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Role of non-specific DNA in reducing coding DNA requirement for transient gene expression with CHO and HEK-293E cells

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The first reports about successfully expressed recombinant proteins with the use of a baculovirus vector were published over 30 years ago. Despite the long time of refining this expression system, early problems with the production of baculovirus-derived s ...

Academic Press Inc Elsevier Science2015

Virus-free transient protein production in Sf9 cells

Florian Maria Wurm, David Hacker, Lucia Baldi Unser, Xiao Shen

A method for virus-free transient gene expression from suspension-adapted Sf9 insect cells was developed with the gene of interest being expressed from a plasmid carrying the homologous region 5 enhancer (hr5) and the immediate early 1 (ie1) promoter from ...

Elsevier2014

Due to the high demand for heterogeneous protein production in insect cells, we have established efficient methods for plasmid-based transient gene expression (TGE) in various insect cell lines, including commonly used Spodoptera cell lines Sf9 and HighFiv ...

EPFL2014

Transient gene expression (TGE) is a rapid method for the production of recombinant proteins. Recently, valproic acid (VPA), a well known histone deacetylase inhibitor, has been suggested for the use in cell cultures to enhance recombinant protein expressi ...

EPFL2013

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Transient gene expression (TGE) from mammalian cells is an increasingly important tool for the rapid production of recombinant proteins for research applications in biochemistry, structural biology, and biomedicine. Here we review methods for the transfect ...

Elsevier2013

Protein tag-mediated conjugation of oligonucleotides to recombinant affinity binders for proximity ligation

Kai Johnsson

While antibodies currently play a dominant role as affinity reagents in biological research and for diagnostics, a broad range of recombinant proteins are emerging as promising alternative affinity reagents in detection assays and quantification. DNA-media ...

Elsevier2013

Reduced glutamine concentration improves protein production in growth-arrested CHO-DG44 and HEK-293E cells

Florian Maria Wurm, David Hacker, Lucia Baldi Unser, Yashas Rajendra, Divor Kiseljak

For most cultivated mammalian cells, glutamine is an essential medium component. However, glutamine consumption results in the production of ammonia, a cytotoxic byproduct. Here we investigated the effect of glutamine reduction on recombinant protein produ ...

Springer Netherlands2012

Hyperbranched Polylysine: A Versatile, Biodegradable Transfection Agent for the Production of Recombinant Proteins by Transient Gene Expression and the Transfection of Primary Cells

Harm-Anton Klok, Florian Maria Wurm, David Hacker, Lucia Baldi Unser, Yashas Rajendra, Zuzana Kadlecova

The feasibility of a new transfection agent, HBPL, for the production of recombinant IgG antibody via TGE as well as for the transfection of primary cells is studied. Under the conditions investigated, transfection of CHO-DG44 cells using HBPL results in I ...

2012

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The large-scale transfection of mammalian cells allows moderate (milligram to gram) amounts of recombinant proteins (r-proteins) to be obtained for fundamental or clinical research. In this article, we describe a one-liter transfection using polyethyleneim ...

Humana Press2011

Gene Delivery with Hyperbranched Polylysine

Zuzana Kadlecova

The delivery of exogenous nucleic acids into mammalian cells is a valuable technique for basic research studies on the expression, regulation and function of genes and proteins. At the same time, gene delivery to cultivated mammalian cells has become funda ...

EPFL2011