Redox sensitive polypeptide based on cell-penetrating peptide and application of redox sensitive polypeptide in vaccine vector
A vaccine carrier and membrane-penetrating peptide technology, which is applied in the fields of biomedical materials and immunotherapy, can solve the problem of inability to cause cellular immunity, and achieve the effects of being conducive to large-scale production, good reproducibility, and strong cellular immunity
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Embodiment 1
[0033] Embodiment 1: the design of polypeptide:
[0034] In the present invention, the advantages of the cell-penetrating peptide and the redox-responsive disulfide bond cross-linking agent are combined, and a polypeptide vaccine carrier is designed for antigen delivery. The specific structure of the polypeptide is: Cys-Trp-Trp-Arg8-Cys-Arg8-Cys-Arg8-Cys (including 2 tryptophans, 4 cysteines and 24 arginines: Cys-Trp-Trp- Arg-Arg-Arg-Arg-Arg-Arg-Arg-Arg-Cys-Arg-Arg-Arg-Arg-Arg-Arg-Arg-Arg-Cys-Arg-Arg-Arg-Arg-Arg-Arg-Arg- Arg-Cys). The positively charged polypeptide carrier can combine with the negatively charged antigen protein through electrostatic attraction to form a nanocomplex, which is like a cage to wrap the antigen molecule. In nanocomposites, the sulfhydryl groups of cysteine are spontaneously oxidized to form disulfide bonds, and cross-linking occurs between polypeptides to form denser polypeptide / antigen condensates. The overall positively charged polypeptide / a...
Embodiment 2
[0035] Embodiment 2: the proportioning of polypeptide and ovalbumin OVA (purchased from Sigma-Aldrich)
[0036] Table 1 Particle size and potential of peptide / OVA with different mass ratios
[0037] The mass ratio of peptide and OVA
Embodiment 3
[0038] Embodiment 3: the preparation of vaccine preparation
[0039] Using ovalbumin OVA (purchased from Sigma-Aldrich) as the model antigen, vaccines of different formulations were prepared according to the following table, and the vaccine preparations used for subcutaneous injection are shown in Table 2 respectively.
[0040] Table 2 Different vaccine formulations used for subcutaneous injection
[0041]
[0042] Note: The peptide and antigen (ovalbumin OVA) need to be incubated at room temperature for 2 hours before use.
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