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Preparation method of deoxynivalenol immunoaffinity column

A technology of vomitoxin and immunoaffinity, which is applied in the field of immunoaffinity column preparation, can solve the problems of high sample purity requirements, interference, lack of selectivity, etc., and achieve the effect of reducing the probability of vomitoxin

Inactive Publication Date: 2014-05-21
JIANGSU WISE SCI & TECH DEV
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  • Claims
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Problems solved by technology

Instrumental method detection has the advantages of high sensitivity and strong specificity, but it requires high purity of the sample and needs to go through a complicated purification process. Traditional sample pretreatment techniques, such as liquid-liquid extraction, solid-phase extraction, and solid-phase microextraction etc., the steps are cumbersome, lack of selectivity, easily interfered by other components, and low accuracy
Moreover, the toxicity of vomitoxin is severe, and the operator is repeatedly exposed to the environment of vomitoxin during the pretreatment process, which increases the risk of the experiment

Method used

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  • Preparation method of deoxynivalenol immunoaffinity column
  • Preparation method of deoxynivalenol immunoaffinity column

Examples

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Embodiment 1

[0019] Example 1 Preparation of Deoxynivalenol Immunoaffinity Column

[0020] (1) Coupling

[0021] Take 10 mg of the monoclonal antibody against vomitoxin purified by caprylic acid-ammonium sulfate method, dialyze in 0.1 mol / L NaHCO3 solution overnight, and adjust the concentration to 2 mg / mL;

[0022] Weigh 800mg of CNBr-Sepharose 4B dry gel, fully swell in 10mL 1.0 mmol / L HCL; wash 3 times with 0.1 mol / L NaHCO3 solution which is more than 10 times the volume of wet gel, and centrifuge at 5000rpm for 1min;

[0023] Mix the wet glue and antibody thoroughly, and stir overnight at room temperature;

[0024] Wash with 50mL 0.1 mol / L NaHCO3 solution, collect the washing liquid, identify it by ultraviolet light, and calculate the coupling rate. The formula for calculating the coupling rate is:

[0025]

[0026] The test results of the coupling rate showed that the coupling rate between the DON monoclonal antibody and CNBr-Sepharose 4B was 92.45%.

[0027] (2) closed

[002...

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Abstract

The invention discloses a preparation method of an immunoaffinity column based on deoxynivalenol monoclonal antibodies. An affinity adsorbent obtained through coupling of monoclonal antibodies obtained through cell fusion and cyanogen bromide activated agarose 4B is filled in a solid phase extraction tube to obtain the immunoaffinity column capable of specific adsorption of deoxynivalenol. The immunoaffinity column prepared by the invention can specifically combine with deoxynivalenol, has a maximal binding capacity of about 240ngDON, and reaches recovery rate of no less than 90% for three times of repeated usage. The invention can be used in the pretreatment method of detection of deoxynivalenol residues in cereals and cereal products.

Description

technical field [0001] The invention belongs to the technical field of detection, and in particular relates to a preparation method of an immunoaffinity column for vomitoxin. Background technique [0002] Deoxynivalenol (DON), also known as deoxynivalenol (DON), is a toxin that can cause vomiting and was isolated from corn contaminated by Fusarium graminearum in 1970. It is mainly produced by Fusarium graminearum ( Fusarium graminearum ) produced, belonging to Trichothecenes refining family compound type B. It is relatively stable to heat, and its toxicity cannot be destroyed by general cooking and heating. It is sensitive to alkaline environment. Washing grain with sodium carbonate solution can remove about 70% of DON. [0003] DON poisoning animals manifested as vomiting, loss of appetite, weight loss, miscarriage, weak offspring and stillbirth, and increased mortality. After human DON poisoning, nausea, vomiting, stomach discomfort, abdominal pain, diarrhea, headache, d...

Claims

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Application Information

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IPC IPC(8): B01D15/38B01J20/281G01N1/34
Inventor 杜道林曾昆
Owner JIANGSU WISE SCI & TECH DEV
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