A kind of nano vaccine and preparation method thereof
A nano-vaccine and self-assembly technology, applied in nanotechnology, nanotechnology, nanomedicine, etc., can solve the problem of reducing the efficacy of adjuvant/tumor-specific antigen-peptide combination therapy, and cannot guarantee the simultaneous acceptance of immune adjuvant and tumor-specific Antigenic peptides, increased immune-related toxic and side effects, etc.
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Embodiment 1
[0049] A preparation method of nano vaccine, comprising the steps of:
[0050] 1. Synthesis of polypeptide-maleimide-docosahexaenoic acid / peptide-mal-DHA
[0051] 1.1 Prepare the antigen peptide for later use, the N-terminal of the antigen is connected to the -Cys-(a)-(b) sequence, (a) is a hydrophilic group, and (b) is a cathepsin cleavage site group;
[0052] 1.2N-(2-Aminoethyl)maleimide and docosahexaenoic acid undergo a condensation reaction to obtain an intermediate product I;
[0053] 2,2'-Dithiodiethanol and docosahexaenoic acid are dissolved in dichloromethane, the condensing agent is 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, the base is Under the condition of N,N-diisopropylethylamine, the reaction was carried out at 43°C for 24 hours, the solvent was spin-dried, and the intermediate product I (DHA-mal) was obtained after separation and extraction through a silica gel column;
[0054] 1.3 The final product peptide-mal-DHA is obtained by Michael addition reactio...
Embodiment 2
[0067] The preparation method of embodiment 2 nano vaccine
[0068] (A) A linker sequence CSSVVR was added to the front of the sequence of model antigen ovalbumin (ovalbumin, OVA, amino acid sequence SIINFEKL) 257-264, which was synthesized by Zhejiang Ontolais Biotechnology Co., Ltd.;
[0069] N-(2-aminoethyl)maleimide and linolenic acid are subjected to a condensation reaction to obtain an intermediate product I, and then the intermediate product I and the antigen are subjected to a Michael addition reaction to obtain OVA covalently coupled linolenic acid: wherein The cysteine on C is used to connect the highly hydrophobic DHA, so that the modified peptide is in the form of one end hydrophobic and one end hydrophilic, which is conducive to the formation of self-assembled nanoparticles; the VVR sequence is the cleavage site of cathepsin S , when the nano-vaccine particles were endocytized by DC cells, the model antigen OVA was released under the action of cathepsin S in the...
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