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A kind of fumonisin immunoaffinity column and its preparation method and application

A fumonisin and immunophilic technology, which can be used in material testing products, measuring devices, instruments, etc., can solve the problems of affecting the efficiency of fumonisin purification by affinity purification columns, uncontrollable directionality, etc., saving time and cost, Simple operation and the effect of improving purification efficiency

Active Publication Date: 2016-06-22
SHANDONG MEIZHENG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since the coupling is non-specific, any part of the antibody may be coupled to the carrier, and the directionality is uncontrollable, which will inevitably affect the efficiency of the affinity purification column for purifying fumonisins

Method used

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  • A kind of fumonisin immunoaffinity column and its preparation method and application
  • A kind of fumonisin immunoaffinity column and its preparation method and application
  • A kind of fumonisin immunoaffinity column and its preparation method and application

Examples

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Effect test

Embodiment 1

[0042] Example 1: Preparation of Fumonisin Immunoaffinity Purification Column

Embodiment approach

[0043] A preferred embodiment of the present invention to prepare the fumonisin immunoaffinity purification column is as follows:

[0044] 1. Agarose Gel Activation

[0045] Take 2% agarose gel sepharose 2B and wash it with 20 times the volume of distilled water to wash away the remaining ethanol. Use a funnel to filter out the water. Weigh 5 grams of the wet gel after filtering off the water, add 7.5 ml of 0.8M NaOH, 2 ml of 30% epichlorohydrin, 2 mg / ml of sodium borohydride NaBH4, 5 ml, and shake the reaction at 25 ° C 8 hours.

[0046] After the reaction, wash with 50 ml of distilled water to remove the mixed epichlorohydrin in the gel.

[0047] 2. Coupling of Protein G to Activated Sepharose

[0048] Take 1 gram of activated agarose gel, and use coupling buffer (0.1M NaHCO 3 , 0.8M NaCl, pH8.9) and washed 3 times. Add 20ml of 2mg / ml protein G, and couple at room temperature for 4 hours.

[0049] The coupled agarose carrier was washed 3 times with 20 mM, pH 7.4 phosp...

Embodiment 2

[0068] Example 2: Purification and detection of fumonisins in corn samples using a fumonisin immunoaffinity column

[0069] In this example, normal corn samples were used to quantitatively add fumonisin standard substances, and then purified by immunoaffinity columns, and then detected by high performance liquid chromatography after purification to determine the recovery rate.

[0070] 1. Maize sample processing

[0071] 1) pulverize the corn sample;

[0072] 2) According to the standard of 15 micrograms per gram of sample, add fumonisin standard substance in the crushed corn sample;

[0073] 3) Weigh 1 sample and add 100mL of extract;

[0074] 4) High-speed homogenization for 2 minutes;

[0075] 5) filter with fast qualitative filter paper;

[0076] 6) Take 1.5mL filtrate, add 36mL diluent, shake and mix;

[0077] 7) filter with microfiber filter paper;

[0078] 8) Take all the filtrate for sample loading detection.

[0079] 2. Equilibrate the fumonisin immunoaffinity ...

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Abstract

The invention relates to a Fumonisin immunoaffinity column, and a making method and a use thereof. The immunoaffinity column is coupled to an agarose gel carrier by using a protein G, and an anti- Fumonisin antibody is used to couple with the protein G on agarose. The above obtained coupled Fumonisin antibody-protein G-agarose gel carrier composite carrier is filled to an affinity column. The immunoaffinity column is mainly used for purifying Fumonisin in foods, feeds, milk, blood samples and other various samples in order to provide convenience for later stage high performance liquid chromatography (HPLC) detection and fluorescence detection of Fumonisin in the samples.

Description

Technical field: [0001] The invention relates to a fumonisin immunoaffinity column, a preparation method and application thereof. It belongs to the field of food safety testing. Background technique: [0002] Fumonisins are a group of mycotoxins mainly produced by Fusarium moniliforme under certain temperature and humidity conditions. Fumonisins are widely distributed in nature and are very harmful, which have gradually attracted the attention of scholars at home and abroad. Currently, the discovered fumonisin structural analogues are mainly divided into four categories: A, B, C, and P. Studies have confirmed that fumonisins can cause equine leukomalacia (EL-EM), neurotoxicity and symptoms such as disturbance of consciousness, blindness and movement disorders, and even death in severe cases. Causes pulmonary edema syndrome (PPE) in pigs and can cause liver and esophagus damage. Fumonisins can also cause atherosclerotic changes in primates and induce liver cancer in rats....

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577
CPCG01N33/68
Inventor 柳家鹏张彦明孟丽敏徐家兰
Owner SHANDONG MEIZHENG BIOTECHNOLOGY CO LTD
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