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Mouse-specific anti-fertility polypeptide

An anti-fertility and composition technology, which is applied in the direction of specific peptides, peptides, diseases, etc., to achieve the effect of immune anti-fertility, no toxic and side effects, and optimistic application prospects

Inactive Publication Date: 2014-12-10
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 sequences

[0024] In order to screen out polypeptide fragments that have both mouse-specificity and good antigenicity, the present invention uses Invitrogen's online polypeptide analysis tool (http: / / peptideselect.invitrogen.com / peptide) and the Protean analysis tool in DNAStar analysis software , The protein sequences of human / mouse epididymis protein HE6, sperm protein Sp38 and Slc26a8 were analyzed. According to the principles of selecting polypeptides with good specificity, antigenicity, strong hydrophilicity, and avoiding structures located in the helical region, the amino acid sequences at positions 4-19 and 40-54 of the Sp38 protein, and amino acid sequences at positions 2-20 and 36- The 51st amino acid sequence and the 3-14 and 35-50 amino acid sequences of HE6 protein were studied. The results of the analysis of the polypeptide sequence by the Protean analysis tool in the DNAStar analysis software ...

experiment example 2

[0027] Experimental example 2 Anti-fertility research of polypeptide immunized mouse serum antibody of the present invention

[0028] Reagents and their preparation methods:

[0029] Coating diluent composition and preparation: composed of 0.05mol / L sodium carbonate-sodium bicarbonate buffer solution, 1.5g of sodium carbonate and 2.9g of NaHCO 3 After mixing, add double distilled water to 1000mL and adjust to pH9.6.

[0030] Blocking solution composition and preparation: 5% BSA-PBS solution, BSA5g, add PBS (pH7.4) 100mL to obtain phosphate buffered saline (PBS):

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

[0032] B liquid: 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 was dissolved in double distilled water 1000mL.

[...

experiment example 3

[0053] Experimental example 3 Anti-fertility effect of the 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] Grouping of experimental animals: 50 healthy Balb / c mice (provided by the 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 used in combination, the samples are grouped according to the grouping design Table 2 in Experimental Example 2. The doses of each polypeptide are mixed in equal amounts, and the total dose used each time is 100 μg / only. Group 5 mice were immunized with the same method and dose as follows.

[0058] Immunization procedure: For the first immunization, the polypeptide was mixed with Freund’s complete adjuvant in an equal volume at a dose of 100 μg...

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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 comprising mouse-specific epididymis protein and sperm protein. Background technique: [0002] According to World Health Organization (WHO) statistics at present, there are more than 17 billion rats in the whole world, and more than 4 billion rats are arranged in my country, which has caused serious harm to agriculture, forestry, animal husbandry and human health. According to the report of the Food and Agriculture Organization of the United Nations (FAO), the world's food loss due to rodents can reach up to 15%-20% of the harvest, which is equivalent to the gross national income of 25 poor countries and enough for 150 million people's annual rations. In my country, nearly ten million mu of forest and one billion mu of grassland are destroyed into deserts or barren islands every year, and the loss exceeds 3 billio...

Claims

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Application Information

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Patent Type & Authority Patents(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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