Chitin-sodium alginate packed pylorus helicobacterium protein microballs and their preparation
A technology of Helicobacter pylori and sodium alginate, which is applied in the direction of pharmaceutical formulations, bacterial antigen components, medical preparations of non-active ingredients, etc., can solve the problems of poor affinity of hydrophilic antigens, etc. The effect of release function
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Embodiment 1
[0041] Example 1 Preparation of chitosan-sodium alginate-coated Helicobacter pylori protein microspheres
[0042] 1. Take Hp strain M13, culture it in a microaerobic environment (temperature: 37°C; humidity > 95%) in brain heart infusion medium for 24 hours, wash 6 times with 0.05M phosphate buffer saline (PBS, pH7.4), 5000r / Bacteria were collected by centrifugation at 4°C for 10 minutes, and inactivated by adding 5% formaldehyde solution for 24 hours. 200W×30s×10 times of sonication, centrifuged at 15,000g at 4°C for 20min to collect the supernatant, and determined the antigen concentration by Lowry method.
[0043] 2. Preparation of HpWCP-CAMS
[0044] At room temperature, mix the Hp whole bacterial protein prepared in Example 1 with 2% sodium alginate solution, and add vegetable oil, which includes soybean oil, rapeseed oil, peanut oil, sesame oil and the like. The ratio of vegetable oil to sodium alginate emulsion is 2:8, emulsified at 8000r / min for 10min and then added...
Embodiment 2
[0045] Example 2 Detection of chitosan-sodium alginate-coated Helicobacter pylori protein microspheres
[0046] The microspheres prepared above were centrifuged and washed 3 times with distilled water, collected, and vacuum freeze-dried for future inspection.
[0047] 1. Detection of HpWCP-CAMS morphology
[0048] 1) Freeze-drying of MS
[0049] The obtained microsphere suspension was slowly poured into a glass plate, and the liquid level was lower than 1 cm. After pre-freezing in a -40°C refrigerator for 12 hours, place it directly in a pre-cooled vacuum dryer to dry, and slowly raise the temperature to sublimate the water quickly. When the gas pressure indicator shows that no gas is generated, take out the freeze-dried product for inspection.
[0050] 2) Morphological observation of MS before and after drying
[0051] Take MS, lyophilized powder and lyophilized powder reconstituted after washing and centrifuging, and smear a small amount on a glass slide, and observe the ...
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