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Direct Procurement of ReAgent 007, a Transient Transfection Reagent for a High-throughput Midscale Protein Production in Chinese Hamster Ovary (CHO) Cells

The Human Secretome project within the Wallenberg Center for Protein Research (WCPR) at the Royal Institute of Technology (KTH) has set up a high-throughput protein production pipeline to be able to produce a large number of recombinant proteins in CHO cells yearly. Now the production will change from small scale to midscale to produce higher amounts of each protein. To be able to achieve this a chemical transient transfection reagent is needed and for that the reagent “ReAgent 007” is to be purchased.

It is an expression system that uses specific mammalian cells (QMCF cell lines) and vectors (QMCF plasmids) for production of recombinant proteins. We have established a robust production pipeline with this technology and we will now shift the production from small scale to midscale using the same cells and vectors. During the last year we have evaluated different protocols, testing culture media, feed media, shake flasks and transfection reagents for the upscaled production. The chosen protocol, for maximum transfection efficiency in the Icosagen system includes a transfection reagent that is produced and distributed by Icosagen Cell Factory. ReAgent007, the transfection reagent chosen, was the only one fulfilling our demands. For example, resulting in the highest transfection efficiency leading to high enough protein amounts in a minimum culture volume and thereby the upscaled production can be performed in our already existing laboratory equipment.

The Human Secretome project within the WCPR at KTH has since the start of the project (2013) been using a technology called the QMCF Technology (Patent number: EP1851319) developed by the Estonian company Icosagen Cell Factory OÜ. It is an expression system that uses specific mammalian cells (QMCF cell lines) and vectors (QMCF plasmids) for production of recombinant proteins and has been chosen after extensive testing of different expression systems. Today we have generated over 3,000 expression vectors that all are designed to work exclusively in Icosagen QMCF cells. Next step for our production is to produce higher amounts of each protein using these cells and vectors and still keeping a high-throughput pipeline. During the last year one of the researchers in the ProteinFactory group has evaluated different protocols. For example, different culture media, feed media, shake flasks and transfection reagents have been evaluated for the upscaled production. The optimized protocol now chosen for maintaining a high-transfected cell density, viability, and to achieve the highest titers in the Icosagen system includes a transfection reagent that is produced and distributed by Icosagen Cell factory. This new cultivation protocol also works in a high-throughput way that is necessary for our production pipeline.

Because of the above stated reasons it is crucial for our upscaled production that we are able to buy this transfection reagent from the Icosagen Cell Factory Company that is the manufacturer of this transfection reagent and the only company that sells this product.

Visma notice: https://opic.com/id/afqppfmcjf