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Mouse-specific anti-fertility polypeptide

An anti-fertility and composition technology, applied in the direction of specific peptides, peptides, diseases, etc., to achieve good immune anti-fertility effects, no toxic side effects, and significant antigenic effects

Inactive Publication Date: 2012-12-19
BIOENG RES INST OF THE ACADEMY OF MILITARY MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the cross-reactivity of the immune response triggered by many natural antigens, it cannot well meet the actual clinical needs. Therefore, in order to improve the specificity of protein antigens, it is necessary to make selections at the epitope level to retain and improve specific polypeptide fragments. In order to get more ideal vaccine target molecules

Method used

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  • Mouse-specific anti-fertility polypeptide
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  • Mouse-specific anti-fertility polypeptide

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0023] Experimental Example 1 Screening of anti-fertility polypeptide sequence

[0024] In order to screen out peptide fragments that have both mouse-specific and good antigenicity, the present invention uses Invitrogen's online peptide analysis tool (http: / / peptideselect.invitrogen.com / peptide) and the Protean analysis tool in DNAStar analysis software , To analyze the protein sequences of human / mouse epididymal protein HE6, sperm protein Sp38 and Slc26a8. According to the principle of selecting good specificity, antigenicity, strong hydrophilicity, and avoiding peptides located in the helical region, the final selection of the 4-19 and 40-54 amino acid sequences of Sp38 protein, and the 2-20 and 36- positions of Slc26a8 protein The 51 amino acid sequence and the 3-14 and 35-50 amino acid sequences of HE6 protein are studied. The results of the analysis of the peptide sequence by the Protean analysis tool in the DNAStar analysis software are shown in figure 1 , The sequence resu...

experiment example 2

[0027] Experimental Example 2 Anti-fertility study of serum antibodies in mice immunized with polypeptides of the present invention

[0028] Reagents and preparation methods:

[0029] Coating diluent composition and preparation: It is composed of 0.05mol / L sodium carbonate-sodium bicarbonate buffer, 1.5g sodium carbonate and 2.9g NaHCO 3 After mixing, add double distilled water to 1000 mL and adjust to pH 9.6.

[0030] The composition and preparation of the blocking solution: 5% BSA-PBS solution, 5g of BSA, add 100mL of PBS (pH7.4) to obtain phosphate buffered saline (PBS):

[0031] A solution: 0.2mol / L sodium dihydrogen phosphate aqueous solution, the preparation method is NaH 2 PO 4 , H 2 27.6 g of O was dissolved in 1000 mL of double distilled water.

[0032] B solution: 0.2mol / L disodium hydrogen phosphate aqueous solution, the preparation method is Na 2 HPO 4 ·7H 2 O 53.6g, Na 2 HPO 4 ·12H 2 O71.6g or Na 2 HPO 4 ·2H 2 O 35.6g is dissolved in 1000mL of double distilled water.

[0033...

experiment example 3

[0053] Experimental example 3 Anti-fertility effect of polypeptide immunized mice of the present invention

[0054] Main reagents:

[0055] Freund's complete adjuvant and Freund's incomplete adjuvant (Sigma, USA).

[0056] experimental method:

[0057] Experimental animal grouping: 50 healthy Balb / c mice (provided by Experimental Animal Center of Beijing Academy of Military Medical Sciences), 25 females and 25 males, 6-8 weeks old, 10-16g, randomly divided into 10 groups, SEQ ID 1- When the polypeptides described in 6 are mixed and used, the samples are grouped according to the grouping design table 2 in Experimental Example 2. The dosage of each polypeptide is mixed in the same amount, and the total dosage for each use is 100μg / group separately. A group of 5 mice was immunized with the same method and dose as follows.

[0058] Immunization procedure: For the first immunization, mix the peptide with the same volume of Freund’s complete adjuvant at a dose of 100 μg / mouse / time, fully em...

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Abstract

The invention discloses six immune anti-fertility polypeptide segments, all of which have excellent immunogenicity. The six immune anti-fertility polypeptide segments can stimulate the organisms to generate specific antibodies and induce the anti-fertility function. According to the fertility comparison of antigen polypeptide immunized mice and the immunized mice in a control group (KLH (keyhole limpet homocyanin) control group), immunization by the six polypeptide antigens mixed group can significantly reduce the farrowing rate of the male and the female mice, wherein litter sizes in the male and the female mice is respectively reduced by 62.8% and 50%. The athletic ability and activity of sperms are significantly reduced in the HE6 polypeptide group or the mixed group immunized male mice. The polypeptide with high-efficiency anti-fertility effect establishes the foundation for research of the mouse-specific immune anti-fertility vaccines, and can be used for application of various anti-fertility preparations.

Description

Technical field: [0001] The present invention relates to a group of mouse-specific anti-fertility polypeptides, in particular to immune anti-fertility polypeptides containing mouse-specific epididymal protein and sperm protein. Background technique: [0002] According to statistics from the World Health Organization (WHO), there are more than 17 billion rats in the world, and more than 4 billion rats in our country, which have caused serious harm to agriculture, forestry, animal husbandry and human health. According to the United Nations Food and Agriculture Organization (FAO) report, the world’s food loss caused by rodents can reach up to 15%-20% of the harvest, which is equivalent to the total national income of 25 poor countries and enough food for 150 million people throughout the year. In our country, nearly 10 million acres of forests and 1 billion acres of grasslands are destroyed into deserts or barren continents every year, with losses exceeding 3 billion yuan. In addit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00A61K38/10A61K47/42A61P15/16A61P15/18
Inventor 王玉民汪莉田利源侯小强李秀丽
Owner BIOENG RES INST OF THE ACADEMY OF MILITARY MEDICAL SCI
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