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Tetrodotoxin immunoaffinity column and making method thereof

A technology of tetrodotoxin and immunophile, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., to achieve the effect of improving purification efficiency, easy operation, and improving antibody binding ability

Inactive Publication Date: 2016-04-06
MEIZHENG BIO TECH 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
It is bound to affect the efficiency of the affinity purification column for purifying tetrodotoxin

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0038] Example: Preparation of Tetrodotoxin Immunoaffinity Purification Column

Embodiment approach

[0039] A preferred embodiment of the present invention to prepare tetrodotoxin immunoaffinity purification column is as follows:

[0040] 1. Agarose Gel Activation

[0041] Take 2% agarose gel sepharose 2B and wash it with 20 times the volume of distilled water to remove the remaining ethanol. Use a funnel to filter out the water. Weigh 5 grams of the wet gel after filtering out the water, add 7.5 ml of 0.8M NaOH, 2 ml of 30% epichlorohydrin, 2 mg / ml of sodium borohydride NaBH4, 5 ml, shake the table at 25 ° C React for 8 hours.

[0042] After the reaction, wash with 50 ml of distilled water to remove the mixed epichlorohydrin in the gel.

[0043] 2. Coupling of Protein G to Activated Sepharose

[0044] Take 1 g of the activated agarose gel and wash with 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.

[0045] The coupled agarose carrier was washed 3 times with 20 mM, pH 7.4 ...

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PUM

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Abstract

The invention relates to a tetrodotoxin immunoaffinity column and a making method thereof, and belongs to the field of detection of toxins in aquatic products. The above immunoaffinity purification column is coupled to an agarose gel carrier by using a protein G, and an anti-tetrodotoxin antibody is coupled to the protein G on agarose. A cross-linking agent is used to cross-link the above obtained anti-tetrodotoxin antibody-protein G-agarose gel carrier, and then the affinity column is filled. The immunoaffinity column is mainly used for purifying tetrodotoxin in the aquatic products.

Description

Technical field: [0001] The invention relates to an immunoaffinity column for tetrodotoxin and a preparation method thereof, belonging to the field of aquatic toxin detection. Background technique: [0002] Tetrodotoxin (TTX) is an alkaloid (aminoperhydroquinazole) contained in porpoise fish (commonly known as puffer fish), other marine organisms, and salamanders, salamanders, frogs, toads, flatworms and other animals. phenotype compound), is one of the most toxic neurotoxins found in nature, and its molecular formula is C 11 h 17 o 8 N 3 , with a molecular weight of 319, is a small molecular weight, non-protein neurotoxin, its toxicity is more than 1250 times higher than that of highly toxic sodium cyanide, and 0.5mg can cause death. Tetrodotoxin has a local stimulating effect on the intestinal tract, and quickly acts on nerve endings and nerve centers after absorption, hindering nerve conduction, thereby causing nerve paralysis and death. Its specific mechanism of a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/569G01N33/531
Inventor 柳家鹏
Owner MEIZHENG BIO TECH CO LTD
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