Insect cell maintenance liquid for increasing expression amount of hog cholera virus E2 recombinant proteins and preparation method thereof

Abstract

The invention belongs to the technical field of biology, particularly relates to an insect cell maintenance liquid for increasing the expression amount of hog cholera virus E2 recombinant proteins and a preparation method thereof. To be specific, baculovirus is specifically added in an equilibrium liquid-phosphate buffer solution(1 * PBS) to synthesize the limiting amino acids of E2 proteins, dextran sulfate sodium and polyether F-68 which are used as the maintenance liquid to replace an insect cell cell culture medium used for inoculating recombinant baculovirus. With the same culture conditions, same cell quantity and same virus-inoculating quantity, compared with the insect cell culture medium, the insect cell maintenance liquid for increasing the expression amount of hog cholera virus E2 recombinant proteins increases the infection of cells within an unit quantity, the production quantity of the recombinant baculovirus and the secretion amount of the E2 recombinant proteins.

Description

Technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to an insect cell maintenance solution for increasing the expression of the swine fever virus E2 recombinant protein and a preparation method thereof. Background technique [0002] The baculovirus expression system can provide a good environment for the correct folding of highly expressed foreign proteins in the cell, the matching of disulfide bonds and the formation of oligomers, and the structure and function of the expression product can be close to the natural protein, so The use of insect cell-baculovirus expression system recombinant protein is one of the important directions of subunit vaccine research and development. However, due to the high price of insect cell culture media and the low expression of recombinant proteins, many recombinant baculovirus subunit vaccines have stopped in the laboratory development stage. The conventional insect cell-baculovirus expression sy...

AI Technical Summary

Problems solved by technology

This method does not consider the requirements of cell growth after inoculation and the limiting factors of virus replication, which not only affects the expression of CSFV E2 recombinant protein, but also has high antigen production costs per unit volume

[0003] Current studies have shown that the reasons may be: 1. With the proliferation of insect cells, the demand for energy and limiting amino acids increases, and at the same time, a large amount of cell metabolites (lactic acid, alanine, uric acid, NH 4+ etc.), the pH and osmotic pressure of the culture medium change accordingly, which greatly affects the growth of insect cells and the proliferation of viruses after inoculation.

When the cell culture system lacks glutamine, it will affect the synthesis of nucleic acid and protein by the cells, which will affect the growth of the cells

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