Method for protecting viral vaccine and improving thermal stability utilizing heavy water

A virus vaccine, thermal stability technology, applied in the direction of virus/bacteriophage, biochemical equipment and methods, microorganisms, etc., can solve the problems of uncertain stabilizing effect, different suitable concentrations, and suitable concentrations to be determined, etc., to improve thermal stability. , the effect of improving the quality of vaccines and improving the stability of the virus

Inactive Publication Date: 2005-12-07
ZHEJIANG PUKANG BIOTECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For heavy water, it can improve the stability of many relatively temperature-labile viruses, but for viruses with relatively temperature-resistant biological characteristics such as hepatitis A virus, the stabilization effect is uncertain
In addition, the appropriate concentration of heavy water for different DNA viruses and RNA viruses is also different, and the appropriate concentration for H2 attenuated strains of hepatitis A has yet to be determined.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] by D 2 O treatment of deuterated-HAV-H2 strain to the confirmation of high temperature (70 ℃, 80 ℃) stability improvement: 1, preparation D 2 O-KMB 17 Cell: KMB 17 Cells were seeded in an area of ​​38 cm 2 In a small square bottle, culture with ordinary growth medium, grow into a single layer after 3 days at 37 ° C, discard the original culture medium, and replace it with 36% D 2 O maintenance solution, maintained at 36°C for 48 hours. Among them, the common growth solution uses LH as the mother solution, the final concentration of calf serum is 10%, the final concentration of eagle’s is 5%, and the pH is 7.1; D in the maintenance solution 2 O concentration 36%, calf serum 2%, eagle's content 5%, pH 7.5.

[0025] 2. Prepare the deuterated hepatitis A-H2 attenuated strain, discard the original maintenance solution of the cells, add 1ml of virus seed diluted 1:20, absorb at 37°C for 2 hours, add 36% D 2 The maintenance solution of O was cultured at 35°C for 28 days,...

Embodiment 2

[0032] by D 2 O-treated deuterated-HAV-H 2 Confirmation of improved stability of the strain at 37°C:

[0033] 1. Preparation D 2 O-KMB 17 Cell: KMB 17 Cells were seeded in an area of ​​37 cm 2 In a small square bottle, culture with ordinary growth medium, grow into a single layer after 3 days at 37 ° C, discard the original culture medium, and replace it with 36% D 2 O maintenance solution, maintained at 36°C for 48 hours. Among them, the common growth solution uses LH as the mother solution, the final concentration of calf serum is 10%, the final concentration of eagle’s is 5%, and the pH is 7.1; maintenance solution D 2 O concentration 36%, calf serum 2%, eagle's content 5%, pH 7.5.

[0034] 2. Preparation D 2 Olated HAV-H 2 Attenuated strain: Discard the original cell maintenance solution, add 1ml of virus seed diluted 1:20, absorb at 37°C for 2 hours, add 36% D 2 O maintenance solution, cultured at 35°C for 28 days, during which the maintenance solution was chang...

Embodiment 3

[0049] 1. The same batch number (batch number 20000909) of several live attenuated hepatitis A vaccines was mixed and left at 40,000rpm for 4 hours, then the original MEM solution was poured out, and an equivalent amount of 85% D 2 O Placed at 8-10°C for 5 months, sampling CCID every month 50 / ml value, the control group uses H 2 O, the results are shown in the table below:

[0050] Table 3 Vaccines with 85% D 2 The titer (CCID) after O was placed as a suspension at 8-10°C for 1-5 (months) 50 / ml) value

[0051] Time (months) D H Titer drop D H

[0052] 0 6.17 6.38 / /

[0053] 1 6.17 6.17 0.00 0.21

[0054] 2 5.83 5.63 0.34 0.75

[0055] 3 5.63 5.17 0.54 1.21

[0056] 4 5.50 4.83 0.67 1.55

[0057] 5 5.50 4.63 0.67 1.75

[0058] (t test, t=-1.31, 0.5>P>0.2)

[0059] It can be seen from Table 3 that with time, the titer drop in group D was significantly smaller than that in group H, indicating that the vaccine stability in group D was higher than that in the control ...

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PUM

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Abstract

Disclosed is a method for protecting viral vaccine and improving thermal stability utilizing heavy water, which consists of choosing the concentration of heavy water (D2O) by a predetermined proportion, processing vaccines at different stages during vaccine production, deuterating the cells with heavy water, thus obtaining deuterated virus vaccine, or suspending the virus vaccine directly with high concentration heavy water (D2O).

Description

(1) Technical field [0001] The invention is a method for protecting virus vaccines with heavy water and improving the stability of heat-resistant virus vaccines, especially for protecting live attenuated hepatitis A vaccines and enhancing their heat resistance. (2) Background technology [0002] The instability of live virus vaccines has been troubling people since humans discovered that virus vaccines can effectively prevent viral diseases. Live vaccines must maintain potency (infectivity) during storage and transportation so that they can replicate in the body after vaccination, so they need to be stored in a cold chain. The cost of the cold chain includes not only a large amount of refrigeration equipment, but also the cost of maintaining the system. According to statistics, this cost accounts for about 8% of the vaccination cost of the recipients, which is very important for the developing third world. is a burden. Therefore, there is an urgent n...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/02
Inventor 陈悦青毛江森庄昉成唐彩华柴少爱钱汶
Owner ZHEJIANG PUKANG BIOTECH
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