Immunocontraceptive synthetic peptide and immunocontraceptive chimeric peptide and application thereof
A technology of immune contraception and chimeric peptide, applied in the field of immunology, can solve problems such as autoimmune diseases, endocrine balance and sexual dysfunction, poor birth control effect, etc., and achieve a strong anti-fertility effect
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Embodiment 1
[0062] Effects of ESP protein and cross-reactive antigen of Ureaplasma urealyticum on fertility
[0063] 1. Materials
[0064] 1.1 Experimental animals and specimens
[0065] 6-month-old (weight about 2.5kg) female New Zealand white rabbits and BALB / c mice (8-12W) were purchased from the Department of Animal Science, Shanghai Jiaotong University School of Medicine, from the Experimental Animal Center of the Chinese Academy of Sciences, and normal human semen samples were obtained from Shanghai Jiaotong University Medicine Provided by the human sperm bank clinic of Renji Hospital Affiliated to the hospital.
[0066] 1.2 Main reagents
[0067] Oligonucleotide sequences and peptides were synthesized by Shanghai Sangon Biotechnology Company, and keyhole limpet hemocyanin (KLH) was purchased from Shanghai Sangon Biotechnology Company. Plasmid pTSA18 was purchased from Shanghai Institute of Family Planning Science and Technology.
[0068] 2.2.1.3 Experimental instruments Mini Pr...
Embodiment 2
[0139] Identification of cross-reactive antigens between UU474 protein of UU and human sperm protein IZUMO
[0140] 1. Experimental animals and materials:
[0141] Sexually mature male and female BALB / c mice (provided by Shanghai Experimental Animal Center, Chinese Academy of Sciences) with reproductive history were used for both in vivo and in vitro reproductive experiments in mice.
[0142] Male New Zealand white rabbits aged 6 months (about 2.5 kg in weight) were purchased from the Animal Science Department of Shanghai Jiao Tong University School of Medicine.
[0143] 2. Main reagents:
[0144] UU liquid medium and solid medium (Shanghai Tongji Hospital Ministry of Railways Key Laboratory of STD); restriction enzymes BamH I, Hind III, EcoR I, T4 DNA ligase, gel recovery DNA kit, reverse transcription kit, La Taq enzyme (Dalian Bao Biological Company); pre-stained low molecular weight standard protein Marker, low molecular weight standard protein Marker (Fermentas); Agoros...
Embodiment 3
[0242] Study on the function of chimeric peptide immunocontraceptive vaccine
[0243] 1. Materials Synthesis Peptide CP1 was synthesized by Shanghai Sangon Biotechnology Company.
[0244] Complete Freund's adjuvant and incomplete Freund's adjuvant, horseradish peroxidase (HRP)-labeled goat anti-rabbit IgG, FITC-labeled goat anti-rabbit IgG, Anti-His monoclonal antibody (Sigma company) PVDF membrane ( Millipore), ECL bioluminescent chromogen (Millipore).
[0245] 6-month-old (weight about 2.5kg) female New Zealand white rabbits and mature BALB / c mice (8-12W) were purchased from the Experimental Animal Center of the Chinese Academy of Sciences from the Department of Animal Science, Shanghai Jiaotong University School of Medicine.
[0246] Second, the method
[0247] 1. Design and synthesis of chimeric peptides
[0248] The cross-reactive antigens in the sperm protein Izumo, tNASP, and ESP are ATLTK, IERLT, and TGGFT, respectively, and the three short peptides hESP 103-112 (TGG...
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