Immunopotentiator and preparation method and application thereof
An immune enhancer and technology for immunizing animals, which can be used in antiviral agents, pharmaceutical formulas, medical preparations containing active ingredients, etc., and can solve problems such as animal disease, poor immunogenicity, and low titer of neutralizing antibodies
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Embodiment 1
[0025] Preparation of Immunopotentiator for Rabies Oral Live Vaccine
[0026] (1) Add 1g of chitosan to 100ml of 0.8% acetic acid solution, mix thoroughly, and use 7.5% NaHCO 3 Adjust the pH value of the solution to 7.0, adjust the pH slowly to prevent precipitation, then set the volume to 100ml, pressurize at 121°C for 20min, and prepare a 1% chitosan solution. Mix the high-pressure chitosan solution with the virus solution 1:1, which is the rabies immune enhancer 1.
[0027] (2) Weigh 50 mg of α-mannan peptide and add it to 10 ml of PBS at pH 7.2, mix well, dissolve fully, and then filter to sterilize at 0.22 μm. Take 0.5 ml of 5 mg / ml α-mannan peptide solution and add it to 2 ml of virus liquid, which is the rabies immune enhancer 2.
[0028] (3) Add 100mg of trimethyl quaternary ammonium chitosan (TMC) to 10ml of PBS with a pH value of 7.2, mix well, and after fully dissolving, pressurize at 121°C for 20min, add 1ml of TMC after high pressure to 1ml of virus liquid Amon...
Embodiment 2
[0032] Toxicity Test of Immunopotentiator to Rabies Virus
Embodiment approach
[0034] 1) The virus titer was 10 8.3 TCID 50 The rabies virus / ml is diluted 10 times with serum-containing DMEM medium, that is, 10 -1 , 10 -2 ,·····,10 -9 .
[0035] 2) Mix the immunopotentiator and the virus solution with the rabies virus solution at a ratio of 1:1, and measure the TCID50 of the mixed solution virus after 1 hour at 37°C in a water bath; at the same time, only the immunopotentiator without adding Virus liquid (immunopotentiator mixed with DMEM medium) and only virus fluid without immunopotentiator as a control.
[0036] 2.TCID 50 The measurement results are shown in Table 1.
[0037] Table 1 Virus fluid TCID50 measurement results
[0038] group lgTCID 50 / ml Immune enhancer 1+ virus solution 7.0 Immune booster 2+ virus solution 7.0 Immune enhancer 3+ virus solution 7.3 Immune enhancer 4+ virus solution 7.3 Immune enhancer 5+ virus solution 7.0 DMEM+virus solution 7.3 Immune booster 5 (without virus) ...
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