Method for preparing tetanus toxoid vaccine

Abstract

The invention discloses a method for preparing a tetanus toxoid vaccine. According to the process, with clostridium tetani strains as raw materials, the tetanus toxoid vaccine is prepared through the following steps of: culturing of tetanus toxoid, bacterium liquid separation, ultrafiltration and concentration, salting out, ultrafiltration desalting and the like. According to the method, firstly culture liquid is subjected to virus-free treatment and then refined, so that the porous channel plugging caused by accumulation of thalli and other impurity segments at a plate and frame membrane package during plate and frame filtering to remove thalli is reduced, and the smoothness during filtration is increased; toxoid protein and other allergens in the culture liquid are removed by changing the salting-out method; and as the desalting methods of the culture liquid after concentration and salting out adopt the tangential flow ultrafiltration method, the destruction of antigen caused by shearing of toxoid protein is reduced, and the protein precipitation is avoided. By utilizing the method, the time for preparing the tetanus toxoid vaccine is shortened, and the production efficiency is improved.

Description

technical field [0001] The invention relates to a preparation method of a tetanus toxoid vaccine. Background technique [0002] Tetanus toxoid is a protein produced by Clostridium tetani and secreted outside the bacteria. It consists of 1315 amino acids and has a molecular weight of 150-700Da. The original tetanus toxoid was originally used for human immunization. This toxoid has a good immune effect, but the side effects after vaccination are very large, and there are even cases of death from anaphylactic shock. This is mainly due to the existence of a large number of incompletely hydrolyzed protein components in the original toxoid, which cause allergic reactions. In order to reduce the side effects of vaccination, Ramon refined and purified the original tetanus toxoid in 1923, and prepared a refined tetanus toxoid. Observations by use showed that the side effects of the refined tetanus toxoid were significantly less than those of the original toxoid after vaccination. ...

AI Technical Summary

Problems solved by technology

[0003] Refined tetanus toxoid needs to be detoxified during production. Since the original toxoid contains a large amount of medium components, it is easy to cross-link the toxin molecule through formaldehyde during the detoxification process, making purification more difficult. In addition, the detoxified Large volume, inconvenient for refining operation

Centrifuge separation or filtration is usually used in the refining process, but the resulting vaccine products have poor selectivity, low quality, low purity (400-600 flocculent units per mg of protein nitrogen), dark color, and can be checked by polyacrylamide electrophoresis , still a mixture of components

[0004] At present, in the production process of tetanus toxoid vaccine in domestic enterprises, the processes of separation of bacterial liquid, concentration of tetanus toxoid solution, purification of tetanus toxoid, and desalination are relatively simple and extensive. Some processes do not meet the requirements of GMP, and some processes are in Use under mass production conditions will be limited

For example, in the bacterial liquid separation process, the volume to be processed in general large-scale production must reach more than 500L. If centrifugation is used, it is not suitable, the processing time will be very long, and a lot of labor is consumed; if canvas filtration is used for sterilization, the filtration is not complete. The defect that the canvas material does not meet the GMP requirements; the disadvantage of plate and frame filtration is that the filter plate is easy to block and the filtration time is too long; larger

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