کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
4753416 | 1416556 | 2017 | 8 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Lengthening of high-yield production levels of monoclonal antibody-producing Chinese hamster ovary cells by downregulation of breast cancer 1
ترجمه فارسی عنوان
طولانی شدن سطح تولید بالا تولید سلول های تخمدان هامستر چینی تولید آنتی بادی منوکلونال با کاهش سرطان سینه 1
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کلمات کلیدی
siRNA - siRNAMonoclonal antibody - آنتی بادی مونوکلونالHistone modification - اصلاح هیستونCell cycle checkpoint - ایست بازرسی چرخه سلولیChromatin remodeling - بازسازی کروماتینGene amplification - تقویت ژنRecombinant protein production - تولید پروتئین نوترکیبbreast cancer 1 - سرطان پستان 1Chinese hamster ovary cells - سلول های تخمدان هامستر چینی
موضوعات مرتبط
مهندسی و علوم پایه
مهندسی شیمی
بیو مهندسی (مهندسی زیستی)
چکیده انگلیسی
The establishment process of high-producing Chinese hamster ovary (CHO) cells for therapeutic protein production is usually laborious and time consuming because of the low probability of obtaining stable, high-producing clones over a long term. Thus, development of an efficient approach is required to establish stable, high-producing cells. This study presents a novel method that can efficiently establish sustainably high-producing cell lines by acceleration of transgene amplification and suppression of transgene silencing. The effects of breast cancer 1 (BRCA1) downregulation on gene amplification efficiency and long-term productivity were investigated in CHO cells. Small interfering RNA expression vectors against BRCA1 were transfected into the CHO DG44-derived antibody-producing cell clone. Individual cell clones were obtained after induction of gene amplification in the presence of 400Â nM methotrexate, which were cultured until passage 20. BRCA1-downregulated cell clones CHO B1Sa and B1Sb displayed 2.2- and 1.6-fold higher specific production rates than the S-Mock clone. Fluorescence in situ hybridization showed that transgene amplification occurred at a high frequency in B1Sa and B1Sb clones. Moreover, B1Sa and B1Sb clones at 20 passages had approximately 3.5- and 5.3-fold higher productivity than the S-Mock clone. Histone modification analysis revealed a decrease in an active mark for transcription, trimethylation of histone H3 at lysine 4 (H3K4), in the transgene locus of the S-Mock clone. However, H3K4 trimethylation levels were not decreased in B1Sa and B1Sb clones during long term culture. Our results suggest that high-producing cells, which maintain their productivity long-term, were efficiently established by BRCA1 downregulation.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Bioscience and Bioengineering - Volume 123, Issue 3, March 2017, Pages 382-389
Journal: Journal of Bioscience and Bioengineering - Volume 123, Issue 3, March 2017, Pages 382-389
نویسندگان
Rima Matsuyama, Noriko Yamano, Namiko Kawamura, Takeshi Omasa,