Effect of addition of 'carrier' DNA during transient protein expression in suspension CHO culture.

نویسندگان

  • Ketaki Pradhan
  • Mugdha Gadgil
چکیده

Transient protein expression using polyethyleneimine as a transfection agent is useful for the rapid production of small amounts of recombinant proteins. It is known that an increase in extracellular DNA concentration during transfection can lead to a nonlinear increase in intracellular DNA concentration. We present an approach that hypothesizes that this nonlinearity can be used to decrease the amount of plasmid required for productive transfections. Through addition of non coding 'carrier' DNA to increase total DNA concentration during transfection, we report a statistically significant increase in protein (IgG) expression per unit plasmid used for transfection. This approach could be useful to increase protein yields for large scale transfections under conditions where plasmid availability is limited.

برای دانلود رایگان متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Determination of Minimum Inhibitory Concentration (MIC) of Hygromycin B in CHO cells

Chinese hamster ovary (CHO) cells are considered as the most commonly used host for industrial manufacturing of therapeutic proteins. The aim of this study was to evaluate the minimum inhibitory concentration (MIC) of hygromycin B in both CHO-DG44 and CHO-S cells since hygromycin B resistance cassette can be used for future selection of gene expression in CHO cells. The minimum inhibitory conce...

متن کامل

P-66: Optimization of Human Luteinizing Hormone Expression in CHO Cells Culture by Stepwise Reduction in Serum Concentration

Background: Mammalian Cell lines are the main expression system for the production of recombinant therapeutic proteins. Optimization of cell culture condition is performed via alteration in different parameter. Cell culture media plays an important role in cell cycle because of compounds such as amino acids, vitamins, inorganic salts, glucose, and serum as a source of growth factors, trace elem...

متن کامل

Intracellular Localization of FLAG-Peroxisomal Protein in Chinese Hamster Ovary (CHO) Cells

Epitope tagging is a method of expressing proteins whereby an epitope for a specific monoclonal antibody is fused to a target protein using recombinant DNA techniques. The aim of this study was to sub-clone the peroxisomal protein (PEP) cDNA into a mammalian expression vector leading to the formation of a  chimeric PEP-cDNA containing the FLAG epitope. The FLAG-PEP recombinant cDNA was construc...

متن کامل

[Transient gene expression of soluble VEGFR2: I-IV].

The extracelluar domain I-IV of target gene VEGFR2 (Vascular endothelial growth factor receptor 2) was cloned from villus of trimester abortion by RT-PCR, and linked to the expression vectors. Then, the transfection conditions were optimized in serum-free suspension culture HEK293 using GFP (Green fluorescence protein) as the report gene. The results showed that the optimal transfection efficie...

متن کامل

مقایسه میزان تولید پروتئین نوترکیب در دو رده سلولی CHO و MCF7

Background : The aim of the present study was to identify a suitable cell line for studies of recombinant protein production in eukaryotic system. After transfection, altered expression levels of RNA and its target protein were analyzed. Materials and Methods: To investigate the in vitro expression of E6 protein of human papillomavirus type 16 in cell culture, the plasmid pcDNA3-E6, and two di...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

عنوان ژورنال:
  • Cytotechnology

دوره 64 6  شماره 

صفحات  -

تاریخ انتشار 2012