Purification method of recombinant viruses with fluorescent markers

Abstract

The invention discloses a purification method of recombinant viruses with fluorescent markers. The purification method comprises the following steps: 1, goat pox viruses with KLP2 genes being lost arerecombined, specifically, (1) sheep testicular primary cells are cultured, a cell plate is spread, after cell culture reaches 80% or so, the cells are infected through goat pox virus vaccine strains,and (2) virus recombination is conducted; and 2, the recombinant viruses are purified, specifically, (1) the recombinant viruses with fluorescence are picked, (2) the viruses are purified, (3) purification is continued, specifically, the viruses, with the fluorescence, picked in the previous step are repeatedly subjected to freeze thawing, then according to operation of (2) in the step 2, dilution and inoculation are conducted, then picking is conducted, and generally after 3-4 generations, the purified recombinant viruses can be obtained.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for purifying recombinant viruses with fluorescent labels. Background technique [0002] Viruses, whether in natural or recombinant form, find use in the fields of vaccination and gene therapy. Many viruses or virus-like particles can safely and efficiently multiply in cells. Recombinant adenoviruses are a preferred class of viral vectors in gene therapy and immunization. After the virus has propagated in the cell, for almost all applications, the virus must be purified before further use. In some industrial processes for purifying viruses, there is still a need for alternative and preferably improved methods for the purification of fluorescently labeled recombinant viruses. [0003] At present, there are many methods for virus purification, but each has advantages and disadvantages. Flow cytometry screening method, although the purification speed is fast, requires a large...

AI Technical Summary

Problems solved by technology

Flow cytometry screening method, although the purification speed is fast, requires a large number of sample cells, and is also limited by the size of its cell particles and expensive instruments; although the LacZ reporter gene plaque screening method can obtain the target from a small amount of recombinant virus The virus is further purified, but the color development is not obvious, the false positive rate is extremely high, and the purification efficiency is not high; although the fluorescently labeled recombinant virus can clearly see the fluorescence of the recombinant virus band, if it is not fixed or only fixed pick, then it takes a long time to get purified virus

Images