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Rational vector design and multi-pathway modulation of HEK 293E cells yield recombinant antibody titers exceeding 1 g/l by transient transfection under serum-free conditions
TLDR
This protocol was reproducibly scaled-up to a working volume of 2 l, thus delivering >1 g of purified protein just 2 weeks after transfection, which is the fastest approach to gram quantities of protein ever reported from cultivated mammalian cells.
High-density transfection with HEK-293 cells allows doubling of transient titers and removes need for a priori DNA complex formation with PEI.
TLDR
Successful in situ transfection of suspension-adapted HEK-293 cells with 25-kDa linear polyethyleneimine at densities up to 20 x 10(6) cells/mL in complex media followed by production at lower cell densities is reported.
Valproic acid: a viable alternative to sodium butyrate for enhancing protein expression in mammalian cell cultures.
TLDR
With valproic acid (VPA), volumetric and specific productivities of 200 mg/L and 20 pg/cell/day, respectively, were achieved in HEK293E cells with a 10-day process and are recommended as a cost-effective alternative to this widely used enhancer of recombinant protein production from mammalian cells.
High Cell Density Transient Gene Expression in HEK 293 EBNA Cells
TLDR
Results obtained by optimization of a PEI-mediated high-density transient transfection protocol in HEK-293E cells with emphasis on reducing the cost of DNA needed for transfections are presented.
A Transient Gene Expression Process with Recombinant Antibody Titer of 0.5 g/L in CHO Cells
TLDR
An optimized transfection method for Chinese hamster ovary (CHO) cells is described that allows the transient production of recombinant antibody at titers approaching 0.5 g/L by two weeks post-transfection.
Release of Plasmid-DNC from PEI/DNA Particles
TLDR
The results of these in vitro experiments are important for understanding the transport and release of plasmid DNA from PEI/DNA particles after transfection of cells.
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