Philippe Pierre Girard

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Recent advances in genomics, proteomics, and structural biology raised the general need for significant amounts of pure recombinant protein (r-protein). Because of the difficulty in obtaining in some cases proper protein folding in bacteria, several method ...

An agitated 12-well microtiter plate system with a working volume of 2ml was investigated for cell culture process development. Agitation assures homogeneity in wells and enhances mass transfer between the gas and the liquid phase, thus improving maximum cell density and pH stability. The pH of the NaHCO(3)-buffered system can be adjusted by altering the carbon dioxide content of the gas phase. The non-toxic, visual pH indicator phenol red was used in combination with a spectrophotometric plate reader for r

We have developed a novel method called Calfection for gene delivery to and protein expression from suspension-cultivated mammalian cells. Plasmid DNA was simply diluted into a calcium chloride solution and then added to the cell culture for transfection. We evaluated and optimized this approach using suspension-adapted HEK293 cells grown in 12-well plates that were shaken on an orbital shaker. Highest expression levels were obtained when cells were transfected at a density of 5x10(5) cells/ml in the presen

This is the first report of two successful 100 l scale transienttransfections in a standard stirred bioreactor. More than half a gram of a monoclonal antibody (IgG) were produced in less than 10 days using a technology called large-scale transient gene expression(LS-TGE). Suspension adapted HEK 293 EBNA SF cells were transfectedwithin a 150 l (nominal) bioreactor by a modified calcium phosphateco-precipitation method with more than 75 mg of plasmid DNA per run.A mixture of three different plasmids, one enco

To date, methods for large-scale transient gene expression (TGE) in cultivated mammalian cells have focused on two transfection vehicles: polyethylenimine (PEI) and calcium phosphate (CaPi). Both have been shown to result in high transfection efficiencies at scales beyond 10 L. Unfortunately, both approaches yield higher levels of recombinant protein (r-protein) in the presence of serum than in its absence. Since serum is a major cost factor and an obstacle to protein purification, our goal was to develop a

Microinjection is a gene transfer technique enabling partial control of plasmid delivery into the nucleus or cytoplasm of cultured animal cells. Here this method was used to establish various recombinant mammalian cell lines. The injection volume was estim ...

For large-scale applications in biotechnology, cultivation of mammalian cells in suspension is an essential prerequisite. Typically, suspension cultures are grown in glass spinner flasks filled to less than 50% of the nominal volume. We propose a superior ...

Described is method for introducing a desired nucleic acid into a eukaryotic host cell, said method comprising a) admixing the desired nucleic acid, a mono- and/or divalent metal cation and a eukaryotic host cell lacking a cell wallb) incubating the mixture to allow the eukaryotic host cell to take up the nucleic acid.

Philippe Pierre Girard

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Transient gene expression in suspension HEK-293 cells: application to large-scale protein production

Small-scale bioreactor system for process development and optimization

Calfection: a novel gene transfer method for suspension cells

100-liter transient transfection

Serum-free large-scale transient transfection of CHO cells

Generation of recombinant Chinese hamster ovary cell lines by microinjection

Orbital shaker technology for the cultivation of mammalian cells in suspension

A method for introducing a nucleic acid into a eukaryotic cell

Transient gene expression: A novel mammalian cell-based technology for recombinant protein production. A historical and technical perspective

Rapid protein expression in mammalian cells : large-scale transient transfection

Transient gene expression in suspension HEK-293 cells: application to large-scale protein production

Calcium phosphate transfection optimization for serum-free suspension culture

Scalable, Serum-free, High Efficiency Transfection of CHO Cells using an Economical DNA Transfer Vehicle

Gene Transfer to animal cells in culture by microinjection: A novel tool to create recombinant cell lines?

100 Liter Transient Transfection

Tubespin satellites: a fast track approach for process development with animal cells using shaking technology