Aluminum hydroxide gel-polysaccharide composite immunologic adjuvant and preparation method and application thereof
A compound immune adjuvant and aluminum hydroxide gel technology, which is applied in the field of biomedicine, can solve the problems of incomplete understanding of the mechanism of action of aluminum salt adjuvants, and achieve the effects of strong immune activity, high immune effect, and high clinical safety
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Embodiment 1
[0042] Example 1 Screening and preparation of compound adjuvants
[0043]The present invention compares liposomes, aluminum hydroxide gels, hydrogels, mannans, polysaccharides (derived from Escherichia coli or Bacillus typhi), etc. individually or in combination as adjuvants to add antigens, and finds that aluminum hydroxide gels / Bacterial polysaccharide (Al-PS) combination as a compound adjuvant can effectively induce both humoral immunity and cellular immunity.
[0044] This compound adjuvant uses two components: aluminum hydroxide gel adjuvant, which is a kind of Al-containing adjuvant. 3+ The inorganic salt has good adsorption effect and can adsorb soluble antigens on the surface of aluminum glue molecules; it is also a good precipitant, which can concentrate antigens and reduce the injection dose. Aluminum hydroxide gel is the most widely used adjuvant and the only adjuvant approved by the FDA for human vaccines so far. Polysaccharides, especially bacterial-derived pol...
Embodiment 2
[0060] Example 2 The present invention uses E. coli polysaccharide or typhoid polysaccharide as an animal immunization test for hepatitis B vaccine adjuvant
[0061] 1. Grouping
[0062] 1. Individual protein group: HBsAg protein 0.1 μg / mouse / time;
[0063] 2. Al(OH) 3 Group: HBsAg protein 0.1μg+Al(OH) 3 25μg / mouse / time;
[0064] 3. E.coli polysaccharide group: HBsAg protein 0.1 μg+E.coli polysaccharide 20 μg / mouse / time;
[0065] 4. Al(OH) 3 +E.coli polysaccharide group: HBsAg protein 0.1μg+Al(OH) 3 25μg+E.coli polysaccharide 20μg / mouse / time;
[0066] 5. PBS group: PBS+Al(OH) 3 25μg+E.coli polysaccharide 20μg / mouse / time.
[0067] 6-week-old C57BL / 6 mice (five mice in each group) were immunized according to the above groups, once every two weeks, three times in total. Blood was collected from the tail vein at 0, 1, 3, 5, 7, 9, 13, and 17 weeks after immunization, respectively, and the serum was separated and stored at -20°C for testing.
[0068] 2. Determination of ...
Embodiment 3
[0083] Embodiment 3 The present invention uses E. coli polysaccharide or typhi polysaccharide as bFGF adjuvant animal immunization test
[0084] 1. Grouping
[0085] 1.bFGF+Al(OH) 3 Group: bFGF protein 20μg+Al(OH) 3 500μg / mouse / time;
[0086] 2. bFGF+E.coli polysaccharide group: bFGF protein 20μg+E.coli polysaccharide 20μg / mouse / time;
[0087] 3. bFGF+Al(OH) 3 +E.coli polysaccharide group: bFGF protein 20μg+Al(OH) 3 500μg+E.coli polysaccharide 20μg / mouse / time;
[0088] 4. bFGF + Freund's adjuvant group: bFGF protein 20μg + Freund's adjuvant / mouse / time;
[0089] 5. PBS group: PBS+Al(OH) 3 500μg+E.coli polysaccharide 20μg / mouse / time.
[0090] 6-week-old C57BL / 6 mice (seven mice in each group) were immunized according to the above groups, once every two weeks, three times in total. Blood was collected from the tail vein at 0, 1, 2, 3, 4, 5, and 6 weeks after immunization, and the serum was separated and stored at -20°C for testing. Humoral immunity and cellular immuni...
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