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Method for removing residual DNA in hydrophobia vaccine product by utilizing ultrasound combined with EDTA solution

A rabies vaccine and ultrasound technology, applied in the field of removing host DNA and impurity proteins, can solve the problems of large side effects of vaccines, low rate of removing impurity proteins, and high content of host DNA, and achieve the goal of removing content, improving product quality and pass rate, and making breakthroughs. The effect of quality bottlenecks

Active Publication Date: 2010-07-21
LIAONING YISHENG BIOLOGY PHARMACY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] 1. The residual host DNA content after virus vaccine extraction is too high
[0005] 2. Viral vaccines still contain too much host protein after extraction, and the removal rate of miscellaneous proteins is not high, resulting in relatively large side effects after clinical use of the vaccine
[0006] 3. The above defects have affected the quality of virus vaccine products and restricted the production of vaccine products
[0009] The result is only a slight effect, without substantially solving the problem

Method used

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  • Method for removing residual DNA in hydrophobia vaccine product by utilizing ultrasound combined with EDTA solution
  • Method for removing residual DNA in hydrophobia vaccine product by utilizing ultrasound combined with EDTA solution
  • Method for removing residual DNA in hydrophobia vaccine product by utilizing ultrasound combined with EDTA solution

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Experimental program
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Effect test

Embodiment 1

[0024] Select 40ml of concentrated rabies vaccine concentrated 25 times by ultrafiltration, add 4.444ml of 100mmol / L EDTA solution with a pipette gun to make the final concentration of EDTA 10mmol / L, shake well and let stand at room temperature for 40min, then use Φ6 The Scientz-IID ultrasonic cell pulverizer with the horn performs ultrasonic treatment on the rabies vaccine concentrate, and conducts ultrasound under the protection of an ice bath to ensure that the temperature of the concentrate does not exceed 26°C, and the ultrasonic power is set to 600W (64%). 2s, interval time 3s, total working time 20min (240 times), the batch number of the concentrated solution after ultrasonic is S-20100205. Then it was purified by XK50 chromatographic column and Sepharose 4FF gel chromatography. The detection range of the ultraviolet detection system was set to 0.2A, the output was set to 100mV, and the paper speed of the recorder was set to 2cm / h. Using 10mmol / LPBS with pH 7.6 as the m...

Embodiment 2

[0026] Select 40ml of concentrated rabies vaccine concentrated 25 times by ultrafiltration, add 4.444ml of 10mmol / L EDTA solution with a pipette gun, so that the final concentration of EDTA is 1mmol / L, shake well and let stand at room temperature for 40min, then use Φ6 The Scientz-IID ultrasonic cell pulverizer with the horn performs ultrasonic treatment on the rabies vaccine concentrate, and conducts ultrasound under the protection of an ice bath to ensure that the temperature of the concentrate does not exceed 26°C, and the ultrasonic power is set to 600W (64%). 2s, the interval time is 3s, the total working time is 20min (240 times), the batch number of the obtained ultrasonic concentrate is S-20100207. Then it was purified by XK50 chromatographic column and Sepharose 4FF gel chromatography. The detection range of the ultraviolet detection system was set to 0.2A, the output was set to 100mV, and the paper speed of the recorder was set to 2cm / h. Using 1 mmol / LPBS of pH 7.6 a...

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Abstract

The invention provides a method for removing residual DNA in a hydrophobia vaccine product by utilizing ultrasound combined with EDTA solution. The method solves the problems that a method for extracting hydrophobia vaccine through concentration, purification and the like can only remove dissociative DNA in a certain percentage and cannot remove host DNA combined with antigen protein and clinical adverse reactions commonly occur. The method has main points of: adding the EDTA solution into hydrophobia vaccine concentrate; performing ultrasonic treatment on the hydrophobia vaccine by using ultrasonic wave to make the host DNA broken more easily under the action of the ultrasound; and removing the host DNA through chromatography and purification. The method has the advantages that: on the premise of ensuring the valence of the vaccine, the quality of a vaccine product is improved, mass hybrid protein and the host DNA are removed so that the residual DNA content of the vaccine product is less than 100 pg / dose, and the quality problem in the hydrophobia vaccine industry at present is solved.

Description

technical field [0001] The invention relates to a method for removing host DNA and miscellaneous proteins in vaccine products, specifically a method for removing residual DNA in rabies vaccine products by using ultrasound combined with EDTA solution. EDTA solution is a kind of complexing agent, chemical name is ethylenediaminetetraacetic acid or disodium ethylenediaminetetraacetic acid, EDTA mentioned in the present invention can be ethylenediaminetetraacetic acid, also can be ethylenediaminetetraacetic acid disodium Sodium, or a mixture of both. Background technique [0002] Rabies vaccine is to inoculate rabies virus on suitable animal cells, and release the virus into the culture medium after reproduction, during which some residual host cells or their fragments will also be mixed in the culture medium. After clarification, filtration, concentration and purification After extraction, in the stock solution of rabies virus vaccine, free host DNA and host DNA combined with ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/205A61P31/14
Inventor 徐德启张译程晓耕汪春义苏文全王凯
Owner LIAONING YISHENG BIOLOGY PHARMACY
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