کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
4752762 | 1416369 | 2017 | 29 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Construction, expression and characterization of a fusion protein HBscFv-IFNγ in Komagatella (Pichia) pastoris X33
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کلمات کلیدی
YPDSBMMYBMGYphosphate buffered saline with tween-20PVDFIFNγHBsAgperiodic acid-Schiff stainingTMBHRPYPdPBSTIACAlcohol oxidase 1AOX13,3′,5,5′-tetramethylbenzidine - 3،3 '، 5،5'-تترامیلیل بنزیدینBSA - BSANBT/BCIP - NBT / BCIPbovine serum albumin - آلبومین سرم گاوHepatitis B surface antigen - آنتی ژن سطحی هپاتیت BSDS-PAGE - الکتروفورز ژل پلی آکریل آمیدBiomedical - بیومدیکالELISA - تست الیزاEnzyme-linked immunosorbent assay - تست الیزاRoom temperature - دمای اتاقpolyvinylidene difluoride - دی فلوئورید پلی وینیلیدینYeast extract peptone dextrose medium - عصاره گیاه پپتون دی کرتروزPAS - نهHBV - هپاتیت بImmunoassay - واکنش های ایمونواسیhepatitis B virus - ویروس هپاتیت بیMonoclonal antibodies - پادتنهای تَکتیرهHorseradish peroxidase - پراکسیداز هوررادیشProtein - پروتئین
موضوعات مرتبط
مهندسی و علوم پایه
مهندسی شیمی
بیو مهندسی (مهندسی زیستی)
پیش نمایش صفحه اول مقاله
![عکس صفحه اول مقاله: Construction, expression and characterization of a fusion protein HBscFv-IFNγ in Komagatella (Pichia) pastoris X33 Construction, expression and characterization of a fusion protein HBscFv-IFNγ in Komagatella (Pichia) pastoris X33](/preview/png/4752762.png)
چکیده انگلیسی
HBscFv-IFNγ, a fusion protein constructed by fusing γ-interferon (IFNγ) with an antibody fragment HBscFv for the purpose of targeted delivery of the cytokine IFNγ, was designed in order to enhance its therapeutic efficacy through increasing its hepatoma localization. HBscFv and IFNγ were connected into HBscFv-IFNγ by the linker (Gly4Ser)3, and then the multicopy recombinant plasmids pPICZαA/(HBscFv-IFNγ)1,2,4 were constructed and transformed into Komagatella (Pichia) pastoris X33. The engineering strain X4, which had much higher copy number and could secretively express HBscFv-IFNγ, was screened from transformed X33 by qPCR. Results from SDS-PAGE, Western blotting and ELISA indicated that HBscFv-IFNγ displayed an excellent immunoreaction against HBsAg. The culture supernatant of X4 was purified by 14F7 affinity chromatography to obtain the fusion protein HBscFv-IFNγ in a purity of 95-98%. The HBscFv-IFNγ was able to bind 27.9% HBsAg in the serum of HBV transgenic mice, showing that the antibody of HBscFv-IFNγ has high binding affinity against HBsAg. The expressing of the recombinant HBscFv-IFNγ in P. pastoris provides a promising and inexpensive diagnostic reagent for preventing HBV infection.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Enzyme and Microbial Technology - Volume 102, July 2017, Pages 74-81
Journal: Enzyme and Microbial Technology - Volume 102, July 2017, Pages 74-81
نویسندگان
Ming-Hua Liang, Shi-Shui Zhou, Jian-Guo Jiang,