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
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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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