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Concept

Asilomar Conference on Recombinant DNA

Related publications (22)

The enhancement of DNA fragmentation in a bench top ultrasonic water bath with needle-induced air bubbles: Simulation and experimental investigation

Martinus Adela Maria Gijs, Thomas Lehnert

Shearing DNA to a certain size is the first step in many medical and biological applications, especially in next-generation gene sequencing technology. In this article, we introduced a highly efficient ultrasonic DNA fragmentation method enhanced by needle ...

AIP Publishing2022

Single Molecule Localization and Discrimination of DNA–Protein Complexes by Controlled Translocation Through Nanocapillaries

Aleksandra Radenovic, Sebastian Maerkl, Sanjin Marion, Roman Bulushev, Sebastian James Davis, Ekaterina Emilova Petrova

Through the use of optical tweezers we performed controlled translocations of DNA-protein complexes through nanocapillaries. We used RNA polymerase (RNAP) with two binding sites on a 7.2 kbp DNA fragment and a dCas9 protein tailored to have five binding si ...

American Chemical Society (ACS)2016

Study of transposon-mediated cell pool and cell line generation in CHO cells

Sowmya Balasubramanian

The goal of this thesis was to evaluate the use of artificial transposon systems for the generation of recombinant cell pools and cell lines with Chinese hamster ovary (CHO) cells as the production host. Transposons are naturally occurring genetic elements ...

EPFL2015

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

Development of plasmid-based expression technology for rapid and sustained production of recombinant proteins in insect cells

Xiao Shen

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

Mathematical models and computational methods for the analysis of genome-scale protein synthesis

Julien Racle

Proteins are a ubiquitous and indispensable element for every living organism, from simple bacteria to mammals. Already in the simplest organisms, there exist some thousands of different protein species that take up a great variety of structures, and thus ...

EPFL2014

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

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

The Use of Valproic Acid in Transient Gene Expression with HEK-293E Cells

Divor Kiseljak

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

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

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

Transient gene expression (TGE) is a rapid method for the production of recombinant proteins in mammalian cells. While the TGE volumetric productivity has improved significantly over the past decade, the amount of plasmid DNA (pDNA) needed for transfection ...

2012

A simple high-yielding process for transient gene expression in CHO cells

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

Here we describe a simplified method for transient gene expression (TGE) in suspension-adapted Chinese hamster ovary (CHO) cells using polyethylenimine (PEI) for DNA delivery. Both the transfection and production phases of the bioprocess were performed at ...

Elsevier2011

Large-Scale Transfection of Mammalian Cells

Florian Maria Wurm, David Hacker, Lucia Baldi Unser

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

The PiggyBac transposon enhances the frequency of CHO stable cell line generation and yields recombinant lines with superior productivity and stability

Florian Maria Wurm, David Hacker, Lucia Baldi Unser

Generating stable, high-producing mammalian cell lines is a major bottleneck in the manufacture of recombinant therapeutic proteins. Conventional gene transfer methods for cell line generation rely on random plasmid integration, resulting in unpredictable ...

Wiley-Blackwell2011

Transient Gene Expression in HEK-293E Cells for Recombinant Protein Production : Process Characterization towards GMP Approval

The growing demand of biopharmaceutical products is boosting the market for therapeutic recombinant proteins (r-proteins). More than half of the 140 r-proteins that have gained approval for human therapeutic use are manufactured in mammalian cells that mak ...

EPFL2010

Transient recombinant protein expression in mammalian cells : the role of mRNA level and stability

Sarah Wulhfard

Transient gene expression (TGE) is a rapid method for generating recombinant proteins in mammalian cells, but the volumetric productivities for secreted proteins in transiently transfected CHO DG44 cells are typically more than an order of magnitude lower ...

EPFL2009

Accelerating the development time of recombinant mammalian cell lines producing high titers of protein therapeutics

Sébastien Chenuet

The forthcoming arrival on the market of numerous protein therapeutics that require high clinical doses will foster the need for high-producing mammalian cell lines. Furthermore, pressures to reduce the development time for new therapeutics has meant more ...

EPFL2008

Development of high-titer transient gene expression processes for manufacturing recombinant proteins

Gaurav Backliwal

The market for recombinant therapeutic proteins (including antibodies) is estimated to be greater than $50-billion and has a compounded annual growth rate of around 20%. More than 50% of all approved processes for manufacturing recombinant proteins use mam ...

EPFL2008

The use of RNA interference to increase PEI-mediated transient gene expression in mammalian cells

Martin Bertschinger

DNA uptake by polyethylenimine (PEI)-mediated transfection was investigated in Chinese hamster ovary (CHO DG44) cells. Rapid DNA uptake was observed with the maximum occurring during the first 45-60 min after addition of PEI-DNA complexes to cells. With lo ...

EPFL2006

Establishment of recombinant mammalian cell lines : new approaches

Madiha Derouazi

Recombinant proteins are gaining in importance for therapeutic applications. The proteins are expressed in stable cell lines, within which recombinant DNA has incorporated into the host cells genome. Identification and isolation of extremely high producers ...

EPFL2005