Aluminum hydroxide gel-polysaccharide composite immunologic adjuvant and preparation method and application thereof
A technology of aluminum hydroxide gel and compound immune adjuvant, applied in the field of biomedicine, can solve the problem that the action mechanism of aluminum salt adjuvant is not completely clear, and achieve the effects of strong immune activity, high anti-tumor effect, and high immune effect.
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Embodiment 1
[0042] Example 1 Screening and preparation of composite adjuvants
[0043]The present invention compares liposome, aluminum hydroxide gel, hydrogel, mannan, polysaccharide (E. / bacterial polysaccharide (Al-PS) combination as a compound adjuvant can not only effectively induce the body to produce humoral immunity, but also effectively induce cellular immunity.
[0044] This composite adjuvant uses two components: among them, aluminum hydroxide gel adjuvant is a kind of adjuvant containing Al 3+ The inorganic salt has good adsorption effect, and can adsorb soluble antigens on the molecular surface of aluminum gel; it is also a good precipitant, which can concentrate antigens and reduce the injection dose. Aluminum hydroxide gel is cheap, easy to use, and non-toxic. It is the most widely used adjuvant, and it is also the only adjuvant approved by the FDA for human vaccines. Polysaccharides, especially polysaccharides derived from bacteria, as a natural source of adjuvants, have...
Embodiment 2
[0060] Embodiment 2 The present invention uses E.coli polysaccharide or typhoid polysaccharide as animal immunity test of 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] The 6-week-old C57BL / 6 mice (five mice in each group) were immunized according to the above grouping, once every two weeks, and three times in total. Blood was collected from the tail vein at 0, 1, 3, 5, 7, 9, 13, and 17 weeks after immunization, and the serum was separated and stored at -20°C for testing.
[0068] Second, the determination of p...
Embodiment 3
[0083] Embodiment 3 The present invention uses E.coli polysaccharide or Salmonella typhi polysaccharide as bFGF adjuvant animal immunity 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] The 6-week-old C57BL / 6 mice (seven in each group) were immunized according to the above grouping, once every two weeks, and three times in total. Blood was collected from the tail vein at 0, 1, 2, 3, 4, 5, and 6 weeks after immunization, the serum was separated, and stored at -20°C for testing, and the humoral immunity and cell...
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