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Modified atmosphere: enhancing insect cell cultures experimentation

Modified atmosphere: enhancing insect cell cultures experimentation
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Modified atmosphere: enhancing insect cell cultures experimentation - 1

High Performance Gas flow Dilutor & Gas flow Mixing System with User Interface Application Note MCQ Gas Blender 100 Series Application Modified atmosphere: enhancing insect cell cultures experimentation Introduction Vaccines, antibiotics an others essential medicines have always constituted basic necessities for our society. The need of these goods, greatly increased over the last several decades, leads the modern science towards new reliable and efficient drug production methods. The large-scale production of basic medicines relies on the capacity to synthesize adequate amounts of specific proteins, naturally expressed in their wild type form or appositely modified. One of the most widely used technique designed to enhance the proteins production, is the Heterologous Protein Expression (HPE). HPE technique is base on the following principles: The cell type that naturally expresses the target protein is selected. The genetic material required to express the target protein is collected from the cell. The genetic material is experimentally transferred into a another cell type that does not normally express that protein. The cell type with the new inserted genetic material gains the ability to express the target protein. The two cell types usually derive from different organisms, thus the term heterologous. The expression of functional proteins in heterologous hosts is currently a cornerstone of modern biotechnology. Aside from the large-scale commercial applications, this techniques is an incredible useful tool for lab basic research, thus requiring constant improvements in speed, costs and effectiveness. A key step of the proteins production is the cell culture, during which the system atmosphere plays a crucial role. For this reason the MCQ proposes its Gas Blender 100 Series as a fundamental tool to control, study and improve the culture expression. www.mcqinst.com Gas Blender 100 Series Application Note

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Modified atmosphere: enhancing insect cell cultures experimentation - 2

High Performance Gas flow Dilutor & Gas flow Mixing System with User Interface Baculovirus Expression Vector System The modern HPE exploits many types of heterologous hosts. The most widely used expression systems are based on E. coli as the host cell. Yeats, molds and mushrooms (i.e. fungi) are another important class of hosts (more detailed information about fungi cultures can be found here [link]). A relatively new and promising approach to the expression system is instead based on insect cell hosts. Like bacteria and fungi, the expression capacity of insect cells is artificially...

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Modified atmosphere: enhancing insect cell cultures experimentation - 3

High Performance Gas flow Dilutor & Gas flow Mixing System with User Interface The AcNPV is known to infect only few lepidopteran (moth) families, and has been proved to be not infectious for mammalian cell lines. The use of this specific baculovirus in therefore not only highly selective for insect cell lines but also safe for humans. Heterologous Host Cell Despite these potential advantages, BEVS suffers some of the peculiar downsides shared by any heterologous expression system. Differences in proteins expressed by mammalian and baculovirus infected insect cells have been described, and...

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Modified atmosphere: enhancing insect cell cultures experimentation - 4

High Performance Gas flow Dilutor & Gas flow Mixing System with User Interface contact with a pure oxygen atmosphere, the saturation value (i.e. the amount of dissolved oxygen) at 25°C and 1 bar is about 8 mg/l. Since the relation between atmospheric oxygen and dissolved oxygen is directly proportional, when pure oxygen is replaced by a lower oxygen atmosphere, the DO decreases accordingly. In case of pure water being in contact with air (21% of oxygen) the dissolved oxygen results in 21% of the saturation value (so 21% of 8 mg/l, about 1,7 mg/l). The influence of dissolved oxygen on insect...

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Modified atmosphere: enhancing insect cell cultures experimentation - 5

High Performance Gas flow Dilutor & Gas flow Mixing System with User Interface Hardware configuration An example of MCQ Gas Blender 100 Series hardware configuration is represented in the image alongside. The instrument works with dry, non-aggressive gases. The gas sources can be both pure or mixtures (in our example pure gases have been chosen for simplicity). The gas cylinders are connected to the instrument through 6 mm diameter tubes and a check valve is installed along each line as backflow prevention device. Each gas media is connected and controlled by a dedicated channel of the Gas...

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