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Fumonisin immunoaffinity column, and making method and use thereof

A fumonisin and immunophilic technology, applied in material testing products, measuring devices, instruments, etc., can solve the problems of uncontrollable directionality and affect efficiency, and achieve the effect of improving purification efficiency and improving capture ability.

Active Publication Date: 2014-12-03
山东美正生物科技有限公司
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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
It is bound to affect the efficiency of the affinity purification column for purifying fumonisins

Method used

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  • Fumonisin immunoaffinity column, and making method and use thereof
  • Fumonisin immunoaffinity column, and making method and use thereof
  • Fumonisin immunoaffinity column, and making method and use thereof

Examples

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Comparison scheme
Effect test

Embodiment 1

[0045] Example 1: Preparation of Fumonisin immunoaffinity purification column

Embodiment approach

[0046] A preferred embodiment of the present invention for preparing the fumonisin immunoaffinity purification column is as follows:

[0047] 1. Agarose gel activation

[0048] Take 2% agarose gel sepharose 2B and wash it with 20 times the volume of distilled water to wash away the remaining ethanol. Filter out the water with a funnel. Weigh 5 g of the wet gel after filtering 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 at 25°C React for 8 hours.

[0049] After the reaction, it was washed with 50 ml of distilled water to remove the epichlorohydrin mixed in the gel.

[0050] 2. Coupling of protein G and activated agarose gel

[0051] Take 1 gram of activated agarose gel and use coupling buffer (0.1M NaHCO 3 , 0.8M NaCl, PH8.9) wash 3 times. Add 20ml of 2mg / ml protein G and couple at room temperature for 4 hours.

[0052] The coupled agarose carrier was washed 3 times with 20 mM, pH 7.4 phosphate buffer PBS. Seph...

Embodiment 2

[0071] Example 2: Purification and detection of fumonisins in corn samples using fumonisin immunoaffinity column

[0072] In this example, a normal corn sample was used to quantitatively add fumonisins standard products, and then purified by an immunoaffinity column, and then detected by high performance liquid chromatography after purification. Determine the recovery rate.

[0073] 1. Corn sample processing

[0074] 1) Crush the corn sample;

[0075] 2) In accordance with the standard of 15 micrograms per gram of sample, add fumonisin standard to the crushed corn sample;

[0076] 3) Weigh 1 sample and add 100 mL of extract;

[0077] 4) High-speed homogenization for 2 minutes;

[0078] 5) Filter with fast qualitative filter paper;

[0079] 6) Take 1.5mL filtrate, add 36mL diluent, shake and mix;

[0080] 7) Filter with microfiber filter paper;

[0081] 8) Take all the filtrate for sample loading test.

[0082] 2. Equilibrate the fumonisin immunoaffinity purification column at room temperatur...

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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 an immunoaffinity column for fumonisins and a preparation method and application thereof. It belongs to the field of food safety testing. Background technique: [0002] Fumon isins are a group of mycotoxins mainly produced by the reproduction of Fusarium moniliforme under certain temperature and humidity conditions. Fumonisins are widely distributed in nature and have great harmfulness, and gradually attracted the attention of scholars at home and abroad. At present, the structural analogues of fumonisins that have been discovered are mainly divided into four categories: A, B, C, and P. Studies have confirmed that fumonisins can cause equine brain 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 atherosclerosis-...

Claims

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Application Information

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IPC IPC(8): G01N33/577
CPCG01N33/68
Inventor 柳家鹏张彦明孟丽敏徐家兰
Owner 山东美正生物科技有限公司
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