Nucleofection is superior to lipofection for transient protein production
In a comparative study the Nucleofector technology (amaxa) and the lipofection reagent Lipofectamine™ 2000 (Invitrogen) have been tested in regard to their suitability for transient protein production. Nucleofection achieves 10-30 times higher expression rates in CHO cells compared to cells transfected with lipofection. A similar ratio has been determined for both the specific and the volumetric productivities.
Experimental setup:
2 different suspension CHO cell clones have been transfected with a plasmid encoding the human secreted alkaline phosphatase (SEAP) either using Lipofectamine™ 2000 or nucleofection.
Transfection conditions:
Post transfection cells were cultivated as batches in spinner flasks containing 50 ml culture medium for 5 days under non-regulated conditions.
Suspension 293 cells (DSMZ, adapted from adherent cell clone) have also been tested successfully. 5 x 107 (= 5 samples) cells have been transfected with 25 μg pDrivehSEAP using Cell Line Nucleofector Kit V in combination with program X-001. A specific productivity of 2.4 µU/c/d and a volumetric productivity of 10.5 U/ml could be obtained. These cells were not transfectable using lipofection.
Nucleofection is superior to lipofection for transient protein production.
Specific productivities (µUnits/cell/day) of hSEAP referred to a cell density of 106 cells/ml seeded after the respective transfection.
