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Method for removing chick embryo host protein in rabies vaccine product

A technology for rabies vaccine and host protein, which is applied in the field of removing chicken embryo host protein in rabies vaccine products, can solve the problems of aggravating protein aggregation and difficulty in ensuring the final product quality, and achieve the reduction of miscellaneous protein content, cleanliness requirements and operational difficulty Low, high stability effect

Pending Publication Date: 2021-07-16
SHENZHEN WEIGUANG BIOLOGICAL PROD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

(2) Host DNA exacerbates protein aggregation
(3) Ultrasound can only remove part of the impurities
Direct ultrasonic treatment of the concentrated solution can remove some impurity proteins, but high protein aggregates still remain, making it difficult to guarantee the final quality of the product

Method used

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  • Method for removing chick embryo host protein in rabies vaccine product
  • Method for removing chick embryo host protein in rabies vaccine product
  • Method for removing chick embryo host protein in rabies vaccine product

Examples

Experimental program
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Effect test

Embodiment 1

[0031] This embodiment provides a method for removing chicken embryo host protein in rabies vaccine products, including:

[0032] The culture supernatant of the harvested virus chicken embryo cells is clarified and filtered sequentially through a filter element of 3.0 μm to 5.0 μm and a filter element of 0.45 μm to 1.2 μm, and then concentrated by a 100kD to 300kD ultrafiltration membrane bag or hollow fiber, and then used Phosphate buffer solution containing 50mmol / L~300mmol / L sodium chloride and 100mmol / L~300mmol / L arginine with a pH of 7.6~8.0 is diafiltered 5~10 times, and added at a ratio of 1:2000~1:6000 β-propiolactone is inactivated and hydrolyzed, then human serum albumin is added (the final concentration of human serum albumin is 0.75% to 1.5%), and after standing at 2°C to 8°C for 12h to 24h, transfer to In the ultrasonic special bottle (the transferred liquid volume is 1 / 4~2 / 3 of the total volume of the ultrasonic special bottle), an ultrasonic cleaning machine (fr...

Embodiment 2

[0034] Get 30L harvested viral chicken embryo cell culture supernatant through 3.0μm filter element and 1.2μm filter element to carry out clarification and filtration successively, and after carrying out concentrated 5 times treatment through 300kD ultrafiltration membrane bag, adopt containing 150mmol / L sodium chloride and 200mmol / L arginine phosphate buffer solution (pH7.8) was diafiltered 5 times, then concentrated to about 15 times the original volume to obtain a 2000mL concentrate; add 500μL of β-propiolactone at a ratio of 1:4000 , and vibrated at 2°C to 8°C for 24h, then transferred to 37°C and continued to shake for 2h to complete the inactivation hydrolysis and obtain the inactivation hydrolyzate 1.

[0035] Take 20mL inactivated hydrolyzate 1 and pass through the chromatography column xk26 / 70, sepharose 4FF gel filtration chromatography, collect the elution peak 1, use phosphate containing 100mmol / L sodium chloride at pH 7.8 as the mobile phase, and elute The flow ra...

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Abstract

The invention relates to a method for removing chick embryo host protein in a rabies vaccine product. The method comprises the following steps of: filtering and concentrating harvested virus chick embryo cell culture supernatant, performing washing and filtering by using a phosphate buffer solution containing sodium chloride and arginine, and adding beta-propiolactone for inactivating and hydrolyzing; and adding human serum albumin, performing standing at low temperature, and performing ultrasonic treatment and purification to obtain the rabies vaccine product purified liquid without the chick embryo host protein. According to the method, the human serum albumin is added, so that the influence of ultrasonic waves on virus particles can be effectively reduced, and the content of impure protein in the product is further reduced. The inactivated hydrolysate is subjected to ultrasonic treatment through an ultrasonic cleaning machine, the cleanliness requirement is low, the operation difficulty is low, and the stability is high.

Description

technical field [0001] The invention relates to the technical field of vaccine product purification, in particular to a method for removing chicken embryo host protein in rabies vaccine products. Background technique [0002] The rabies virus culture harvest often contains a mixed system of various components. In the preparation of rabies virus vaccine using Vero cells as the culture substrate, the culture harvest liquid is usually clarified and concentrated, and then the DNA impurities in the product are removed by means of ultrasound and chromatography. And the technology that existing chicken embryo method prepares rabies vaccine is that chicken embryo cells are cut into pieces, and the chicken embryo primary fibroblasts after digestion and dispersion are used as culture substrate. Harvested cultures often contain cell debris, free host DNA, and chick embryo host proteins. After the harvest liquid is clarified and filtered, the final product is usually obtained by simpl...

Claims

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Application Information

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IPC IPC(8): A61K39/205A61K47/42A61P31/14
CPCA61K39/12A61K47/42A61P31/14A61K2039/5252C12N2760/20134
Inventor 郭土敬王雪云李慧梁富叶才文黄致翔王义民梁琳琳曹静刘永娣
Owner SHENZHEN WEIGUANG BIOLOGICAL PROD
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