Composition for preventing and controlling pseudomonas aeruginosa infection and preparation method thereof
A technology of Pseudomonas aeruginosa and composition, which is applied in the field of composition for preventing and treating Pseudomonas aeruginosa infection and its preparation field, can solve problems such as being difficult to put into practical use, achieve good nutritional prospects, improve immunogenicity and protection, and prevent other diseases Effect of infection
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Embodiment 1
[0064] Example 1: Preparation of Pseudomonas aeruginosa O-specific polysaccharide
[0065] Lipid A was separated from O-specific polysaccharides by mild acid treatment.
[0066] Dissolve a small amount of purified LPS of Pseudomonas aeruginosa IATS type 1 in 1% acetic acid solution and put it in a boiling water bath for digestion. After the digestion is completed, cool it to room temperature and put it in a refrigerator at 2-8°C overnight, then add CaCl 2 The solution was transferred to a centrifuge cylinder to a final concentration of 0.01M, placed in a high-speed refrigerated centrifuge, and centrifuged at 15,000 rpm for 2.5 hours at 6°C. The centrifuge cylinder was taken out, and the supernatant was chromatographed on the Sepharose CL-4B gel 16 / 100 column of General Electric Company and analyzed (see figure 1 , figure 2 ).
[0067] On this basis, a large amount of purified LPS was removed from lipid A by the same method as before, and after centrifugation, the supernata...
Embodiment 2
[0068] Example 2: Activation derivatization of Pseudomonas aeruginosa O-specific polysaccharide
[0069] The O-specific polysaccharide of Pseudomonas aeruginosa IATS type 6 strain was dissolved in sodium chloride saline, the concentration was controlled at 10-20 mg / ml, and the pH was adjusted to 5.8 with triethylamine (TEA). Add CDAP dissolved in acetonitrile, which is 1 / 2 of the amount of specific polysaccharide, into the polysaccharide solution. After 30s, adjust the pH to 8.1. Add dissolved 0.1M NaHCO 3 Solution ADH solution to a final concentration of 0.4M. Use 0.1M HCl solution to maintain the pH between 8.05 and 8.15 for 2 hours. After the reaction, the solution was put into dialysis cellophane, and dialyzed with purified water above grade at 7°C overnight, and the water was changed once in the middle. Next day go up General Electric Company SephadexG50 gel 26 / 100 chromatographic column further desalting, make mobile phase with water for injection, collect the first ...
Embodiment 3
[0070] Example 3: Preparation of Pseudomonas aeruginosa O-specific polysaccharide combined with TT carrier protein immunogen
[0071] The O-specific polysaccharide derived from Pseudomonas aeruginosa IATS type 1 strain was dissolved in sodium chloride saline, the concentration was controlled at 6-30 mg / ml, and an equal amount of TT (tetanus toxoid protein, purchased from Chengdu Institute of Biological Products) was added. ), and then add EDAC dissolved in normal saline to a final concentration of 0.1M. The pH was adjusted with 0.1M HCl solution to maintain it at 5.20 ± 0.1. The reaction was stopped after 4 hours. The solution was put into dialysis cellophane, dialyzed overnight at 7°C with purified water above grade, and the water was changed once in the middle. The next day, Sepharose 4FF gel 26 / 100 chromatographic column of General Electric Company was used for separation and purification, and 0.1M sodium chloride solution was used as mobile phase, and the first peak colle...
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