Transfection Reagents


Polytheragene has developed PTG1, a transfection reagent that enhances two of the major limitations encountered with currently available products (low efficiency and high cytotoxicity). 

Main avantages of PTG1

  • Easy and fast transfection method
  • High transfection efficiency on many tested cell lines
  • Longer contact time possible with cells, allowing high transfection efficiency.
  • Possible “reverse transfection” experiments without cytotoxicity, leading to transfect cell ratios in the 70 %-90 % range, depending on the cell lines.

Comparison between Forward (F) and Reverse (R) Transfection with PTG1 and a plasmid DNA encoding eGFP on CHO and HEK293 cells. 

Transfection efficiency of primary culture of rat tenocytes with PTG1 and a plasmid DNA encoding eGFP.

PTG1 induces very low cytotoxicity.

Cells were incubated with increasing quantities of PTG1. Cytotoxicity was measured using MTT assays.
  1. HeLa: Human epithelial ovarian carcinoma cells
  2. HEK293: Human embryon kidney cells
  3. C2C12: murine skeletal muscle cells
  4. ΣCFTE29o-: Human tracheal epithelial cells
*Note that systemic toxicity by intravenous administration in mice (DL50) is about 17.5 mg/kg.

 

Production of recombinant proteins and lentiviral particles

 

 

 

PTG1 and LENTIVIRAL PARTICLE PRODUCTION

HEK293T (1 000 000 cells) were transfected with 5 μg of a pDNA mixture (pTRIP-ΔU3-CMV-EGFP, pCVM D8.91 and VSV-G) either

with PTG1, bPEI or CaP. Fluorescence microscopy analysis of cell cultures after 32h.

 

 

Transfection of cells (2 000 000 HEK293T cells) in suspension was performed with PTG1 in OptiMEM without serum. Supernatants were collected at 16h, 20h and 24 h post-transfection and fresh culture medium was added. Supernatants were collected for a second time after 24 h.

Supernatants were titrated for viral particles production.

 

 PTG1 and RECOMBINANT PROTEIN PRODUCTION

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Transfection of 100 ml of CHO-S cells (1 000 000 cellules/ml) in  OPTIMEM medium using PTG1 or Lipofectamine2000 (LFM)