EGFR antibodies are valuable approved therapeutics for the EGFR over-expressing cancers. However, the occurrence of mutations in the EGFR and/or KRAS genes; a common phenomenon which is seen in many cancers, lead to the resistance to the EGFR-directed antibodies. EGFR based immunotoxins are capable of overcoming this limitation by directing the toxin moieties to the cancer cells resulting in cell death. Objectives: In the present study, a novel immunotoxin consisting of the truncated Pseudomonas exotoxin A (PE-40) and anti-EGFR huscFv was developed and evaluated for the induction of cell death in EGFR positive A431tumoral cells. Materials and Methods: PE-40 fragment of the exotoxin A was amplified by using PCR and ligated to pET22b-huscFv. The reaction was confirmed by PCR and restriction digestion. The immunotoxin was expressed in E. coli BL21 (plysS) and then was purified by Ni-NTA affinity column. Subsequently, the toxicity of the purified immunotoxin was evaluated on EGFR over-expressing epidermoid carcinoma of skin, A431 cell line. Results: PCR and restriction digestion experiments have verified the integrity of the immunotoxin construct. Purification by affinity column resulted in a highly purified recombinant immunotoxin. MTT assay revealed the growth inhibitory effect of the huscFv-PE40 immunotoxin on EGFR-over-expressing A431 cells with an IC50 value of 250 ng.mL-1. Conclusion: In conclusion, the results indicated that the immunotoxin developed in this study has a high toxicity on the EGFR-over-expressing tumor cells and could be considered as a promising candidate for the treatment of the EGFR positive cancers.

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نویسنده نفر چندم مقاله نویسنده مسئول
بهمن اکبریششمخير

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