A kind of vaccine adjuvant and its application
An adjuvant and antigen technology, applied in the field of veterinary biological products, can solve the problems of poor protein stability, easy degradation or denaturation, and affect the accuracy of test results, etc., and achieve the advantages of simple operation, good immune response level, and reduced stress response Effect
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[0024] As an embodiment of the present invention, the cellulose ether includes carboxymethyl cellulose, sodium carboxymethyl cellulose, potassium carboxymethyl cellulose, calcium carboxymethyl cellulose, hydroxyethyl cellulose, hypromellose base cellulose.
[0025] As an embodiment of the present invention, in the vaccine adjuvant of the present invention, the acrylic polymer is 2-10 mg / ml in terms of carbomer, and the cellulose ether is 0.1-8 mg / ml.
[0026] As a preferred embodiment of the present invention, in the vaccine adjuvant of the present invention, the acrylic polymer is 4-10 mg / ml in terms of carbomer, and the cellulose ether is 4-8 mg / ml.
[0027] The term "immunopotentiator" (immunopotentiator) means that it is not immunogenic itself, but can be used to stimulate the immune system to improve the response to other immunogenic substances. When used together with the antigen of interest or administered separately to the same subject, it can directly or indirectly m...
Embodiment 1
[0062] Preparation and identification of embodiment 1 antigen
[0063] 1.1 Preparation and content determination of porcine circovirus antigen
[0064] 1.1.1 Preparation of SH strain of porcine circovirus type 2
[0065] Porcine circovirus type 2 SH strain was diluted 1:9 V / V with MEM liquid medium (made with MEM dry powder medium purchased from Invitrogen, USA according to the instructions), and then inoculated in 5% of the volume of the cell culture medium. PK15 (ATCC, preservation number is CCL-33) monolayer cells, adsorb at 37°C for 30 minutes, add cell maintenance solution (add 4% calf serum and 2mmol / L D-glucosamine hydrochloride in MEM liquid medium), 37 Cultivate for 4 days at ℃, freeze and thaw 2-3 times, harvest the virus, and the virus titer is 10 6.5 TCID 50 / ml.
[0066] Cultivate the virus liquid with the spinner bottle cell culture method, with the PK15 cells covered with a single layer, pour off the cell culture liquid (in the MEM liquid medium, add 6% calf...
Embodiment 2
[0094] The preparation of embodiment 2 adjuvants
[0095] Preparation of carbomer mother liquor: dissolve carbomer 971P in distilled water, let it swell overnight at room temperature to obtain 20 mg / ml carbomer solution, add 3M sodium hydroxide solution to adjust the pH value of the solution to 7.2, add appropriate amount of distilled water to adjust Carbomer solution to obtain a carbomer mother solution with a final concentration of 15 mg / ml.
[0096] Preparation of sodium carboxymethylcellulose mother liquor: mix 1g sodium carboxymethylcellulose with 100ml distilled water, heat and boil until completely dissolved, then cool to room temperature, that is, configure sodium carboxymethylcellulose mother liquor with a concentration of 10mg / ml .
[0097] Preparation of ISCOM matrix mother solution: Take 12mg / ml lipid mixture stock solution and add it to PBS solution, then add 30mg / ml QuilA stock solution to make the final concentration of QuilA 15mg / ml, phosphatidylcholine 3mg / ml...
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