Purification method for split influenza virus vaccine

Abstract

The invention provides a purification method for split influenza virus vaccine. An influenza virus strain is inoculated onto a chick embryo and cultured to obtain a virus solution; the virus solution is sequentially subjected to inactivation and ultrafiltration concentration to obtain an ultrafiltrate; the obtained ultrafiltrate is subjected to cane sugar density gradient centrifugation by using a KII continuous flow centrifuge to obtain an ultra centrifugal solution; the ultra centrifugal solution is subjected to ultrafiltration dialysis for cane sugar removal and molecular sieve gel chromatography to obtain a virus purification solution; the obtained virus purification solution is subjected to virus split by using a split agent TritonX-100 and sodium deoxycholate; after the split is over, the split agent is removed via ultrafiltration dialysis; impure protein is centrifugally removed from the obtained split solution; supernatant is collected, filtered and sterilized so as to obtain the purified influenza virus vaccine primary solution. The purification method is simple and convenient to operate, high in centrifugation capacity and suitable for large-scale production; by adopting a dual-split agent and a centrifugal process, the finished product is greatly improved in activity (hemagglutinin content/protein total content), and approaches the activity ratio of subunit vaccine, as a result, the high-grade split influenza virus vaccine product is obtained.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a purification method of influenza virus split vaccine. Background technique [0002] The purification process of influenza virus split vaccine is to remove most of the impurity proteins and obtain influenza virus after the inactivated influenza virus harvest liquid is concentrated by ultrafiltration, continuous flow centrifugation, column chromatography, cracking, lysing agent removal, and centrifugation. The process of antigenic protein. [0003] At present, most of the purification processes of domestically produced influenza vaccines use a combination of zonal rotor density gradient centrifugation and column chromatography for purification, or only density gradient centrifugation, or column chromatography; Density gradient centrifuges are limited by the capacity of a single batch. In large-scale production, it is usually necessary to use multiple centrifugation operation...

AI Technical Summary

Problems solved by technology

[0003] At present, most of the purification processes of domestically produced influenza vaccines use a combination of zonal rotor density gradient centrifugation and column chromatography for purification, or only density gradient centrifugation, or column chromatography; Density gradient centrifuges are limited by the single capacity. In large-scale production, it is usually necessary to use multiple centrifugation operations in batches. The operation is cumbersome, the workload is heavy, and it is easy to cause the problem of poor uniformity of the same batch of samples.

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