Ebook Update on production of recombinant therapeutic protein – Transient gene expression: Part 2
➤ Gửi thông báo lỗi ⚠️ Báo cáo tài liệu vi phạmNội dung chi tiết: Ebook Update on production of recombinant therapeutic protein – Transient gene expression: Part 2
Ebook Update on production of recombinant therapeutic protein – Transient gene expression: Part 2
3 Optimisation of Transient Gene Expression for Therapeutic Protein ProductionJianwei ZhuTransient gene expression (TGE) is a well-established technol Ebook Update on production of recombinant therapeutic protein – Transient gene expression: Part 2logy for rapid generation of recombinant proteins with human embryonic kidney (HEK) and Chinese hamster ovary (CHO) cell lines. Many TGE protocols have been published over the last ten years. A number of representative protocols are described in the last section of Chapter 2 for producing therapeuti Ebook Update on production of recombinant therapeutic protein – Transient gene expression: Part 2c proteins at different scales (from 10 mL -100 L), facilitated by calcium phosphate or polyethylenimine (PEI), using viral or non-viral vectors. TheEbook Update on production of recombinant therapeutic protein – Transient gene expression: Part 2
protocol using 25 kDa linear PEI (Table 2.3) as a transfection component to form a plasmid/ PEI complex for transfecting CHO and HEK cells has been wi3 Optimisation of Transient Gene Expression for Therapeutic Protein ProductionJianwei ZhuTransient gene expression (TGE) is a well-established technol Ebook Update on production of recombinant therapeutic protein – Transient gene expression: Part 2t few years to the extent that it is now feasible to produce 100 g of protein for preclinical studies or even early phase clinical development of therapeutics. Successful reports from the past few years are listed and summarised in Table 3.1.Table 3.1 presents the representative productivity level w Ebook Update on production of recombinant therapeutic protein – Transient gene expression: Part 2ith the TGE technology platform. Since 2008, when the production titre first reached the 1 g/L milestone 11 J, there have been repeatedly successful rEbook Update on production of recombinant therapeutic protein – Transient gene expression: Part 2
esults either through TGE [21 or stable transfection pools [3] at, or over, 1 g/L expression levels, making it feasible to use the TGE system to produ3 Optimisation of Transient Gene Expression for Therapeutic Protein ProductionJianwei ZhuTransient gene expression (TGE) is a well-established technol Ebook Update on production of recombinant therapeutic protein – Transient gene expression: Part 2h CHO and HEK293 cells were predominantly used as host cell lines. Great effort has been invested into optimisation of many aspects of the TGE system in coexpression of antiapoptosis genes [1], the use of new additives in the media [4,5], and transfection81coTable 3.1 Recombinant therapeutics produc Ebook Update on production of recombinant therapeutic protein – Transient gene expression: Part 2ed by transient gene expression technology platformCellExpressionProductExpression and process descriptionReferenceHEK293E81 mg/LMabEnhancement by cotEbook Update on production of recombinant therapeutic protein – Transient gene expression: Part 2
ransfection of WPRE by five-fold1531HEK293E1000 mg/LMab1) Human CMV + Intron + WPRE (40 mg/L)1112) Cotransfection with 37.5% HC, 10% LC, 10% hpl810% h3 Optimisation of Transient Gene Expression for Therapeutic Protein ProductionJianwei ZhuTransient gene expression (TGE) is a well-established technol Ebook Update on production of recombinant therapeutic protein – Transient gene expression: Part 2ed in IIEK293E cells compared with pcDNA3.1 vector.1751CHOK1SV80 mg/LMab1) DMSO and lithium acetate increased expression1512) Fed-batch process maintained for 14 days with 3 post-transfections at 2-4 day intervals3) Product showed similar glycosylation patterns from batch to batch4) Process scaled u Ebook Update on production of recombinant therapeutic protein – Transient gene expression: Part 2p to 20 L in Wave bioreactorupdate on Production of Recombinant Therapeutic ProteincoCHO-DG4422 mg/LMabProcess scaled up to 110 L bioreactor with 80 LEbook Update on production of recombinant therapeutic protein – Transient gene expression: Part 2
working volume[201CHO300 mg/L (at 5 ml scale)Mab1) Reduced pDNA by 50%|6|250 mg/L (at 500 mL scale)2) Transfection at a high density3) Eliminating th3 Optimisation of Transient Gene Expression for Therapeutic Protein ProductionJianwei ZhuTransient gene expression (TGE) is a well-established technol Ebook Update on production of recombinant therapeutic protein – Transient gene expression: Part 2ts: CHO versus HEK293[1241HEK2934-9.1 mg/LCHO-S50-60 mg/LMabFlask at 1 L scale using FreeStyle TM MAX. Fluman erythropoietin and human blood coagulation factor IX were expressed in both HEK293 and CHO cells[1231HEK293F40-50 mg/LCHO-DG4430-60 mg/LMabDisposable shake bioreactor at 30 L working volume. Ebook Update on production of recombinant therapeutic protein – Transient gene expression: Part 2[127]HEK29369-165 mg/LHuman IgG AntibodyLarge scale in Wave bioreactor[131oo-Uc;Ep/'-CHO140 mg/LMabEBVNA1 and oriP in expression plasmid1781HEK293F100Ebook Update on production of recombinant therapeutic protein – Transient gene expression: Part 2
0 nig/LHuman IgGHigh density cell culture using Expi293 Medium[2]CHO63 mg/Lr-proteinAJe/zowrws-based expression at small scale1128]3 Optimisation of Transient Gene Expression for Therapeutic Protein ProductionJianwei ZhuTransient gene expression (TGE) is a well-established technol3 Optimisation of Transient Gene Expression for Therapeutic Protein ProductionJianwei ZhuTransient gene expression (TGE) is a well-established technolGọi ngay
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