A dihydrocapsaicin polyclonal antibody immunoadsorbent, immunoaffinity column and its preparation method and application

A technology of dihydrocapsaicin and polyclonal antibodies, which is applied in the directions of solid adsorbent liquid separation, chemical instruments and methods, separation methods, etc. Complete removal of compounds and other problems to achieve the effect of low cost, high efficiency and simple operation

Active Publication Date: 2016-08-24
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Dihydrocapsaicin has the properties of fat solubility, good stability, and high boiling point, and the swill oil in the catering industry is one of the main sources of waste oil. It is difficult to completely remove compounds with the above properties in the current waste oil processing methods
At present, there is no relevant report on the immunoaffinity column of dihydrocapsaicin polyclonal antibody

Method used

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  • A dihydrocapsaicin polyclonal antibody immunoadsorbent, immunoaffinity column and its preparation method and application
  • A dihydrocapsaicin polyclonal antibody immunoadsorbent, immunoaffinity column and its preparation method and application
  • A dihydrocapsaicin polyclonal antibody immunoadsorbent, immunoaffinity column and its preparation method and application

Examples

Experimental program
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Effect test

Embodiment 1

[0036] Embodiment 1: the preparation of dihydrocapsaicin artificial antigen

[0037] Weigh 5.86g of N-(4-hydroxy-3-methoxybenzyl)nonanamide, 2.36g of maleic anhydride and 0.49g of N,N'-dicyclohexylcarbodiimide, and put 25mL of dichloromethane in React in the reaction bottle at room temperature for 0.5h, then place the above reaction solution in an ice bath to control the temperature within the range of 0-5°C, take 0.30g of 4-dimethylaminopyridine and 4mL of dichloromethane and place it in a constant pressure drop Mix well in the funnel, slowly add dropwise to the reaction bottle, and the dropwise addition is completed in 0.5h. After the dropwise addition is completed, keep the room temperature at 25°C and continue to stir for 6 hours, filter the reaction solution, and distill off the solvent under reduced pressure to obtain a light yellow oil. Thick product, then this crude product is passed through silica gel column chromatography, and column chromatography reagent: petroleum...

Embodiment 2

[0042] Example 2: Preparation of Dihydrocapsaicin Polyclonal Antibody

[0043] In this example, the dihydrocapsaicin artificial antigen obtained in Example 1 was used as the immunogen, and the dihydrocapsaicin antiserum was obtained by immunization, and the antiserum was obtained by two-step precipitation and purification with saturated ammonium sulfate, that is, the dihydrocapsaicin polyclonal antibody. The specific operation is as follows:

[0044] The dihydrocapsaicin artificial antigen was emulsified with Freund's complete adjuvant or Freund's incomplete adjuvant. The first immunization was carried out with Freund's complete adjuvant, and the subsequent immunizations were all emulsified with Freund's incomplete adjuvant. Balb / c mice were immunized with intramuscular injections, once every 3 weeks, and started from the third immunization, and the blood samples of Balb / c mice were centrifuged one week after each immunization to obtain anti- For serum, the titer of antiserum...

Embodiment 3

[0046] Example 3: Preparation of Dihydrocapsaicin Polyclonal Antibody Immunoaffinity Column Adsorbent and Immunoaffinity Column

[0047] The immunoaffinity column adsorbent of this embodiment comprises a solid phase carrier (polyacrylamide silica gel microspheres) and a dihydrocapsaicin polyclonal antibody coupled to the solid phase carrier, and the specific preparation method is as follows:

[0048] Weigh 1.0 g of polyacrylamide silica gel microspheres, put them into an Erlenmeyer flask, wash the microspheres alternately with pure water and pH=6 phosphate buffer; measure 5 mL of pH=6 phosphate buffer to dissolve the polyacrylamide Silica gel microspheres, stirred and mixed at room temperature 25°C for 10min, added dropwise 1mL of dihydrocapsaicin polyclonal antibody solution (concentration: 2mg / mL) with pH = 6, added 70mg of EDC, stirred at 4°C for 20-22h, A dihydrocapsaicin polyclonal antibody immunoadsorbent using polyacrylamide silica gel microspheres as a solid phase carr...

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Abstract

The invention relates to a dihydrocapsaicin polyclonal antibody immunoadsorbent, an immunoaffinity column, and a preparation method and application thereof. The immunoadsorbent comprises a solid carrier and a dihydrocapsaicin polyclonal antibody coupled on the solid phase carrier, wherein the dihydrocapsaicin polyclonal antibody is obtained by separating, purifying, then freezing and drying a dihydrocapsaicin artificial antigen immune animal obtained by coupling the dihydrocapsaicin artificial semiantigen (E)-4-(2-methoxy-4-(9 methylformamide) phenoxy base)-4-oxygen-2-crotonic acid which is described by formula I with bovine serum albumin (BSA). The immunoaffinity column disclosed by the invention can be specifically combined with dihydrocapsaicin, has a maximum column capacity of 220-250ng; and the subscript recovery rate can reach 83.1% and RSD (Relative Standard Deviation) is less than 6% by using methanol for elution. The immunoadsorbent can be used for preprocessing the sample of the dihydrocapsaicin content, and has the advantages of simplicity in operation, low cost, high efficiency and less consumption of organic reagents.

Description

Technical field: [0001] The invention relates to a dihydrocapsaicin polyclonal antibody immunoadsorbent, an immunoaffinity column, a preparation method and application thereof. Background technique [0002] In recent years, the act of mixing abnormal cooking oil (commonly known as gutter oil) into qualified cooking oil for profit has been repeatedly banned, which seriously threatens the health of consumers and has become an urgent problem in the field of food safety in my country. In my country's traditional dietary habits, pepper is a condiment with a large amount of use and a wide range of applications. Dihydrocapsaicin is the main chemical component that causes spicy taste, and it mainly exists in the fruits and seeds of Capsicum plants. Dihydrocapsaicin has the properties of fat solubility, good stability, and high boiling point, and the swill oil in the catering industry is one of the main sources of waste oil. It is difficult to completely remove compounds with the abo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01J20/24B01J20/281B01J20/30B01D15/38G01N33/543G01N33/531
CPCB01D15/3823B01J20/24B01J20/265B01J20/28021B01J20/28047G01N33/531G01N33/543
Inventor 李培武马飞张奇杨青青张良晓丁小霞
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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