Preparation method of new coronavirus S1+ S2 anti-idiotype egg yolk antibody vaccine

An anti-idiotype, yolk antibody technology, applied in the direction of anti-viral immunoglobulin, antibody, anti-viral agent, etc., can solve the problems of weak potency, difficult to decompose, and small molecular weight of the new coronavirus vaccine, and achieve strong potency and stability , the effect of long retention time in the body

Pending Publication Date: 2020-08-25
西安咸辅生物科技有限责任公司
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Problems solved by technology

[0009] The present invention provides a method for preparing a new coronavirus S1+S2 anti-idiotype egg yolk antibody vaccine, which solves the problem that the molecular weight of the new coronavirus vaccine in the prior art is small, it is not easy to be decomposed when it enters the body, the stabi...

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  • Preparation method of new coronavirus S1+ S2 anti-idiotype egg yolk antibody vaccine
  • Preparation method of new coronavirus S1+ S2 anti-idiotype egg yolk antibody vaccine
  • Preparation method of new coronavirus S1+ S2 anti-idiotype egg yolk antibody vaccine

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Embodiment 1

[0030] The embodiment of the present invention provides a preparation method of a novel coronavirus S1+S2 anti-idiotype egg yolk antibody vaccine, please refer to the attached figure 1 , the preparation method comprises:

[0031] Step 110: using the S1+S2 protein of the new coronavirus to prepare anti-S1+S2 antibodies;

[0032] Specifically, the pathogenic mechanism of the new coronavirus is mainly caused by the spike protein, which has two subunits S1 and S2, of which S1 contains a receptor binding domain (RBD) , which is responsible for recognizing membrane receptors (angiotensin-converting enzyme 2, ACE2) on human cells; S2 contains the essential elements required for the membrane fusion process. The new coronavirus combines RBD with ACE2 on human cells to produce membrane fusion, thereby entering human cells and causing human infection. The purpose of the present invention is to prepare the anti-idiotypic yolk antibody of S1+S2 subunits, screen out the β-type anti-idioty...

Embodiment 2

[0106] In order to detect the titer and specificity of the rabbit anti-S1+S2 antibody and the mouse anti-S1+S2 antibody, this example carried out titer detection on the rabbit anti-S1+S2 antibody and the mouse anti-S1+S2 antibody and specific detection.

[0107] 1. Potency detection

[0108] In this example, ELISA method was used to detect the titer of the rabbit anti-S1+S2 antibody and the mouse anti-S1+S2 antibody.

[0109] (1) Use the ELISA method to detect the titer of the above-mentioned rabbit anti-S1+S2 antibody, specifically:

[0110] Step 610: Dilute the S1+S2 protein to 10 μg / mL with coating buffer, and add the diluted S1+S2 protein solution into 8 reaction wells;

[0111] Step 620: After standing at room temperature for 40-60 minutes, discard the liquid in the 8 reaction wells, and wash the 8 reaction wells three times with PBS for 3 minutes each time;

[0112] Step 630: Dilute the rabbit anti-S1+S2 antibodies by 10 1 、10 2 、10 3 、10 4 、10 5 、10 6 、10 7 、1...

Embodiment 3

[0144] In order to detect the titer, specificity and concentration of the refined anti-idiotypic yolk antibody, this embodiment carried out titer detection, specificity and concentration detection on the refined anti-idiotypic yolk antibody.

[0145] 1. Potency detection

[0146] The refined product of the S1+S2 anti-idiotypic yolk antibody in this example is prepared by using the rabbit anti-S1+S2 antibody, that is, the anti-S1+S2 antibody in step 110 is a rabbit anti-S1+S2 antibody. The specific detection steps are:

[0147] Step 810: After diluting the mouse anti-S1+S2 antibody 1000 times with coating buffer, add it to 8 reaction wells, add 100 μL to each reaction well, and let stand at room temperature for 1 hour;

[0148] Step 820: Discard the liquid in the 8 reaction wells, and wash the 8 reaction wells three times with PBS for 3 minutes each time;

[0149] Step 830: Dilute the refined S1+S2 anti-idiotypic yolk antibody by 10 with PBS 1 、10 2 、10 3 、10 4 、10 5 、10...

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Abstract

The invention discloses a preparation method of a new coronavirus S1+ S2 anti-idiotype egg yolk antibody vaccine. The preparation method comprises the following steps: preparing an anti-S1 + S2 antibody by adopting S1+ S2 protein of new coronavirus; diluting the anti-S1 + S2 antibody by using PBS, and mixing the diluted anti-S1 + S2 antibody with a Freund' S incomplete adjuvant in an equal volumeto obtain a first mixed solution; performing intramuscular injection on a laying hen by using the first mixed solution, and performing first immunization on the laying hen; judging whether the layinghen meets a second predetermined condition or not; if the laying hen meets the second predetermined condition, determining the laying hen as a high-immunity hen; collecting high-immunity eggs according to a third predetermined condition; performing disinfection, egg yolk separation and egg yolk tissue crushing on the high-immunity eggs to obtain an egg yolk liquid; removing impurity protein in theegg yolk liquid to obtain an anti-idiotype egg yolk antibody crude product; and purifying the anti-idiotype egg yolk antibody crude product to obtain an anti-idiotype egg yolk antibody refined product. The preparation method achieves the technical effects that the new coronavirus vaccine is large in molecular weight, is not easily decomposed after entering a human body, and is long in-vivo retention time, high in stability and high in efficacy.

Description

technical field [0001] The invention relates to the technical field of epidemic prevention, in particular to a preparation method of a novel coronavirus S1+S2 anti-idiotypic yolk antibody vaccine. Background technique [0002] The novel coronavirus is currently prevalent in almost all countries in the world, and the number of people infected exceeds several million, and it is still increasing so far, causing huge damage to human life safety and social economy. At present, the main way to deal with the epidemic is early diagnosis, early isolation and early treatment. These measures have had a positive effect on preventing the spread of the new crown virus, but they have also had a great negative impact on all walks of life, and countries around the world have paid a huge price for this. [0003] The best way for human beings to prevent the epidemic of this infectious disease and finally eliminate it is to develop an effective vaccine as soon as possible. So far, no new coro...

Claims

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Application Information

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IPC IPC(8): C07K16/10C07K16/02A61K39/42A61P31/14
CPCC07K16/10A61P31/14A61K2039/505C07K2317/11C07K2317/10
Inventor 黄威权田超赵锋李丹黄晓文欧阳森
Owner 西安咸辅生物科技有限责任公司
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