چاپ صفحه |  صفحه نخست سامانه |  چکیده مقاله |  نویسندگان |  دانلود مقاله | دانشگاه علوم پزشکی تبریز |
| نویسنده ثبت کننده مقاله | صفر فرج نیا |
| مرحله جاری مقاله | تایید نهایی |
| دانشکده/مرکز مربوطه | مرکز تحقیقات کاربردی دارویی |
| کد مقاله | 59630 |
| عنوان فارسی مقاله | خالص ساژی و خصوصیت یابی یک آنتی بادی تک زنجیره ای انسانی محدود کننده ی سرطان بیان کننده ی EGFRvIII |
| عنوان لاتین مقاله | Isolation and characterization of a novel human scFv inhibiting EGFRvIII expressing cancers |
| ناشر | 6 |
| آیا مقاله از طرح تحقیقاتی و یا منتورشیپ استخراج شده است؟ | بلی |
| عنوان نشریه (خارج از لیست فوق) | |
| نوع مقاله | Original Article |
| نحوه ایندکس شدن مقاله | ایندکس شده سطح یک – ISI - Web of Science |
| آدرس لینک مقاله/ همایش در شبکه اینترنت | www.elsevier.com/locate/immlet |
| EGFRvIII, a mutant form of epidermal growth factor receptor is highly expressed in glioblastoma, carci-noma of the breast, ovary, and lung but not in normal cells. This tumor specific antigen has emerged as a promising candidate for antibody based therapy of several cancers. The aim of the present study wasisolation and characterization of a human single chain antibody against EGFRvIII as a promising targetfor cancer therapy. For this, a synthetic peptide corresponding to EGFRvIII protein was used for screeningthe naive human scFv phage library. Selection was performed using a novel screening strategy for enrich-ment of rare specific clones. After five rounds of screening, six positive scFv clones against EGFRvIII wereselected using monoclonal phage ELISA, among them, a clone with an amber mutation in VH CDR2 codingsequence showed higher reactivity. The mutation was corrected through site directed mutagenesis andthen scFv fragment was expressed after subcloning into the bacterial expression vector. Expression inBL21 pLysS resulted in a highly soluble scFv appeared in soluble fraction of E. coli lysate. Bioinformaticin silico analysis between scFv and EGFRvIII sequences confirmed specific binding of desired scFv to EGFRvIII in CDR regions. The specific reactivity of the purified scFv with native EGFRvIII was confirmed by cell based ELISA and western blot. In conclusion, human anti- EGFRvIII scFv isolated from a scFv phagelibrary displayed high reactivity with EGFRvIII. The scFv isolated in this study can be the groundwork for developing more effective diagnostic and therapeutic agents against EGFRvIII expressing cancers. |
| نویسنده | نفر چندم مقاله |
|---|---|
| صفر فرج نیا | دوم |
| لیلا رهبرنیا | اول |
| نام فایل | تاریخ درج فایل | اندازه فایل | دانلود |
|---|---|---|---|
| farajnia1.pdf | 1395/10/01 | 1653075 | دانلود |
