Compound adjuvant system and method for preparing adjuvant
An adjuvant and aluminum phosphate adjuvant technology, applied in the field of medicine, can solve the problems of antigen degradation and lack of feasibility, and achieve good adsorption and desorption capabilities, wide application fields, and excellent vaccine immune effects
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Embodiment 1
[0032] Example 1 Preparation of a new adjuvant combination of aluminum phosphate and CpG with high adsorption rate
[0033] 1. Obtaining Aluminum Phosphate Adjuvants with Different Isoelectric Points
[0034] The reaction solution 1 (a mixed solution of aluminum chloride and sodium dihydrogen phosphate) and the reaction solution 2 (sodium hydroxide solution) were pumped into a glass reactor with a volume of 600 mL by a peristaltic pump respectively. Regulate PO 4 3- / Al 3+ The proportion of components and the added volume of NaOH, the specific information is shown in Table 1 below. The reactor was placed on a magnetic stirrer and stirred at a constant speed. Collect the suspension, let it stand, discard the supernatant, and centrifuge at 8000rpm for 15min to obtain a precipitate. Disperse the precipitate in a certain volume of pure water to obtain an aluminum phosphate adjuvant solution in a certain concentration range.
[0035] Table 1 Summary of reaction information of...
Embodiment 2
[0046] Example 2 Application of aluminum phosphate (AP) and CpG adjuvant in gE immunization vaccine
[0047] Adsorption of aluminum phosphate (AP) adjuvants with PI of 7.7, 8.5, and 9.0 on CpG and gE antigens: First, the gE antigen stock solution was adsorbed on aluminum adjuvant (AP) to prepare gE antigen / Al (w / w) Adsorbed sample at a ratio of 200 μg / 1200 μg / ml; then to this gE antigen / Al adsorbed sample was added different concentrations of CpG samples: 1) 0 μg / ml, 2) 100 μg / ml, 3) 200 μg / ml, 4) 300 μg / ml ml, 5) 500 μg / ml, 6) 600 μg / ml, 7) 900 μg / ml, 8) 1200 μg / ml, after 24 hours of adsorption, centrifuge to get the supernatant, and calculate CpG and gE by westen-blot and UV second-order derivative respectively the adsorption effect.
[0048] By the experimental results image 3 It can be seen that after the antigen is adsorbed by the aluminum phosphate adjuvant system with PI greater than 7.7, the concentration of free gE antigen in the preparation after adsorption is <10...
Embodiment 3
[0050] The antigen desorption method of embodiment 3
[0051] 1) Screening of desorption liquid types.
[0052] The adsorption sample whose gE / Al (Al element in aluminum adjuvant) (w / w) (AH adjuvant) ratio is 200μg / 1200μg / ml is desorbed under the desorption conditions as shown in Table 3, and the desorption ratio is 1:3. liquid. Desorb the centrifuged supernatant for 2h, 24h, and 48h at 25 degrees Celsius to detect the UV second-order guide gE content, and calculate the recovery rate. The results are as follows Figure 5 shown.
[0053] For the adsorbed sample with a CpG / Al (w / w) (AH adjuvant) ratio of 600 μg / 1200 μg / ml, add the desorption solution at a ratio of 1:3. After desorption at 25 degrees Celsius for 2 hours, 24 hours, and 48 hours of centrifugation, the supernatant was used to detect the content of UV second-order guide CpG, and the recovery rate was calculated. The results are as follows Figure 5 shown.
[0054] pass Figure 5 The data shows that the desorpti...
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