Method for refining chicken egg yolk antibodies against Escherichia coli

A chicken egg yolk antibody and Escherichia coli technology, which is applied in the field of chicken egg yolk antibody purification, can solve problems such as growth retardation, death, and affecting the quality of livestock products

Inactive Publication Date: 2011-01-19
ZHENGZHOU HOUYI PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0002] Piglet Escherichia coli diarrhea is a common infectious disease caused by porcine pathogenic E. On the level of passive treatment of a large number of drugs for the symptoms shown after the onset, the bacteria will develop drug resistance. to hidden danger

Method used

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  • Method for refining chicken egg yolk antibodies against Escherichia coli
  • Method for refining chicken egg yolk antibodies against Escherichia coli

Examples

Experimental program
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Embodiment

[0021] In this example, Escherichia coli K88, K99, and 987P strains were taken as examples to prepare chicken egg yolk antibodies against Escherichia coli K88, K99, and 987P strains. The specific steps are as follows:

[0022] 1) Preparation of pili antigen:

[0023] Take Escherichia coli K88, K99, and 987P strains, culture them in TSB liquid medium to logarithmic phase, centrifuge at 5000r / min at 4°C, discard the supernatant, wash the precipitate with normal saline, repeat centrifugation and washing twice, and collect the bacteria , add formaldehyde according to 0.45% of the cell volume, inactivate at 37°C for 24 hours, add thimerosal containing 0.01% of the cell volume of formaldehyde; add white oil adjuvant according to the ratio of cell: oil adjuvant 1:1, grind into a uniform water-in-oil antigen solution;

[0024] 2) Preparation of anti-Escherichia coli K88, K99, 987P chicken egg yolk antibodies:

[0025] A Immunization: Choose healthy egg-laying non-immune laying hens...

experiment example

[0029] The anti-Escherichia coli chicken yolk antibody obtained in the embodiment of the present invention was used to conduct experiments to detect its various properties and parameters.

[0030] 1. Determination of flora

[0031] Determination of molecular weight: SDS-PAGE electrophoresis, the concentration of the separating gel is 7.5%, the concentration of the stacking gel is 5%, the current is 11mA, the voltage is 200V, and the time is 3.5h. The marker uses a low relative molecular mass standard protein, and the gel is removed for conventional Coomassie brightening. Blue staining, such as figure 1 As shown, the result appears a clear electrophoresis band at 180KD, and its molecular weight is about 180KD.

[0032] Purity analysis: two-way immunodiffusion method, a precipitation line appears in the result (the color difference between the precipitation line and the background of the test result is small, it is difficult to take a photo and generally can only be observed wi...

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Abstract

The invention discloses a method for refining chicken egg yolk antibodies against Escherichia coli, which comprises the following steps of: culturing by adopting formaldehyde, thiomersalate and a white oil adjuvant to prepare antigen solution; immunizing healthy chickens by using the antigen solution, and collecting immune eggs; extracting egg yolk antibodies from the immune eggs; and purifying by adopting ammonium sulfate to obtain the chicken egg yolk antibodies against the Escherichia coli. For the chicken egg yolk antibodies against the Escherichia coli, pili are collected first, and then Escherichia coli antibodies in the chicken egg yolk are collected after organism immunization of chickens, wherein when the Escherichia coli antibodies in the chicken egg yolk are collected, ammonium sulfate solution is adopted for purification. Experiments prove that the potency of the chicken egg yolk antibodies against the Escherichia coli is obviously improved.

Description

technical field [0001] The invention relates to a method for refining chicken egg yolk antibody against Escherichia coli. Background technique [0002] Piglet Escherichia coli diarrhea is a common infectious disease caused by porcine pathogenic E. On the level of passive treatment of a large number of drugs for the symptoms shown after the onset, the bacteria will develop drug resistance. to hidden dangers. Contents of the invention [0003] The purpose of the present invention is to provide a method for purifying chicken egg yolk antibody with higher titer against Escherichia coli. [0004] In order to achieve the above object, the technical solution adopted in the present invention is: a method for refining chicken egg yolk antibody against Escherichia coli, the steps are as follows: [0005] 1) Preparation of pili antigen: [0006] Take the Escherichia coli strain, culture it to the logarithmic phase in TSB liquid medium, centrifuge, discard the supernatant, wash th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/12C07K16/02C07K1/36C07K1/34C07K1/30
Inventor 吴红云李建正韩改会
Owner ZHENGZHOU HOUYI PHARMA
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