Can I use antibiotics as a selection marker for Expi293F cell (HEK293F)?

No, antibiotics are typically not used as selection markers for Expi293F cells or HEK293F cells. Instead, these mammalian cell lines are typically selected and maintained using a different method known as antibiotic-free selection or suspension adaptation.

Here's how it works:

Suspension Adaptation: Expi293F cells and HEK293F cells are adapted to suspension culture conditions, which means they can grow and be maintained in culture without attaching to a surface. This is crucial for large-scale protein expression and biomanufacturing applications.

Selection Without Antibiotics: These cells are usually selected and maintained based on their ability to grow in suspension culture rather than relying on antibiotics. Cells that are not adapted to suspension culture conditions will not thrive in this environment and will eventually die off.

Expression Vectors: To express a protein of interest in Expi293F or HEK293F cells, you typically use specialized expression vectors that contain a promoter for driving protein expression (e.g., CMV, EF1α) and, if necessary, a selection marker such as a resistance gene (e.g., neomycin resistance) that allows you to select for cells that have stably integrated the expression vector into their genome.

Transfection: You transfect the cells with the expression vector using a suitable transfection reagent or system. After transfection, you allow the cells to recover and express the protein of interest.

Selection for Stable Expression: If you require stable expression of your protein of interest, you can select for cells that have integrated the expression vector into their genome by applying a selective pressure (e.g., G418 for neomycin resistance) for a period of time. Cells that have successfully integrated the vector will survive and continue to express the protein of interest.

So, in summary, antibiotics are not used as selection markers in Expi293F or HEK293F cells because these cells are primarily adapted to suspension culture conditions, and selection is typically based on the ability to grow in suspension and the use of selection markers within expression vectors, not antibiotics.

Sabine Strehl

The simultaneous application of three antibiotics may be tricky. I would suggest to first conduct kill curve/dose response experiments with each of the antibiotics on the non-transfected cells to determine the optimal/minimum concentration of each antibiotic, which will kill all cells over a specific period of time (e.g. 2-5 days). You might then need to apply all three antibiotics simultaneously to the non-transfected cells at the determined optimal concentrations to adapt the time for the actual selection experiment of the transfected cells.

In what cases low alignment score but high mapping score??

Is there a problem with the presence of metal ions(Zn) in proteins?

Why Do TDS and EC Increase with Larger Wastewater Volumes, While BOD and COD Decrease?

Could it be a cell culture contamination?

Is there any way to quantify bacterial and fungal cells in their mixed culture?

How to normalize and take the significance of the MTT OD values with 3 replicates for the same cell-line?

Why activated CAR-Jurkat cell could not kill targets?

How to isolate lymphocytes from mouse spleen?

How to Add Missing Water Molecules in Protein-Membrane Simulations?

Does long-term culture of murine Monocytes result in macrophage differentiation?

Does matter flow through ecosystems and energy is recycled and how does the flow of matter and energy compare in the living world?

To perform transfection with DharmaFECT Duo in AGS cells. Could you tell me what the ideal concentration is to avoid significant cytotoxicity?