Virus purifying and amplification method

Abstract

The invention discloses a virus purifying and amplification method. The method comprises steps: firstly, cell fragments in viruses are removed; secondly, the viruses are captured through a chromatography; thirdly, the viruses captured in the second step are processed by nucleases, host cell nucleic acids in the viruses are subjected to enzymolysis; fourthly, the viruses processed in the third steps are subjected to fined purifying, host cell protein and nucleic acid fragments are removed. The chromatography is employed to achieve virus purifying and amplification, and optimization is carried out at the chromatography column and chromatography filling material aspects. When the provided method is compared with traditional virus purifying method, the purity and the titer of the viruses after purifying are raised, and the provided method has advantages of easy amplification and low cost. The obtained viruses with low residues, high purity and high titer are used for virus removal verification, a flux of a virus filtering membrane can be raised, and the cost investment of the virus filtering membrane is lowered.

Description

technical field [0001] The invention relates to the field of biology, in particular to a method for virus purification and amplification. Background technique [0002] In order to improve the safety of blood products, biological tissue extraction products, and eukaryotic cell expression products, and to prevent possible potential harm to humans in the raw materials of biological products, the production process of biological products needs to include effective removal / inactivation of these potential viruses. Virus processing steps to ensure the biosafety of the product. [0003] To verify whether the virus removal / inactivation process steps in the production process are effective, it is necessary to artificially add a certain type and quantity of indicator virus to the sample. Nano-membrane filtration is a common method in virus removal verification. In order to achieve a sufficient virus removal coefficient and ensure biological safety to pass the national review, a suffic...

AI Technical Summary

Problems solved by technology

Traditional virus purification methods include ultracentrifugation, precipitation, and ultrafiltration. These methods can be used for small-scale virus purification, but due to equipment limitations, the purification efficiency is low, and it is difficult to scale up and purify production, and the virus purity is not very high. Ideal (refer to the Chinese patent application with application number CN201510183392.4 and titled "A Purification Method of Recombinant Herpes Simplex Virus")

For some fragile viruses, such as enveloped viruses, the physical action brought about by ultracentrifugation can even cause damage to the virus structure, resulting in the loss of biological activity of the virus