David Hacker

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Transcriptional and post-transcriptional limitations of high-yielding, PEI-mediated transient transfection with CHO and HEK-293E cells

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

Transient gene expression (TGE) in human embryonic kidney (HEK-293) and Chinese hamster ovary (CHO) cells is a well-established technology for the rapid generation of recombinant proteins. Although the maximum TGE yields have reached 1 g/L or more, the amo ...

American Chemical Society2015

Production of active glycosylation-deficient gamma-secretase complex for crystallization studies

Florian Maria Wurm, David Hacker, Patrick Fraering, Virginie Sandra Braman, Mitko Dimitrov, Hermeto Gerber, Andres Ricardo Lopez Robles

Alzheimer's disease (AD)-associated gamma-secretase is a ubiquitously expressed multi-subunit protease complex embedded in the lipid bilayer of cellular compartments including endosomes and the plasma membrane. Although gamma-secretase is of crucial intere ...

Wiley-Blackwell2015

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

Highly efficient production of the Alzheimer's γ-Secretase integral membrane protease complex by a multi-gene stable integration approach

Florian Maria Wurm, David Hacker, Lorène Aeschbach, Patrick Fraering, Mitko Dimitrov, Sowmya Balasubramanian

Inefficient production of membrane-embedded multi-protein complexes by conventional methods has largely prevented the generation of high-resolution structural information and the performance of high-throughput drug discovery screens for this class of prote ...

2013

Coordinated Effects of Sequence Variation on DNA Binding, Chromatin Structure, and Transcription

Bart Deplancke, David Hacker, Gilles Udin, Sebastian Martin Waszak, Sunil Kumar Raghav, Nouria Hernandez, Alisa Yurovsky, Julien Bryois, Sarah Thurnheer

DNA sequence variation has been associated with quantitative changes in molecular phenotypes such as gene expression, but its impact on chromatin states is poorly characterized. To understand the interplay between chromatin and genetic control of gene regu ...

American Association for the Advancement of Science2013

Disposable 600-mL orbitally shaken bioreactor for mammalian cell cultivation in suspension

Florian Maria Wurm, David Hacker, Lucia Baldi Unser, Dominique Terry Monteil Jr, Saroj Ghimire, Giulia Tontodonati

Orbitally shaken bioreactors (OSRs) support the efficient cultivation of mammalian cells in suspension at various scales of operation. Here we introduce a disposable culture vessel with a ventilated cap having a nominal volume of 600 mL (TubeSpin (R) biore ...

Elsevier2013

Helical-Track Bioreactors for Bacterial, Mammalian and Insect Cell Cultures

Florian Maria Wurm, David Hacker, Lucia Baldi Unser, Stéphanie Tissot, Clara Joséphine Douet

We investigated the cultivation of bacterial, mammalian and insect cells in an orbitally-shaken 250-mL disposable tube that incorporates a helical track (HT) on its inside wall. The mass transfer coefﬁcient of oxygen (kLa) was 200%–400% higher in the HT tu ...

2013

Numerical simulation of orbitally shaken viscous fluids with free surface

Alfio Quarteroni, Florian Maria Wurm, David Hacker, Marco Discacciati, Stéphanie Tissot, Samuel Quinodoz

Orbitally shaken bioreactors are an emerging alternative to stirred-tank bioreactors for large-scale mammalian cell culture, but their fluid dynamics is still not well defined. Among the theoretical and practical issues that remain to be resolved, the char ...

Wiley-Blackwell2013

Study of the improved Sf9 transient gene expression process

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

Insect cells have been widely used for the production of recombinant proteins using recombinant baculovirus for gene delivery [1]. To simplify protein production in insect cells, we have previously described a method, based on transient gene expression (TG ...

2013

Polyethyleneimine-based transient gene expression processes for suspension-adapted HEK-293E and CHO-DG44 cells

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

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

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Highly efficient production of the Alzheimer's γ-Secretase integral membrane protease complex by a multi-gene stable integration approach

Florian Maria Wurm, David Hacker, Lorène Aeschbach, Patrick Fraering, Mitko Dimitrov, Sowmya Balasubramanian

2013

Helical-Track Bioreactors for Bacterial, Mammalian and Insect Cell Cultures

Florian Maria Wurm, David Hacker, Lucia Baldi Unser, Stéphanie Tissot, Clara Joséphine Douet

2013

Transcriptional and post-transcriptional limitations of high-yielding, PEI-mediated transient transfection with CHO and HEK-293E cells

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

American Chemical Society2015