A device and method for large-scale continuous multiple suspension transient expression of recombinant proteins

Abstract

The present invention discloses a device and method for expressing recombinant protein by large-scale continuous multiple suspension transient transfection. The device and method mainly utilize perfusion technology and maintain cells in a relatively good growth state, so that the process of transient transfection can be carried out smoothly. Multiple times (≥2), and the cell state of the multiple transient transfection group has no obvious decrease in viability and proliferation during the continuous multiple transient transfection process, so it can effectively extend the cycle of the entire process and increase Increase the yield of recombinant protein and reduce the production cost of recombinant protein.

Description

technical field [0001] The invention relates to a device and method in the field of bioengineering, in particular to a device and method for multiple transient transfection and long-term expression of recombinant proteins in a continuous suspension bioreactor. Background technique [0002] Transfection refers to the process by which eukaryotic cells acquire new genetic markers due to the incorporation of foreign DNA. Conventional transfection techniques can be divided into two categories: transient transfection and stable transfection. The exogenous DNA / RNA is not integrated into the host chromosome during transient transfection, so multiple copy numbers can exist in a host cell, resulting in high-level expression, but usually only for a few days, after 24 hours to 96 hours after transfection Harvest cells within hours, depending on cell type, vector construction, and many other factors. [0003] In recent years, with the development of transient transfection technology an...

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Problems solved by technology

[0005] First, due to the characteristic of plasmid loss during transient transfection, transient transfection technology is limited to the laboratory level, and industrialization has many limitations;

[0006] Second, the common large-scale transient transfection process (such as batch) generally lasts 3-7 days, and the cycle is short, resulting in high cost of the entire process

[0007] To sum up, the existing method of transient gene transfection to express recombinant protein has short period, low yield and high cost, and is not suitable for industrial-scale production, and it is difficult to meet the demand for recombinant protein in preclinical experiments

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