Method for improving transient transfection of insect cell and stably expressing protein expression quantity

Abstract

The invention discloses a method for improving transient transfection of insect cell and stably expressing protein expression quantity. A traditional insect cell cultivation method adopts a phosphate buffer system mostly, CO2 is not required to add during the cultivation process, and the expression quantity of a target protein is relatively low. The method creatively finds that the protein expression quantity can be improved by adding CO2 in a cultivating box or changing an air permeable cover to be a sealing cover after transfection to increase CO2 concentration in a bottle upon the self-breathing effect of insect cells regardless of transient transfection or protein expression and production process of a stable cell pond when the insect cell is cultivated; moreover, the lifting effect is significant. The CO2 is added in the cultivation box during the insect cell cultivation process, or the cultivation bottle cover is changed to be a sealing cover, the method is simple and easy to practice, and the cost is low; the method is extensive in use and capable of significantly improving the protein expression quantity of the insect cells.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to a method for increasing the expression level of transiently transfected and stably expressed proteins in insect cells. Background technique [0002] The insect cell expression system has been used for the expression of many human recombinant proteins including tissue factor and antibodies (Kost et al., 1997). At present, the insect expression system is mainly used to express subunit vaccines. In addition to veterinary vaccines, it is also used to produce some human vaccines and recombinant viruses (Maranga et al., 2002; Ryan and Walsh et al., 2012; Yang et al., 2013; Miller et al., 2012) . Drosophila Schneider2 (Drosophila Schneider2) cells and Spodoptera frugiperda Sf9 (Spodoptera frugiperda) cells are insect cells commonly used for heterologous gene expression, and recombinant protein expression can be performed by constructing a stable cell pool. In recent years, insect cells h...

AI Technical Summary

Problems solved by technology

In recent years, the transient transfection of S2, Sf9 and other cells has begun to use low-cost cationic polymer transfection reagents, such as polyethyleneimine, and recombinant proteins can also be obtained in a short period of time, but its yield needs to be improved urgently

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