Method for extracting, concentrating and purifying porcine senecavirus particles by two aqueous phases

Abstract

The invention belongs to the technical field of two-aqueous-phase extraction of viruses, and discloses a method for concentrating and purifying porcine SenecaviusA (SVA) particles through two-aqueous-phase extraction. The method comprises the following steps: performing SVA reproduction; carrying out indirect immunofluorescence identification on the SVA; establishing a three-step aqueous two-phase extraction method; determining the content of SVA; carrying out a western blot test; and carrying out electron microscope identification on SVA. After low-concentration polyethylene glycol and inorganic salt are added into an SVA culture solution, cell debris is removed, and then polyethylene glycol is added to form an aqueous two-phase system, so that concentrated and purified porcine SVA virus particles can be quickly obtained; rapid separation of virus particles can be realized through an aqueous two-phase extraction technology, the recovery rate and the purity of the virus particles are relatively high, the process is simple, and the cost is low; through a three-step aqueous two-phase extraction method, the purpose of concentrating and purifying the porcine SVA virus particles from the cell culture fluid is achieved, and the method can be applied to laboratory and large-scale production at the same time.

Description

technical field [0001] The invention belongs to the technical field of virus two-phase extraction, in particular to a method for concentrating and purifying porcine Seneca virus particles through two-phase extraction. Background technique [0002] At present, the two-phase extraction technology is a technology that uses the distribution difference of substances in two immiscible solvents to separate, but its principle is still unclear. When the properties of the extraction system are different, after the substance enters the two-phase system, due to the existence of surface properties, charge effects and various forces (such as hydrophobic bonds, hydrogen bonds and ionic bonds, etc.) The concentration in the upper and lower phases is different, so as to realize the separation of the target substance. As a new separation and purification method, two-phase extraction technology is widely used in the separation of polypeptides, proteins, enzymes, nucleic acids, viruses, cells,...

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Problems solved by technology

[0008] (2) The purity of PEG-precipitated virus is very high, but the virus recovery rate is low, and the sample concentration and purification time is long, and it needs to cooperate with centrifugation and other technologies, and also requires professional equipment

The loss of virus easily caused by PEG purification is unfavorable for the reduction of cost for cheap animal vaccines

[0009] The difficulty in solving the above problems and defects is: the inherent characteristics of ultrafiltration and PEG purification not only limit their application in large-scale production of animal vaccines, but also are not conducive to the improvement of vaccine quality

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