A method of obtaining a concentrate of microbial cells in the production of plague vaccine

 

The invention relates to the field of immunology. Native culture of Y. pestis grown by the method of deep cultivation. Then take away the sediment and put it microfiltration. The microfiltration is performed with pore size 0.2 μm in the tangential mode of fluid flow at an operating pressure of 0.15-0.2 MPa and productivity of its filtrate 6-8 DM3h-1. The method allows to speed up the process of obtaining a concentrate of microbial cells and simultaneously to increase the output of concentrate per unit volume of medium. 3 table.

The invention relates to the production technology medical immuno-biological preparations, and in particular to methods of obtaining a concentrate of microbial cells in the production of plague live dry vaccine.

A known method of obtaining a concentrate of microbial cells of Y. pestis by growing the microbial mass on the solid nutrient media and flushing its environment drying (Regulation of production 37-86 "plague Vaccine live dry"). In common with the claimed method is to obtain a concentrate of microbial cells, suitable for the preparation of plague vaccine.

The disadvantages of this method include the need for flushing microbial masucci metabolism, what affects the quality of vaccines.

Closest to the claimed is a method of obtaining a concentrate of microbial cells in vaccine production plague live dry (Regulation of production 377-97 "plague Vaccine live dry"), providing for the cultivation of the native culture of the plague microbe, primary sedimentation of microbial biomass in the apparatus of the fermenter for 6 hours at a temperature of 18...20oWith the selection of sediment in the apparatus precipitator or 20-liter carboys and concentration deposition within 18...48 hours at a temperature of 4...8oC. the Yield of the concentrate is 2... 3% of the volume of the medium. In common with the claimed method is to obtain a concentrate of microbial cells, suitable for the preparation of plague vaccine.

The disadvantages of this method include its duration and relatively low yield of concentrate per unit volume of medium.

The objective of the invention is to accelerate the process of obtaining a concentrate of microbial cells with a simultaneous increase in output of concentrate per unit volume of medium.

This object is achieved in that the concentration of microbial mass is carried out by filtration of the culture of the plague microbe through membranes with pore size 0 the ski solutions and prototype shows what is common to them is the process of cultivation of microbial cells in a liquid nutrient medium, and the initial deposition of biomass directly in the apparatus of the fermenter.

The difference of the proposed method is that the process of obtaining a concentrate of microbial cells in the production of plague vaccine is carried out by filtration of the culture of the plague microbe in the tangential mode of fluid flow through a membrane with pore size of 0.2 μm.

The possibility of using microfiltration concentration of microbial cells in the production of plague vaccines installed by us experimentally and unknown from available sources of information.

The essence of the proposed method consists in the following. Native culture vaccine strain of the plague microbe after completion of the cultivation process are left in the machine-fermenter for primary sedimentation at 6 o'clock Deposition takes place at a temperature of 18...20oC. Then the precipitate is collected in 20-liter carboys and subjected to filtration on membranes with pore size of 0.2 μm at a temperature of 18...20oWith a working pressure of 0.15. ..0.2 MPa, the performance of the filtrate 6...8 DM3h-1.

Characteristicsare characteristics of sediment microbial cells of the prototype method for SPM 377-97.

The results, presented in table.1, it is seen that concentrate the microbial cells obtained by the method of microfiltration, due to its characteristics is not inferior cooked in the traditional way and meets the requirements documentation.

The causal connection between the set of essential features of the claimed object and achievable technical result is presented in table.2.

The invention allows to obtain a concentrate of microbial cells due to their physical-chemical and biological characteristics suitable for use in the manufacture of plague vaccine.

Enablement of the claimed invention is shown in the following examples.

Example 1. Obtaining a concentrate of microbial cells.

Native culture of Y. pestis strain EV, grown by the method of deep cultivation in the apparatus of the fermenter V=0,250 m3for 27 hours at a temperature of 26...28oS, has the following characteristics: pH=7,4 pH, the concentration of microbial cells (TOC) turbidity gisk named after. L. A. Lytvyn) 40 billion cells ml-1, TFM is missing. After completion of the culturing microbial slurry in the apparatus of the fermenter for 6 hours at a temperature of 18...20oC. For the traditional system Sartocon-mini" through membranes with pore size of 0.2 μm at a temperature of 18...20oWith a working pressure of 0.15. . . 0.2 MPa, the performance of the filtrate 6...8 DM3h-1. In the filtering process, after reducing the amount of microbial sediment three times, every 10 minutes take sterile samples of 5 ml to determine the concentration of microbes in the TOC turbidity gisk named after. L. A. Lytvyn. When reaching a concentration of 150. . .170 billion CL ml-1the microfiltration process is stopped. The total duration of the concentration of two 20-liter bottle is 12. . .18 o'clock Exit concentrate microbial cells 8...10 l (3.5...4% of the volume of the medium).

Example 2. The concentration of the selected semi-finished plague vaccine (concentration of microbial cells).

Cultivation of microbes Y. pestis is carried out as indicated in example 1, however, due to various reasons is unable to grow native culture with the one required by the regulations of the concentration of microbial cells (TOC) turbidity gisk named after. L. A. Lytvyn), which leads to the rejection of a given culture. The use of microfiltration method allows to concentrate the native culture with a low content of microbial cells to microbial suspensions with the required concentration. Primary sedimentation and filtration are carried out as described in example 1. Lifespan is .5 liters (1.5 to 2% of the volume of the medium).

Example 3. Concentration plaguicides (sedimentation-resistant crops.

Cultivation of microbes Y. pestis is carried out as described in example 1. Native culture in its characteristics in accordance with the regulations, but when the deposition apparatus is precipitators or 20-liter bottles after 48 h the precipitate is formed, which does not allow microbial suspension with the required concentration of microbes. The entire volume newsademic culture is subjected to microfiltration as described in example 1 to obtain a microbial suspension with the desired characteristics. The output of concentrate microbial cells is 8...10 l (3.5...4% of the volume of the medium).

Example 4. Characteristic of the plague vaccine live dry, prepared with the use of the proposed method of obtaining a concentrate of microbial cells and the prototype method.

Characteristic of the plague vaccine live dry, prepared with the use of the proposed method of obtaining a concentrate of microbial cells and the prototype method, is given in tab.3.

Claims

A method of obtaining a concentrate of microbial cells in the production of plague vaccine, involving the deposition of biomass in the apparatus of the fermenter and the end of the pores of 0.2 μm in the tangential mode of fluid flow at an operating pressure of 0.15-0.2 MPa and performance filtrate 6-8 DM3h-1.

 

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