Preparation method of rotavirus inactivated vaccine

Abstract

The invention relates to a preparation method of a rotavirus inactivated vaccine, and belongs to the technical field of biological medicines. The invention provides a preparation method of a rotavirus inactivated vaccine, the preparation method comprises the steps of virus release, virus clarification, virus inactivation, host DNA degradation, virus concentration and virus purification, the virus release step adopts a pressure crushing method to crush host cells, the virus clarification step adopts continuous flow centrifugation, and the virus purification step adopts continuous flow centrifugation to purify the host cells. The virus purification step adopts continuous flow sucrose density gradient centrifugation; according to the preparation method, a pressure crushing method is adopted to crush host cells in the virus release step, compared with freeze thawing crushing, the pressure crushing step is simple, continuous sample injection can be achieved, the operation time is greatly shortened, compared with ultrasonic crushing, the virus activity is not prone to being damaged through pressure crushing, and the crushing effect on the host cells is better; in addition, closed-loop sterile operation can be achieved through pressure crushing, the crushing condition is stable, and large-scale industrial production can be better achieved.

Description

technical field [0001] The invention relates to a preparation method of a rotavirus inactivated vaccine, belonging to the technical field of biomedicine. Background technique [0002] Rotavirus (RV) is a double-stranded RNA virus belonging to the family Reoviridae. It is one of the main pathogens that cause diarrhea in infants and young children under the age of five. It infects hundreds of millions of people worldwide every year and causes dozens of deaths. Ten thousand. In my country, about 10 million infants and young children suffer from rotavirus infectious gastroenteritis every year, accounting for 1 / 4 of the number of infants and young children, and it is the most important pathogen causing severe diarrhea in infants and young children. [0003] A safe and effective vaccine is an important means to control rotavirus infection. Currently, inactivated rotavirus vaccines are commonly used to control rotavirus infection. However, there have been many incidents in histo...

AI Technical Summary

Problems solved by technology

[0005] First, the virus release step often uses repeated freeze-thaw crushing or ultrasonic crushing. Among them, the repeated freeze-thaw crushing method takes a long time to freeze and thaw, and the operation cycle is long; the heat generated by the ultrasonic crushing method is large, which is easy to destroy the virus activity. Moreover, the area of ​​the ultrasonic probe needs to be proportional to the processing capacity, and it is difficult to realize the equal ratio amplification process in the mass production process;

[0006] Second, the virus purification step mainly uses anion exchange chromatography. This process requires high concentration of the virus clarified liquid. The shear force during the concentration process will damage the virus structure and reduce the recovery rate. Moreover, the chromatographic effect of this process is affected by impurities, Affected by factors such as pressure and salt concentration, the separation effect of the chromatographic column will decrease rapidly with the increase of the number of uses. Sub-packing does not guarantee that the chromatographic column is completely consistent. In addition, the chromatographic fluid contains high concentrations of salt ions, and dialysis bags or desalting columns are required for desalination. In the large-scale production process, it will increase costs and reduce recovery.

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