Nano vaccine and preparation method thereof
A technology of nano-vaccine and nano-calcium carbonate, which is applied in the field of nano-vaccine and its preparation, can solve the problems of limiting vaccine efficiency, low endocytosis efficiency, and inability to effectively produce anti-tumor effector T cells, so as to improve vaccine efficiency and promote activation effect
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[0046] The present invention also provides a preparation method of the nano-vaccine described in the above-mentioned technical scheme, comprising the following steps:
[0047] a) mixing the nano-calcium carbonate suspension with the cationic polymer solution and placing it aside for complex treatment, then centrifuging and washing to obtain carrier particles;
[0048] b) dispersing the carrier particles in water to obtain a carrier suspension;
[0049] c) After mixing the carrier suspension with the antigen-adjuvant mixture and standing for compound treatment, centrifuge washing and ultrasonic dispersion to obtain a nano-vaccine dispersion;
[0050] Or mix the carrier suspension with the antigen-adjuvant mixed solution and put it aside for complex treatment, then centrifuge and wash to obtain the nano vaccine powder.
[0051] Among them, the characteristics such as the type, content, particle size and source of nano-calcium carbonate, cationic polymer, antigen, and adjuvant a...
Embodiment 1
[0084] 1.1 Preparation of vaccine
[0085] Ultrasonically disperse nano-calcium carbonate in ultrapure water, the concentration of nano-calcium carbonate is 1mg / mL, and the ultrasonic time is 5min to form nano-calcium carbonate suspension. Polyethyleneimine (25000Da, PEI25K) was dissolved in ultrapure water at a concentration of 5 mg / mL to form a polyethyleneimine solution. Add the polyethylenimine solution into the nano-calcium carbonate suspension, mix equal volumes, vortex for 15 seconds, and then let stand at room temperature for 15 minutes; then centrifuge and wash at 6000r / min for 10 minutes and wash twice to obtain the carrier particles.
[0086] The carrier was dispersed in ultrapure water at a concentration of 1 mg / mL. After that, add an equal volume of antigen-adjuvant mixture, the antigen is a commercially available model antigen (ovalbumin OVA antigen), the adjuvant is oligonucleotide CpG, and the mass ratio of carrier:antigen:adjuvant is 1:0.2 :0.1. After vort...
Embodiment 2
[0113] 1.1 Preparation of vaccine
[0114]Ultrasonically disperse nano-calcium carbonate in ultrapure water, the concentration of nano-calcium carbonate is 1mg / mL, and the ultrasonic time is 5min to form nano-calcium carbonate suspension. Polylysine (15000Da) was dissolved in ultrapure water at a concentration of 5 mg / mL to form a polylysine solution. Add the polylysine solution to the nano-calcium carbonate suspension, mix equal volumes, vortex for 15 seconds, and then let stand at room temperature for 15 minutes; then centrifuge and wash at 6000r / min for 10 minutes and wash twice to obtain the carrier particles.
[0115] The carrier was dispersed in ultrapure water at a concentration of 1 mg / mL. After that, add an equal volume of antigen-adjuvant mixture, the antigen is a commercially available model antigen (ovalbumin OVA antigen), the adjuvant is oligonucleotide CpG, and the mass ratio of carrier:antigen:adjuvant is 1:0.2 :0.1. After vortexing for 15 s, let stand at ro...
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