Short chain polypeptide, application thereof as vaccine adjuvant and vaccine with short chain polypeptide serving as adjuvant
A technology of vaccine adjuvant and short peptide, which is applied in the field of short peptide, its application as a vaccine adjuvant, and the field of vaccines using the short peptide as a vaccine adjuvant, which can solve the problem of inability to form hydrogel and vaccine adjuvant to wrap antigen And other issues
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preparation Embodiment 1
[0052] Preparation of vaccine vac-1 loaded with OVA protein in short peptide hydrogel at pH 7.4 and room temperature 20°C
[0053] (1) Synthesis of D-configuration short peptide Fbp-G by FMOC-solid-phase synthesis method D f D f D Y, the structural formula is as follows:
[0054]
[0055] Specific steps are as follows:
[0056] 1) Weigh 0.5mmol 2-cl-Trt resin into a solid phase synthesizer, add 10mL of anhydrous dichloromethane (hereinafter referred to as DCM), place on a shaker and shake for 5min to fully swell the 2-Cl-Trt resin ;
[0057] 2) Remove the DCM from the solid-phase synthesizer equipped with 2-Cl-Trt resin with ear washing ball;
[0058] 3) Dissolve 0.75 mmol of Fmoc-protected amino acid in 10 mL of anhydrous DCM, add 0.75 mmol of DIEPA, then transfer to the above-mentioned solid-phase synthesizer, add 0.75 mmol of DIEPA, and react at room temperature for 1 h;
[0059] 4) Sealing: Remove the reaction solution in the solid-phase synthesizer with ear wash ...
preparation Embodiment 2
[0069] Short peptide hydrogel Car-G at pH 7.4 and room temperature 20°C D f D f D Preparation of vaccine vac-2 loaded with OVA protein
[0070] (1) Synthesis of D-configuration short peptide Car-G by FMOC-solid-phase synthesis method D f D f D Y, the structural formula is as follows:
[0071]
[0072] The specific steps are as described in Preparation Example 1, and the end-capping group is carprofen.
[0073] Its structural characterization data are as follows:
[0074] 1 H NMR (400MHz,DMSO)δ11.32(s,1H),9.19(s,1H),8.56–8.30(m,2H),8.20–7.96(m,3H),7.86(d,J=8.3Hz, 1H), 7.51–7.30(m, 3H), 7.26–7.01(m, 11H), 6.89(t, J=6.3Hz, 2H), 6.65(d, J=8.3Hz, 2H), 4.62–4.35(m ,3H),3.83–3.41(m,4H),2.98(dd,J=9.7,4.0Hz,1H),2.78–2.56(m,3H),2.33(t,J=11.5Hz,1H),1.35( d,J=5.8Hz,3H).
[0075] (2) Take 1mg Car-G D f D f D Put Y in a 1.5 ml glass bottle, add 400 microliters of PBS solution (pH=7.4), adjust the pH value to 7.4 with sodium carbonate solution, heat to boiling to completely ...
Embodiment 1
[0091](1) Mice were immunized for the first time
[0092] Take a mouse of 6-8 weeks, record the time of the first injection as 0 day, take the protein prepared in Preparation Example 1, Preparation Example 2, Comparative Preparation Example 1, Comparative Preparation Example 2 and Comparative Preparation Example 3 Vaccines vac-1, vac-2, OVA, Al-OVA, and vac-3 were dispersed into a viscous solution by vortexing the hydrogel, and then administered subcutaneously at the groin of each mouse at a dose of 100 microliters per mouse. injection.
[0093] (2) Second immunization of mice
[0094] At the time point of the 14th day, the protein vaccines vac-1, vac-2, OVA, Al -OVA and vac-3 were dispersed into a viscous solution by vortexing the hydrogel, and then subcutaneously injected into the groin of each mouse at a dose of 100 microliters per mouse.
[0095] (3) Measurement of antibody titer
[0096] On the 21st day, the mouse serum was taken, and the corresponding antibody titer ...
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