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Recombinant fusion proteins of hepatitis B core proteins and tuberculosis antigen or antigen fragments and application thereof

A technology of fusion protein and composition, applied in the field of genetically engineered recombinant fusion protein, can solve problems such as inability to induce protective immune response

Active Publication Date: 2015-01-07
INST OF BIOENG ACAD OF MILITARY MEDICAL SCI OF THE CHINESE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, some researchers also found (Stukova MA, Sereinig S, Zabolotnyh NV, Ferko B, Kittel C, Romanova J, et al. Vaccine potential of influenza vectors expressing Mycobacterium tuberculosis ESAT-6protein.Tuberculosis (Edinb) 2006; 86(3-4) : 236-46.), the results of ESAT-6 immunization depended on the method of vaccine delivery: when immunized in the form of peptide or DNA vaccine alone, no significant protective immune response could be induced in animals

Method used

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  • Recombinant fusion proteins of hepatitis B core proteins and tuberculosis antigen or antigen fragments and application thereof
  • Recombinant fusion proteins of hepatitis B core proteins and tuberculosis antigen or antigen fragments and application thereof
  • Recombinant fusion proteins of hepatitis B core proteins and tuberculosis antigen or antigen fragments and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0047] Example 1: Taking the full-length tuberculosis antigen ESAT-6 inserted between amino acids 78-79 of the hepatitis B core protein molecule as an example, a recombinant fusion protein of hepatitis B core protein and tuberculosis antigen carrying a tuberculosis antigen was prepared.

[0048] 1. Construction of HE6 gene

[0049] The tuberculosis antigen ESAT-6 has a total of 94 amino acids, and the corresponding nucleic acid sequence is 282bp in length. With the help of the carrier protein in the plasmid pET21a-H144L2, the Msc I enzyme in the nucleic acid sequence corresponding to the 68-70 amino acids of the hepatitis B core protein (HBc) Site and overlap extension PCR technology, by dividing ESAT-6 into two segments, first amplify the upstream sequence E segment with a length of about 329bp and the downstream sequence H segment with a length of about 190bp, and then through the complementation of the two partial sequences , the entire ESAT-6 gene fusion was introduced, an...

Embodiment 2

[0066] Example 2: Taking HE6 as an example to carry out immunological evaluation of recombinant fusion protein on animal model.

[0067] 1. Animal experiments

[0068] Thirty 4-5-week-old sterile female BALB / c mice were randomly divided into 5 groups, and the immune vectors HBc-N144, HE6, and rESAT-6 were injected subcutaneously, and the immune dose of each protein was 20μg antigen / 200μl / mouse, the first three immunizations were separated by two weeks, and the mice were sacrificed 10 days after the fourth immunization, and the spleen was taken to carry out the detection of cellular immunity. Since the immune carrier HBc is also reported that it may have the effect of immune adjuvant, a group of HE6 without adjuvant control is also set in this embodiment, and HBc-N144 or blank group (blank) are all used as negative controls. Both groups used aluminum hydroxide adjuvant (the final concentration of aluminum was 1.0 mg / ml). The serum of the mice after three immunizations was ta...

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Abstract

The invention relates to one or more types of recombinant fusion proteins formed by hepatitis B core proteins and mycobacterium tuberculosis antigens or antigen fragments. The tuberculosis antigens or antigen fragments are inserted in the same or different permission sites of hepatitis B core proteins in a mode of single antigen or antigen fragment or various types of antigens or antigen fragments which are combined together. The recombinant fusion proteins may comprise a plurality of cofactors. In addition, the invention also relates to the type of the fusion of the tuberculosis antigen or antigen fragments and cofactors and hepatitis B core proteins, nucleic acid and amino acid sequences of coded corresponding recombinant fusion proteins, a method for preparing the recombinant fusion proteins and application of the recombinant fusion proteins in the preparation of medicaments and vaccines for preventing and / or treating tuberculosis.

Description

technical field [0001] The invention relates to a series of gene engineering recombinant fusion proteins, in particular to the recombinant fusion proteins for preventing and / or treating tuberculosis. [0002] In the recombinant fusion protein involved in the present invention, the function of the hepatitis B core protein is to provide a protein carrier that allows the combination of various tuberculosis antigens or antigen fragments to enhance the immune response against Mycobacterium tuberculosis infection. [0003] The invention provides the nucleotide sequence and amino acid sequence encoding the series of recombinant fusion proteins, as well as the preparation method of the corresponding recombinant fusion proteins. [0004] The invention also relates to the application of the series of recombinant fusion proteins in the preparation of vaccines and medicines for preventing and / or treating tuberculosis. Background technique [0005] Tuberculosis is a zoonotic chronic inf...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/10A61K39/04A61K39/39A61K47/48A61P31/06A61K47/42
Inventor 陈薇徐俊杰殷瑛张军董大勇李浩侯利华付玲
Owner INST OF BIOENG ACAD OF MILITARY MEDICAL SCI OF THE CHINESE
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