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Kidney bean agglutinin egg yolk antibody and preparation method and application thereof

A technology of kidney bean lectin and egg yolk antibody, which is applied in the field of kidney bean lectin egg yolk antibody and its preparation, can solve the problems of kidney bean nutrition loss, poor patient sensitivity, non-specificity, etc., and achieve low preparation cost, low cost, and safety high effect

Inactive Publication Date: 2020-03-17
JIANGHAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the general method for the detoxification of kidney bean poisoning is food poisoning, that is, vomiting, gastric lavage and intravenous rehydration, which is non-specific and the patient's sensitivity is poor.
At present, the method to avoid food poisoning of kidney beans is mainly in the cooking method. Heating in boiling water for more than five minutes completely destroys the lectin, and then cooking is the gold standard for kidney bean processing, but long-term heating will lose many nutrients in kidney beans.
At present, the method to avoid poisoning of kidney bean lectin is high-temperature heating method, which brings a lot of trouble to cooking kidney beans, and at the same time greatly reduces the nutritional value of kidney beans
There are often poisoning events caused by eating kidney beans because the lectin is not destroyed. There is no efficient and specific antidote for kidney beans on the market. The detoxification method is a general method for food poisoning
Low efficiency, causing certain discomfort to the body

Method used

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  • Kidney bean agglutinin egg yolk antibody and preparation method and application thereof
  • Kidney bean agglutinin egg yolk antibody and preparation method and application thereof
  • Kidney bean agglutinin egg yolk antibody and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] 1. Preparation of egg yolk antibody

[0029] Take phase bean lectin standard 100ug, emulsify it with an equal volume of Freund's adjuvant, subcutaneously inject laying hens on the back, use Freund's complete adjuvant for the first injection, and boost immunization with Freund's incomplete adjuvant for the 2nd and 3rd injections, Eggs were collected 25-30 days after the first immunogen injection.

[0030] Take the lyophilized soybean lectin, dilute it to an appropriate concentration with sterilized physiological saline, add an equal volume of Freund's complete adjuvant FCA or Freund's incomplete adjuvant FICA, and then use the double push method of the syringe to mix the adjuvant and immune antigen, make it fully mixed into a water-in-oil emulsion,

[0031] 2. Crude extraction of egg yolk antibody

[0032] Remove the egg white from the egg, dilute the egg yolk 9 times by weight with distilled water, adjust the pH to 5.4 with octanoic acid, centrifuge at 9000r / min for 2...

Embodiment 2

[0037] In this example, except that the crude extraction steps of yolk antibody are different, the others are the same as in Example 1, specifically:

[0038] One, the preparation of egg yolk antibody, with embodiment 1

[0039] 2. Crude extraction of egg yolk antibody

[0040] Remove the egg white from the egg, dilute the egg yolk 9 times by weight with distilled water, adjust the pH to 5.2 with caprylic acid, centrifuge at 8000r / min for 20min overnight at 4°C, precipitate the supernatant with 52% ammonium sulfate solution at 4°C overnight, centrifuge at 8000r / min for 30min, remove the supernatant , again with 37% ammonium sulfate for the second precipitation, overnight at 4°C, 8000r / min, centrifuged for 30min, and after removing the supernatant, the white precipitate was crudely extracted IgY.

[0041] 3. The refined preparation of egg yolk antibody is the same as in Example 1. The yolk antibody with a purity of 94% was obtained.

Embodiment 3

[0043] In this example, except that the crude extraction steps of yolk antibody are different, the others are the same as in Example 1, specifically:

[0044] One, the preparation of egg yolk antibody, with embodiment 1

[0045] 2. Crude extraction of egg yolk antibody

[0046] Remove the egg white from the egg, dilute the egg yolk 9 times by weight with distilled water, adjust the pH to 5.6 with octanoic acid, centrifuge at 10,000r / min for 20min at 4°C overnight, precipitate the supernatant with 48% ammonium sulfate solution at 4°C overnight, centrifuge at 10,000r / min for 30min, remove the supernatant , again with 33% ammonium sulfate for the second precipitation, overnight at 4°C, 10000r / min, centrifuged for 30min, after removing the supernatant, the white precipitate was crudely extracted IgY.

[0047] 3. The refined preparation of egg yolk antibody is the same as in Example 1. The yolk antibody with a purity of 93% was obtained.

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Abstract

The invention discloses a kidney bean agglutinin egg yolk antibody and a preparation method and application thereof. The method comprises the steps of (1) treating kidney bean agglutinin, taking the treated kidney bean agglutinin as an immunogen, immunizing an egg laying hen for a period of time, and collecting eggs; (2) removing egg white to obtain yolk, adjusting the pH value to 5.2 to 5.6 by using caprylic acid after diluting, carrying out primary precipitation, centrifuging and taking supernate, adding an ammonium sulfate solution of 48% to 52% for secondary precipitation, centrifuging toremove supernate, adding ammonium sulfate of 33% to 40% for tertiary precipitation, centrifuging, and removing supernate to obtain white precipitate, namely a crude kidney bean agglutinin egg yolk antibody; and (3) purification of the kidney bean agglutinin egg yolk antibody: subjecting the obtained crude product of the kidney bean agglutinin egg yolk antibody to G100 molecular sieve and ion exchange column chromatography, collecting an eluent, and carrying out dialysis and concentration so as to obtain the kidney bean agglutinin egg yolk antibody. The kidney bean agglutinin egg yolk antibodycan effectively recognize and combine kidney bean agglutinin, and shows an excellent detoxification effect on kidney bean agglutinin in animal experiments.

Description

technical field [0001] The invention relates to the technical field of antibody preparation, in particular to a kidney bean agglutinin egg yolk antibody and its preparation method and application. Background technique [0002] The main cause of kidney bean poisoning is that the kidney bean lectin is toxic to the human body. Symptoms of poisoning appear about 30 to 60 minutes after eating, mainly nausea, vomiting and dizziness. Now the general method for detoxification of kidney bean poisoning is food poisoning, that is, vomiting, gastric lavage and intravenous rehydration, which are non-specific and poor in sensitivity to patients. At present, the method to avoid food poisoning of kidney beans is mainly in the cooking method. Heating in boiling water for more than five minutes completely destroys the lectin, and then cooking is the current gold standard for kidney bean processing, but long-term heating will lose many nutrients in kidney beans. The current method to avoid th...

Claims

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Application Information

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IPC IPC(8): C07K16/16C07K16/02C07K1/36C07K1/30C07K1/18C07K1/16A61P39/02
CPCA61P39/02C07K16/02C07K16/16C07K2317/11
Inventor 韩雪陈泽才陈禅友徐勋陈高
Owner JIANGHAN UNIVERSITY
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