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Method for efficiently culturing and expressing hepatitis B surface antigen based on recombinant hansenula polymorpha cell technology

A technology of hepatitis B surface antigen and Hansenula, which is applied in the field of medical bioengineering to achieve the effect of high antigen expression, high expression and high cell fermentation density

Inactive Publication Date: 2018-10-30
DALIAN HISSEN BIO-PHARM CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Table 1 The disadvantages of hepatitis B vaccine cells constructed by existing gene recombination technology

Method used

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  • Method for efficiently culturing and expressing hepatitis B surface antigen based on recombinant hansenula polymorpha cell technology
  • Method for efficiently culturing and expressing hepatitis B surface antigen based on recombinant hansenula polymorpha cell technology
  • Method for efficiently culturing and expressing hepatitis B surface antigen based on recombinant hansenula polymorpha cell technology

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Embodiment 1

[0053] A method for efficiently cultivating and expressing hepatitis B surface antigen based on recombinant Hansenula cell technology, said method comprising the steps of:

[0054] Primary seed cultivation steps:

[0055] Inoculate 1mL of the recombinant Hansenula working seed batch engineering strain into 350mL of the primary medium, culture at 30°C for 20h, and the shaking table rotates at 200rpm to obtain the primary seed culture;

[0056] Secondary seed cultivation steps:

[0057] Inoculate 200mL of primary seed culture into 3L of secondary seed culture medium, culture at 30°C for 20h, and shaker at 200rpm to obtain secondary seed culture;

[0058] Three-level seed cultivation steps:

[0059] Take 3L of secondary seed culture and inoculate it into 30L of tertiary seed medium, culture at 30°C for 13 hours, increase the stirring speed from 200rpm to 400rpm, the pH value is 5.05, the air flow rate is 10L / min, and the dissolved oxygen is not less than 20%. obtaining a terti...

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Abstract

The invention relates to a method for efficiently culturing and expressing hepatitis B surface antigen based on a recombinant hansenula polymorpha cell technology, and belongs to the technical field of medical bioengineering. The method includes the steps of primary seed culture, secondary seed culture, tertiary seed culture and fermentation culture. The cell density of a recombinant hepatitis B vaccine (hansenula polymorpha) after cell expansion and culture expression is higher than that of an existing gene recombinant hepatitis B vaccine after cell expansion and culture expression, HBsAg expression quantity is higher, HBsAg granule structures are complete, and cell granules are uniform in size.

Description

technical field [0001] The invention relates to a method for efficiently cultivating and expressing hepatitis B surface antigen based on recombinant Hansenula cell technology, which belongs to the technical field of medical bioengineering. Background technique [0002] Viral hepatitis type B (viral hepatitis type B, referred to as hepatitis B) is caused by hepatitis B virus (HBV), with fatigue, loss of appetite, nausea, vomiting, oily, hepatomegaly and abnormal liver function as the main clinical manifestations. Some cases have fever and jaundice; a few cases have a protracted course of disease and become chronic, or develop into liver cirrhosis or even liver cancer; in severe cases, the disease progresses rapidly and may develop into severe hepatitis; other infected people become asymptomatic virus carriers. [0003] Hepatitis B virus is called hepatitis B virus for short. It is a DNA virus belonging to the hepadnaviridae family. According to current knowledge, HBV is onl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/19C12N15/81C12P21/02C07K14/02C12R1/78
CPCC07K14/005C12N15/815C12N2730/10122C12P21/02
Inventor 李贵兴张平陈新亭李纯厚姜晓娟刘立斌
Owner DALIAN HISSEN BIO-PHARM CO LTD
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