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Immunoadsorbent and composite affinity column for purification of fumonisin B1, aflatoxin, ochratoxin A and zearalenone

A technology of zearalenone and aflatoxin, which is applied in the direction of solid adsorbent liquid separation, chemical instruments and methods, antifungal/algae/lichen immunoglobulin, etc., to achieve the effect of stable performance

Active Publication Date: 2017-07-25
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

[0007] In order to solve the detection of fumonisin B in the prior art 1 , aflatoxin, ochratoxin A, and zearalenone urgently need a stable immunoaffinity column. The present invention provides a method for purifying fumonisin B 1 , aflatoxin, ochratoxin A, zearalenone composite affinity column and its preparation method and application

Method used

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  • Immunoadsorbent and composite affinity column for purification of fumonisin B1, aflatoxin, ochratoxin A and zearalenone
  • Immunoadsorbent and composite affinity column for purification of fumonisin B1, aflatoxin, ochratoxin A and zearalenone
  • Immunoadsorbent and composite affinity column for purification of fumonisin B1, aflatoxin, ochratoxin A and zearalenone

Examples

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Effect test

Embodiment 1

[0042] The acquisition of embodiment 1 aflatoxin universal monoclonal antibody

[0043] The universal monoclonal antibody to aflatoxin is secreted and produced by the hybridoma cell line 1C11 with the deposit number CCTCC NO.C201013. Specifically, it is pre-prepared according to the method reported in the patent with the authorization number ZL 201010245095.5. The preparation method is as follows: the obtained hybridoma The cell line 1C11 was injected into BALB / c mice treated with Freund's incomplete adjuvant in advance, and the ascites of the mice was collected and purified to obtain aflatoxin universal B monoclonal antibody. Among them, the purification method is the octanoic acid-ammonium sulfate method, and the specific operation is: filter the mouse ascites with double-layer filter paper, centrifuge the filtered ascites at 4°C, 12000r / min for more than 15min, absorb the supernatant, and dilute the supernatant with 4 times the volume Mix acetate buffer solution, slowly add...

Embodiment 2

[0045] Embodiment 2 Obtaining of ochratoxin A monoclonal antibody;

[0046] Ochratoxin A monoclonal antibody is secreted and produced by the hybridoma cell line 1H2 with the deposit number CCTCC NO.C201329. Specifically, it is prepared in advance according to the method reported in the patent application number 201310115921.8. The preparation method is: hybridoma cell line 1H2 Inject into the abdomen of BALB / c mice treated with Freund's incomplete adjuvant in advance, collect the ascites of the mice, and purify to obtain ochratoxin A monoclonal antibody. The purification method is caprylic acid-ammonium sulfate method, and the specific steps are: filter mouse ascites with double-layer filter paper, centrifuge at 12,000 r / min for more than 15 minutes at 4°C, absorb the supernatant, and mix the obtained ascites supernatant with 4 times the volume of vinegar Mix salt buffer solution, add n-octanoic acid slowly under stirring, the volume of n-octanoic acid required per ml of ascit...

Embodiment 3

[0048] Example 3 Obtaining of Zearalenone Monoclonal Antibody

[0049] The zearalenone monoclonal antibody is secreted and produced by the hybridoma cell line 2D3 with the deposit number CCTCC NO.C201328. Specifically, it is prepared in advance according to the method reported in the patent application number 201310115825.3. The preparation method is as follows: the hybridoma cell line 2D3 was injected into the abdomen of BALB / c mice pre-treated with Freund's incomplete adjuvant, the ascites of the mice was collected, and purified to obtain the zearalenone monoclonal antibody; the purification method is caprylic acid-ammonium sulfate The specific steps are as follows: filter mouse ascites with double-layer filter paper, centrifuge at 12,000 r / min for more than 15 minutes at 4°C, absorb the supernatant, mix the obtained ascites supernatant with 4 times the volume of acetate buffer, and slowly add n-octanoic acid, the volume of n-octanoic acid required per milliliter of ascites ...

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Abstract

The present invention relates to an immunoadsorbent and a composite affinity column for purification of fumonisin B1, aflatoxin, ochratoxin A and zearalenone. The immunoadsorbent comprises a solid carrier and fumonisin B1, aflatoxin, ochratoxin A and zearalenone monoclonal antibodies coupled with the solid carrier, the fumonisin B1 monoclonal antibody is secreted from hybridoma cell line Fm7A11 with the preservation number of CCTCCNO.C201636, the hybridoma cell line Fm7A11 is preserved in CCTCC in March 29, 2016 in Wuhan University Wuhan China. The composite affinity column is loaded with the immunoadsorbent with the fumonisin B1, aflatoxin, ochratoxin A and zearalenone monoclonal antibodies, and is used for purification of the fumonisin B1, aflatoxin, ochratoxin A and zearalenone. The composite affinity column can be used for purification and detection of fumonisin B1, aflatoxin, ochratoxin A and zearalenone toxin samples.

Description

technical field [0001] The invention relates to a purifying fumonisin B1, aflatoxin, ochratoxin A, zearalenone immune adsorbent and a composite affinity column. Background technique [0002] Fumonisins are water-soluble metabolites produced by Fusarium spp. under certain temperature and humidity conditions. Fumonisins can interfere with the normal physiological functions of plants in a low concentration range. They are non-enzyme compounds that have toxic effects on plant metabolism, and belong to mycotoxins and non-host-specific toxins. It is mainly distributed on crops mainly including corn, sorghum and wheat, and can cause agricultural economic losses such as seedling withering, root, stem and seed rot. Fumonisins can cause various specific toxicological effects on livestock and poultry and experimental animals, such as leukomalacia in horses and rabbits, which is manifested as: neurotoxicity, disturbance of consciousness, blindness and ataxia and other symptoms, severe ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/14G01N33/577G01N30/02B01J20/286B01D15/22B01D15/20
CPCB01D15/20B01D15/22B01J20/286C07K16/14G01N30/02G01N33/577
Inventor 李培武胡小风张兆威张奇张文唐晓倩
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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