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 immaturity, weak immune response, and danger of foreign aid DNA integration, and achieve the effect of improving immunogenicity, skeletal muscle quality and strength.

Inactive Publication Date: 2010-02-03
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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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 gene sequence of the myostatin C-terminal region (133 amino acids) by using the method of synthesizing genes, and constructs two therapeutic vaccines for myostatin, a gene vaccine and a protein vaccine. In order to break the immune tolerance of the 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-end of the myostatin protein vaccine and gene vaccine when constructing the vaccine. 830-844 To enhance the immune effect of the two therapeutic vaccines of myostatin. Taking advantage of the effect of antigen expression vector on antigen immunogenicity, the immunogenicity of gene peptidin was selected when constructing genetic vaccine. Immunization of healthy adult BALB / c mice can induce specific vaccine-specific plasmid pVAC1-cms for human myostatin molecules. The plasmid pQE30 fused to express 6His is us...

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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 constructpQE-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, protein vaccine and gene vaccine construction, animal immunity experiment, in vitro detection of induced antiserum, etc., 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 ontogeny. The study of myogenesis mainly includes the formation and regulation mechanism of muscle tissue, the growth and control factors of muscle tissue, etc. The important significance of myogenesis research is that 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 there are objective morphological and molecular markers to distinguish between proliferation and differentiation. Skeletal muscle cells,...

Claims

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Application Information

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Patent Type & Authority Patents(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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