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Universal immunoaffinity column for mycotoxin detection

A mycotoxin and general-purpose technology, which is applied in the field of general-purpose immunoaffinity columns, can solve the problems of small application range, low utilization rate of immunoaffinity columns, and lack of versatility, and achieve fast antibody coupling speed and improved The effect of processing efficiency and small coefficient of variation

Pending Publication Date: 2019-08-02
MEIZHENG BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the column tube is not versatile during use, and the utilization rate of the immunoaffinity column is low and the scope of use is small

Method used

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  • Universal immunoaffinity column for mycotoxin detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 An immunoaffinity column for the detection of aflatoxin B1

[0023] 1. Preparation of immunoaffinity column

[0024] (1) Sepharose 4B filler coupled with streptavidin

[0025] Place 1 mL of 1 mg / mL streptavidin in coupling buffer (0.1 mol / L NaHCO containing 0.5 mol / L NaCl at pH 8.3 3 ) in dialysis for 12 hours; swell the agarose dry powder Sepharose 4B activated with cyanogen bromide with 1mM hydrochloric acid, and then quickly wash it with coupling buffer in a sand core funnel to remove impurities; then add it to the streptavidin solution Add 10 g of activated Sepharose4B, mix well, and shake at room temperature for 1 hour; wash off uncoupled streptavidin with more than 5 times the volume of coupling buffer to obtain Sepharose4B-streptavidin coupling complex thing.

[0026] (2) Block active groups:

[0027] Transfer the Sepharose 4B filler modified with streptavidin into 5 times the volume of 0.1mol / L Tris-HCl buffer containing 0.5mol / L NaCl at pH 8.0 and...

Embodiment 2

[0054] Example 2 An immunoaffinity column for the detection of aflatoxin B1, deoxynivalenol, and zearalenone

[0055] 1. Preparation of immunoaffinity column

[0056] (1) Sepharose 4B filler coupled with streptavidin

[0057] Same as step (1) of Example 1.

[0058] (2) Block active groups:

[0059] Same as step (2) of Example 1.

[0060] (3) Washing:

[0061] Same as step (3) of Example 1.

[0062] (4) Column packing:

[0063] Same as step (4) of Example 1.

[0064] (5) Aflatoxin B1 antibody, deoxynivalenol antibody, zearalenone antibody conjugated biotin

[0065] Each antibody was individually coupled with biotin and freeze-dried separately, the steps were the same as step (5) of Example 1.

[0066] 2. Determination of actual sample recovery rate

[0067] Corn samples containing 5μg / kg, 10μg / kg, 50μg / kg aflatoxin B1, 145μg / kg, 877μg / kg, 1520μg / kg DON corn samples, 12.5μg / kg, 35.6μg / kg, Nine kinds of corn samples with 226.4 μg / kg zearalenone were pretreated, and then...

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Abstract

The invention provides a universal immunoaffinity column for detecting mycotoxin, which comprises a column tube, a sieve plate, a filler and a mycotoxin antibody, the bottom of the column tube is provided with a liquid outlet, the mycotoxin antibody is independently subpackaged in a storage tube, the mycotoxin antibody is coupled with biotin, and the filler is coupled with streptavidin. The fillerused by the immunoaffinity column is coupled with streptavidin, the filler only needs to be in affinity binding with an antibody modified by biotin to modify the antibody to the surface of the filler, and the filler is filled into a column tube to perform pretreatment on a sample. The column capacity of the prepared immunoaffinity column can reach 5000 ng; meanwhile, the recovery rate of actual sample detection is high, and the coefficient of variation is small, so that the immunoaffinity column has a good pretreatment effect and relatively high stability on samples, can simultaneously carryout high-flux adsorption on various targets, and has a wide application prospect in the field of food safety detection.

Description

technical field [0001] The invention belongs to the field of food safety detection, and in particular relates to a universal immunoaffinity column for mycotoxin detection. Background technique [0002] Mycotoxins are secondary metabolites produced by toxin-producing fungi under certain environmental conditions, which widely contaminate plant-derived products such as crops, food, and feed. The more common mycotoxins in grain, food and feed include: aflatoxin (AF), ochratoxin (ochratoxin, OT), fumonisin (fumonisin, FB), deoxynivalenol ( deoxynivalenol, also known as vomitoxin, DON), T-2 toxin, HT-2 toxin, patulin, etc. Mycotoxins can cause acute or chronic poisoning in humans and animals, and can damage the body's liver, kidneys, nerve tissue, hematopoietic tissue, and skin tissue. In fact, the toxic effects of many mycotoxins are multi-organ, that is, damage Two or more organs and tissues. According to the estimates of the Food and Agriculture Organization of the United Na...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543
CPCG01N33/543
Inventor 张勋
Owner MEIZHENG BIO TECH CO LTD
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