نتایج جستجو برای: cho dg44
تعداد نتایج: 13351 فیلتر نتایج به سال:
Backround Glycosylation of protein therapeutics is influenced by a multifaceted mix of product intrinsic properties, host cell genetics and upstream process parameters. Industrial CHO cell lines may have several deficits in their glycosylation pattern for some applications, like high fucose content (corresponding to a low ADCC profile) and low galactosylation and sialylation levels (proposed to...
The cDNA or protein of interest, a Human-Fc TF20X40L fusion protein, was synthesized and integrated into a vector containing DHFR gene. This expression vector was then transfected into DG44, a DHFR deficient CHO cell line by electroporation. The transfected pool was selected with gradient MTX pressure before subcloning in semi-solid media. 1500 primary clones were screened for titer by ELISA. T...
To generate industrially applicable new host cell lines for antibody production with optimizing antibody-dependent cellular cytotoxicity (ADCC) we disrupted both FUT8 alleles in a Chinese hamster ovary (CHO)/DG44 cell line by sequential homologous recombination. FUT8 encodes an alpha-1,6-fucosyltransferase that catalyzes the transfer of fucose from GDP-fucose to N-acetylglucosamine (GlcNAc) in ...
Materials and methods CHO-DG44 cells were cultivated in suspension in 1-L bottles as described in [1]. To determine conditions under which the shear stress was harmful for the cells, the bottles were orbitally shaken on an ES-X platform (Kühner AG, Birsfelden, Switzerland) at agitation rates from 150 to 200 rpm for 24 h. Control cultures were run in parallel with agitation at 110 rpm. The veloc...
Background Transposon systems mediate stable integration of exogenous DNA elements into a host cell genome, and have been successfully used in mammalian cells for the generation of stable cell lines. The piggyBac (PB) transposon system has been shown to have several advantages over the other transposon system available [1-3]. It has also been shown to generate stable cell lines at significantly...
BACKGROUND Trastuzumab (Herceptin) is a humanized monoclonal antibody (mAb) which is used for specific treatment of metastatic breast cancer in patients with overexpression of HER2/neu receptor. In this study, we have attempted to develop a biosimilar version of trastuzumab mAb. METHODS According to in silico studies, the heavy and light chains of trastuzumab mAb were designed and constructed...
To date, methods for large-scale transient gene expression (TGE) in cultivated mammalian cells have focused on two transfection vehicles: polyethylenimine (PEI) and calcium phosphate (CaPi). Both have been shown to result in high transfection efficiencies at scales beyond 10 L. Unfortunately, both approaches yield higher levels of recombinant protein (r-protein) in the presence of serum than in...
Background Cell line development (CLD) is a critical step in the generation of biotherapeutics, but it is still hindered by several pain points, including the lengthy and laborintensive workflow needed to isolate desirable clones, lack of reproducibility, as well as potential protein quality issues. Over the last decade, antibody titers in mammalian cell culture systems in excess of 3 g/L have ...
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