Preparation method of ACE2 anti-idiotypic egg yolk antibody vaccine
A yolk antibody and anti-idiotypic technology, applied in the direction of antibodies, anti-enzyme immunoglobulin, chemical instruments and methods, etc., can solve problems such as weak efficacy, toxic side effects, and retention time in the body
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[0027] A preparation method of ACE2 anti-idiotypic yolk antibody vaccine, the preparation method comprising: uniformly mixing equal volumes of ACE2 protein solution and Freund's incomplete adjuvant to obtain a first mixed solution; using the first mixed solution to prepare anti- ACE2 antibody; after diluting 0.2mL of the anti-ACE2 antibody into 5mL with PBS buffer, mix it with Freund's incomplete adjuvant in equal volume to obtain a second mixed solution with the first concentration; use the second mixed solution Intramuscularly inject laying hens, carry out the first immunization of the first predetermined number of times to the laying hens according to the first predetermined condition; judge whether the laying hens are high-free chickens; if they are high-free chickens, carry out the last time On the third day after the first immunization, high-free eggs were collected for 20 consecutive days; wherein, the high-free eggs were eggs produced by the high-free chickens; the high...
Embodiment 1
[0034] The embodiment of the present invention provides a preparation method of ACE2 anti-idiotypic yolk antibody vaccine, please refer to the attached figure 1 , the preparation method includes steps S100-S1000, specifically:
[0035] Step S100: Mix the ACE2 protein solution and Freund's incomplete adjuvant in equal volumes to obtain the first mixed solution;
[0036] Step S200: using the first mixed solution to prepare anti-ACE2 antibody;
[0037] Specifically, the ACE2 protein is human ACE2 protein, purchased from Beijing Sino Biological Technology Co., Ltd. The incomplete Freund's adjuvant was used to mix with the ACE2 protein solution in equal volumes, and the first mixed solution was obtained after thorough stirring. The first mixed solution was used to prepare anti-ACE2 antibodies, and there were two kinds of anti-ACE2 antibodies, namely rabbit anti-ACE2 antibodies and mouse anti-ACE2 antibodies. Wherein the anti-ACE2 antibody prepared from mice is a mouse anti-ACE2 ...
Embodiment 2
[0116] In order to detect the titer and specificity of the anti-ACE antibody, this embodiment carried out the titer detection and specificity detection of the anti-ACE antibody.
[0117] 1. Potency detection
[0118] In this embodiment, the titer detection of the rabbit anti-ACE2 antibody and the mouse anti-ACE2 antibody is carried out by ELISA method.
[0119] (1) ELISA is used to detect the titer of the rabbit anti-ACE2 antibody, including steps T110-T190, specifically:
[0120] Step T110: Dilute the ACE2 protein to 10 μg / mL with coating buffer, add the diluted ACE2 protein solution into 8 reaction wells, and let stand at room temperature for 1 hour;
[0121] Step T120: Discard the protein solution in the 8 reaction wells, wash the 8 reaction wells three times with PBS, and wash for 3 minutes each time;
[0122] Step T130: Dilute the rabbit anti-ACE2 antibody by 10 1 、10 2 、10 3 、10 4 、10 5 、10 6 , 10 7 、10 8 times, and then added to 8 of the reaction wells respect...
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