Insect cell baculovirus vector expression system is widely used in the production of recombinant proteins. Compared with typical E. coli and yeast expression systems, it has a powerful polyhedrin promoter, so it has high expression performance and can perform correctly. The post-translational modification to produce functional and immunogenetic active recombinant protein. Moreover, the virus and parasite proteins produced by the baculovirus vector expression system have been tested to have good immunogenicity.
Insect baculovirus can be expressed not only in insect culture cells, but also in insects and pupae. For conventional protein production in small quantities and with high purity, the use of in vitro cultured insect cells for expression is the first choice. The cell lines used in the insect cell baculovirus vector expression system are mainly derived from lepidopteran insect cell lines, especially Bombyx mori, Mamestra brassicae, Spodoptera frugiperda and Anopheles mosquitoes The cell line of Trichoplusia ni, among which Sf9 cells, Sf21 cells, Tn-368 cells and Hign Five cells are the most widely used. Medicilon researchers have established a mature baculovirus-insect cell expression service platform, providing services including the preparation of recombinant baculovirus, the expression and purification of recombinant proteins and their complexes.
Since 1983, researchers have successfully expressed human interferon p in Spodoptera frugiperda Laphygmafrug true Ferda cell Sf9 using AcNPV (AcNPV). In 1985, Maeda used Bombyx mori Bo-m61zmD/ g NPV is used as an expression vector to express human alpha-interferon at a high level in silkworms. As one of the four major expression systems of genetic engineering today, the baculovirus vector expression system has become increasingly popular due to its unique advantages and great development potential. Scholars from various countries pay attention to it. The commonly used cell lines of insect cell baculovirus vector expression system are as follows:
Sf21 cells are derived from the ovarian tissue of female Spodoptera frugiperda pupae, and are a clone of Sf9 cells; Sf21 cells can be used for basic research on insect viruses and for replicating baculovirus expression vectors. The research object of the Sf21 cell line is living cells. During the experiment, it can always maintain cell viability according to requirements, and can monitor, detect and even quantitatively evaluate the condition of some living cells, including the morphology, structure, and life activities of living cells. . And the research conditions can be controlled artificially, such as pH, temperature, oxygen, carbon dioxide, tension and other physical and chemical conditions, which can be controlled artificially according to actual needs. At the same time, chemical, physical, biological and other factors can be used as conditions for experimental observation, and these factors can also be under strict control.
Sf21 cell line research samples can achieve relatively uniformity. After a certain number of generations of cell culture, the obtained cell line can achieve homogeneity and belong to the same type of cells. If necessary, cloning and other methods can be used to purify and study the cells. The content is easy to observe, detect and record. From lower animals to higher animals, different age stages and different tissues of an animal can be targeted for research.
Sf9 cells are derived from the ovarian tissue of female Spodoptera frugiperda pupa, which can be used to replicate baculovirus expression vectors. It is a branch of Sf21 cells. Both Sf9 cells and Sf21 cells are derived from the ovarian cells of Spodoptera frugiperda. Sf21 is the cell line used to produce the prostate cancer vaccine Provenge, which has been approved by the FDA for use. Sf9 is a clone of Sf21. Compared with Sf21, Sf9 has better tolerance to PH, osmotic pressure and shear force, so it gradually replaced the use of Sf21. Sf21 and Sf9 cell lines are suitable for baculovirus infection, purification, high-titer virus production and recombinant protein expression. The Sf9 cell line is a suitable host for recombinant protein expression using the baculovirus expression system. Although in the past, people used a static culture system consisting of a culture flask and a serum-containing basal medium to cultivate insect cells, but generally speaking, insect cells did not Adherence dependent, convenient for suspension culture.
The growth and infection characteristics of S9 cells are destined to perform well in the transfection and amplification of baculovirus. However, in terms of recombinant protein expression, the Hign Five cell line derived from the ovarian cells of Spodoptera exigua expresses a higher level of expression. These proteins are even more than 20 times higher than those expressed by Sf9 cells. Moreover, Hign Five’s glycosylation and phosphorylation are more complex than Sf9, so it is more suitable for expressing some secreted recombinant proteins. The virus-like particle vaccine of human papillomavirus is produced in Hign Five cells.
Both Tn-368 and Hign Five are derived from cell lines of Tn. However, another study found that Hign Five cells can be subtly infected by nodavirus, and Hign Five cells can be infected by baculovirus. Stimulates the production of noda virus virions. This discovery raises questions about the safety of cell banks infected with occasional pathogens, so researchers need to adopt more rigorous product purification and description methods. Both Hign Five cells and Sf9 cells have the dual characteristics of adherence and suspension growth, so both can be cultured on a large scale through shake flasks and bioreactors.
The baculovirus vector expression system is an expression system that uses insect baculovirus as a foreign gene vector and insects and insect cells as receptors. Compared with bacterial, yeast and mammalian cell expression systems, it has a high level of expression, the expression product can be post-translationally processed, and its antigenicity, immunogenicity and function are similar to natural proteins, and it can be infected by insect larvae. To achieve large-scale low-cost production of genetic engineering products, the baculovirus vector expression system has broad development prospects.
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