Therapeutic vaccine for Myostatin specific antibody and its preparation method

A therapeutic vaccine and specific technology, applied in the field of medicine and biology, can solve the problems of weak immune response, immaturity, danger of foreign aid DNA integration, etc., and achieve the effect of improving the quality and strength of skeletal muscle and improving immunogenicity.

Inactive Publication Date: 2007-10-24
FOURTH MILITARY MEDICAL UNIVERSITY
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Problems solved by technology

[0060] However, there are still many deficiencies in DNA vaccines: ①The ability to stimulate the body's immune response is weaker than natural infection
②The expression of the target gene is not ideal; ③How long the expression of the protein in the body can last needs to be further studied; ④The foreign aid DNA introduced into the body has the potential risk of integration, and DNA immunization may cause the dysfunction of the immune system itself
[0070] The immune and pathological mechanisms of vaccines are relatively complicated, such as the immune response activated by therapeutic vaccines, whether the cells are protective or pathological is not clear, etc., resulting in the effect of vaccine therapy is not very stable and ideal, and it is not yet clear in practical applications. not mature enough

Method used

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  • Therapeutic vaccine for Myostatin specific antibody and its preparation method
  • Therapeutic vaccine for Myostatin specific antibody and its preparation method
  • Therapeutic vaccine for Myostatin specific antibody and its preparation method

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Embodiment Construction

[0104] The invention artificially synthesizes the myostatin C-terminal region (133 amino acid) gene sequence by using the synthetic gene method, and constructs two kinds of therapeutic vaccines, a gene vaccine and a protein vaccine against the myostatin. In order to break the immune tolerance of myostatin vaccine and enhance the immunogenicity of the vaccine, a universal Th cell epitope TT derived from tetanus toxin and capable of activating Th cells was inserted into the N-terminus of the myostatin protein vaccine and gene vaccine. 830-844, to enhance the immune effect of myostatin two therapeutic vaccines. Utilizing the influence of antigen expression vectors on the immunogenicity of antigens, the immunogenicity of the gene vaccine was selected for the construction of genetic vaccines. By immunizing healthy adult BALB / c mice, the specific vaccine-specific plasmid pVAC1-cms for human myostatin molecules can be induced, and the plasmid pQE30, which is fused to express 6His, is...

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Abstract

The invention discloses a curative vaccine for Myostatin specific antibody and the method for preparing the same. The human myostatin C-end gene section that is connected with Th cell auxiliary epitope TT830-844 at N-end is prepared by using artificial DNA synthesizing method. The genetic sequence is TT-Ms, and the TT-Ms genetic section is cloned to pQE30 pronucleus expression carrier to construct pQE-TT-Ms recombinant expression carrier, and constructs pQE-Ms recombinant expression carrier by PCR augmenting Ms genetic section. The fused protein His-TT-Ms and His-Ms is expressed in bacillus coli, and is used as protein vaccine for myostatin after certification. The synthesized TT-Ms genetic section is cloned to specific plasmid of pVAC1-cms gene vaccine, and constructs pVAC-TT-Ms recombinant plasmid, and it is used as myostatin gene vaccine after external certification for target protein expression. The constructed Myostatin protein vaccine and gene vaccine is used in healthy adult BALB/c mouse, the specific antibody of myostatin molecule for human can be induced, and the quality and force of skeletal muscles of mouse is obviously improved.

Description

technical field [0001] The invention belongs to the field of medical biotechnology, and relates to technologies such as gene synthesis, construction of protein vaccines and gene vaccines, animal immunization experiments, and in vitro detection of induced antiserum, in particular to a therapeutic vaccine of Myostatin specific antibody and a preparation method thereof. Background technique [0002] Myogenesis refers to the formation and growth of muscle tissue during individual development. The study of myogenesis mainly includes the formation and regulation mechanism of muscle tissue, the growth and control factors of muscle tissue, etc. The significance of myogenesis research lies in: myogenesis has always been one of the hot areas of developmental biology research. Skeletal muscle cells can be induced to differentiate under in vitro culture conditions, and at the same time, there are objective morphological and molecular markers to distinguish...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/395A61K48/00C12N15/62C12N15/70C12N15/74A61P21/00
Inventor 张英起唐量韩苇李萌薛晓畅孟洁如包春杰郝强李维娜王增禄
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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