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Hepatitis b vaccine and preparation technology thereof

A hepatitis B and vaccine technology, applied to medical preparations containing active ingredients, viral peptides, cells modified by introducing foreign genetic material, etc., can solve the problem of preS antigen not being very immunogenic

Active Publication Date: 2015-01-28
DOBEEL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the resulting linear soluble form of the preS antigen is not very immunogenic
Therefore, attempts to improve immunogenicity were not successful compared to complete surface antigens in which the preS antigen and the S antigen are co-expressed in particle form and the preS antigen is located on the outer surface of the particle

Method used

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  • Hepatitis b vaccine and preparation technology thereof
  • Hepatitis b vaccine and preparation technology thereof
  • Hepatitis b vaccine and preparation technology thereof

Examples

Experimental program
Comparison scheme
Effect test

example

[0085] Hereinafter, the present invention will be described in more detail with examples. However, these examples are for illustrative purposes only, and the present invention is not intended to be limited to these examples.

example 1

[0086] The preparation of example 1 hepatitis B vaccine

[0087] 1.1 Preparation of recombinant complete surface antigen (preS antigen and S antigen; L-HBsAg)

[0088] (1)-1 clone

[0089] Polymerase chain reaction (PCR) was used as a template to amplify the envelope gene (preS1-preS2-S) and a poly The complete 3'-UTR coding region (SEQ ID NO. 1) of the polyadenylation site was then introduced into an expression vector. At this time, polymerase chain reaction was performed by using a Pfu DNA polymerase, and primers were prepared to amplify the coding region of HBsAg and the complete 3'-UTR [forward primer: 5-GGA AGA TCT CAA TCT CGG GAA-3 (SEQ ID NO.2), reverse primer: 5-GGA AGA TCT CGA ATA GAA GGAAAG-3 (SEQ ID NO.3), the recognition sequence of BglII is underlined]. A polymerase chain reaction product of about 2.75 kbp was obtained, which was coordinated to a pMSG vector linearized by BglII enzyme (see Korean Patent Application No. 10-2000-0043996 and PCT / KR01 / 01285). ...

example 2

[0105] Comparison of Example 2 Recombinant Complete Surface Antigen (L-HBsAg) Immunogenicity

[0106] To determine whether the recombinant whole surface antigen (L-HBsAg) of the present invention is highly immunogenic and induces a strong immune response, animal experiments were conducted to compare with known second-generation hepatitis B vaccines.

[0107] 2.1 Comparison of immunogenicity

[0108] (1) Purpose: To compare the antibody titer and immune response induced by the recombinant L-HBsAg antigen.

[0109] (2) Materials and methods

[0110] (a) Experimental animals: 6-week-old female C57BL / 6 mice.

[0111] (b) Experimental vaccines

[0112] The entire surface antigen (L-HBsAg) prepared and purified according to Example 1-1 was adsorbed on alum to prepare a test vaccine. As a control group, a recombinant S antigen (Hepavax-Gene, Green Cross Co.) produced by Hansenula polymorpha, and a recombinant S antigen (recombinant B-type B) produced in CHO cells without preS ant...

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Abstract

The invention discloses an expression vector expressing a recombinant complete HBV surface antigen comprising L protein, M protein and S protein, prepared by injecting a polynucleotide of a coding region which has a complete HBV envelope gene and a complete 3'-UTR nucleotide containing polyadenylation sites into a pSGM vector identified by the preservation number KCCM 10202.

Description

[0001] divisional application [0002] This application is based on PCT application number PCT / KR2008 / 000518, international filing date 2008.01.28, priority date 2007.01.31, date of entering the national phase 2009.07.20, application number 200880002629.2, and the title of the invention is "a hepatitis B vaccine and The divisional application filed by its preparation process". technical field [0003] The present invention relates to an anti-hepatitis B virus (HBV) hepatitis B vaccine, which includes complete HBV surface antigen, the surface antigen is composed of preS antigen and S antigen, a composite HBV antigen vaccine, the composite HBV antigen vaccine Besides the complete surface antigen, it further includes an HBV core antigen, and a preparation method thereof. Background technique [0004] HBV infection leads to acute and chronic hepatitis. Most HBV-infected patients can fully recover from viral infection after one to two months, but about 10% of HBV-infected pati...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/85C12N5/10C12P21/02C07K14/02A61K39/29A61P31/20
Inventor 廉晶善安秉喆赵显镇金桐姸李周娟金基贤尹在承文洪模
Owner DOBEEL CO LTD
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