Immunoaffinity method

An immunoaffinity and affinity technology, applied in the field of immunoaffinity method, can solve the problems of loss of target substance, turbidity of analysis sample liquid, clogging of affinity column, etc., to enhance the overall sealing, increase the pressure in the column cavity, prevent the The effect of the blockage of the column cavity

Inactive Publication Date: 2015-12-09
MARINE FISHERIES RES INST OF ZHEJIANG
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

When the target exceeds the antigen limit that the antibody in the column can bind, the target will be lost, and the concentration of the target cannot be analyzed effectively
[0006] 3. In the process of using the immunoaffinity column, the turbidity of the analytical sample solution often causes the clogging of the affinity column, which affects the antibody-antigen binding ability of the affinity column, and even subsequent steps such as washing and elution cannot be carried out normally.
[0007] The above factors limit the popularization and commercialization of immunoaffinity columns, and a simple and effective device is needed to solve these problems

Method used

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Examples

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Embodiment

[0033] Example: Application of immunoaffinity purification of tetrodotoxin in aquatic products.

[0034] Fill the tetrodotoxin antibody into the column cavity with a height of 1.2-1.3cm, cover the sieve plate, inject the active solution, tighten the upper and lower plugs, and store it at 0-5°C until use.

[0035] Accurately weigh 2.00 g of the fully homogenized horse-faced fish sample into a 50 mL stoppered centrifuge tube, add 10 mL of methanol solution containing 1% acetic acid, vortex for 2 min, ultrasonically extract in a 60°C water bath for 15 min, cool to room temperature, and 6000r / centrifuge for 5min. Pipette 5 mL of the supernatant to another 50 mL centrifuge tube, add 20 mL of PBS solution for dilution, adjust the pH to 7~8 with 1 mol / L sodium hydroxide solution, and wait for purification.

[0036] Take out the immunoaffinity column, remove the upper and lower plugs, release the active solution stored in the column, and load the sample solution. After loading the ...

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Abstract

The invention discloses an immunoaffinity method. The method comprises the following steps: filling an immunoaffinity column with a corresponding antibody matrix according to the concrete target of an analysis object; injecting an antibody active liquid to keep the activity of the antibody in order to make the highest liquid level be lower than the highest liquid level scale of the immunoaffinity column, screwing upper and lower end caps, and placing the obtained immunoaffinity column in appropriate ambient temperature; selecting single immunoaffinity column or combining a plurality of immunoaffinity columns according to the concentration range of the target, and determining the quantity of the immunoaffinity columns; and adding a target eluent, and collecting and analyzing an eluate flowing out of the immunoaffinity column(s). The immunoaffinity column(s) adopt(s) a telescopic spiral combination type immunoaffinity column. The immunoaffinity method improves the volume of the active liquid, enhances the integral sealing property of immunoaffinity, guarantees the sufficient antibody active liquid and prevents overflow and liquid leakage.

Description

technical field [0001] The present invention relates to an immunoaffinity method. Background technique [0002] As a sample pretreatment technology, solid phase extraction columns have been widely used in laboratories. With the continuous development of solid phase extraction technology, a new type of extraction column - immunoaffinity column is quietly replacing the traditional solid phase extraction column. Immunoaffinity column (IAC) is based on the antigen-antibody reaction, a separation and extraction column made of biomacromolecules with the characteristics of specific recognition and reversible binding to a class of biomacromolecules. It has unique selection specificity and good adsorption Purification performance. When the treated sample extract slowly passes through the immunoaffinity column, the analyte in the sample extract binds to the antibody in the column, while other impurities that are not bound are removed after washing, and then the column is washed with...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/60G01N33/53
Inventor 张小军严忠雍喻亮李佩佩方益祝银
Owner MARINE FISHERIES RES INST OF ZHEJIANG
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